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GOOD MORNING.

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IT'S FANTASTIC TO SEE SO MANY

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PEOPLE IN PERSON.

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I DON'T THINK THIS ROOM HAS BEEN

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THIS FULL IN PROBABLY SINCE

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PRE-COVID.

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SO FOR THOSE WHO DON'T KNOW ME

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OR MIGHT NOT RECOGNIZE ME IN THE

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FANCY CLOTHES, I'M ERIC FREED,

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DIRECTOR OF THE DYNAMICS AND

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REPLICATION PROGRAM, ON BEHALF

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OF THE STAFF AND FACULTY OF THE

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CRP I'D LIKE TO WELCOME YOU TO

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THE CELEBRATION OF ALAN REIN'S

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CAREER.

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HIGHLIGHTS TO GET US STARTED,

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ALAN GRADUATED FROM REID COLLEGE

18
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IN 1961, Ph.D. RESEARCH WITH

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HARRY RUBIN AT UC-BERKELEY

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BEFORE DOING POSTDOCTORAL

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RESEARCH AT M.I.T., WITH SHELDON

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PENMAN.

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IN THE 70s HE WORKED AT LITTON

24
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BIOMEDICS THAT DID

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GOVERNMENT-FUNDED CONTRACT

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RESEARCH.

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LATE 70s WORKED IN TUMOR VIRUS

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GENETICS AT NCI BETHESDA.

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IN 1979, ALAN MOVED TO FREDERICK

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TO SERVE AS A SENIOR RESEARCH

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SCIENTIST, MICROBIOLOGY OF

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RETROVIRUS SECTION, IN THE

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CANCER RESEARCH FACILITY.

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BETWEEN 1984 AND 1999 ALAN WAS

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HEAD OF THE RETROVIRAL GENETICS

36
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SECTION AT ABL IN FREDERICK.

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AND SINCE 1999, ALAN HAS BEEN

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HEAD OF THE RETROVIRUS ASSEMBLY

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SECTION IN THE HIV DRP.

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HE HAS MANY HONORS AND

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ACCOMPLISHMENTS OVER THE YEARS,

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SERVED ON EDITORIAL BOARDS OF

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SEVERAL JOURNALS INCLUDING JBC,

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VIROLOGY, IN 2012 RECEIVED THE

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NIH DIRECTOR'S AWARD, 2014

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ELECTED FELLOW AMERICAN ACADEMY

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OF MICROBIOLOGY.

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I WOULD LIKE TO SAY A FEW WORDS

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ABOUT HIS GROUNDBREAKING

50
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CONTRIBUTIONS TO THE FIELD OF

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RETROVIROLOGY.

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I COULD SPEND HALF THE DAY DOING

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THIS.

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CONDENSING THINGS DOWN -- HELLO?

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PLEASE MUTE YOURSELF.

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>> WAS THAT YOU TALKING?

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>> NO, THE CHAIR OF THE
MEETING.

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>> PLEASE MUTE YOURSELF.

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EARLY TO MID-1970s, ALAN DID

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GROUNDBREAKING RESEARCH ON Fv1

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REVICES OF MLV INFECTION,

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JONATHAN STOYE WILL SAY MORE

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LATER.

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EARLY INDICATIONS MEMBRANE

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BINDING OF GAG IS ESSENTIAL FOR

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VIRUS ASSEMBLY SUBSEQUENTLY

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SHOWN FOR HIV AND MANY OTHER

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RETROVIRUSES.

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IN 1988, ALAN SHOWED POINT

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MUTANTS FAILED TO PACKAGE VIRAL

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RNA, PROVIDING EVIDENCE FOR

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SPECIFIC RNA RECOGNITION BY THE

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ZINC FINGER-LIKE DOMAIN, NC ZINC

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FINGER DOMAIN WE'RE FAMILIAR

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WITH.

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1994 SHOWED THE MLB

77
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TRANSMEMBRANE GLYCOPROTEIN IS

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CLEAVED TO ACTIVATE FUSION

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ACTIVITY, THE FIRST INDICATION

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OF A CONNECTION BETWEEN

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ACTIVATION AND VIRUS MATURATION.

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IN 2001, ALAN SHOWED RNA IS A

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STRUCTURAL ELEMENT IN RETROVIRUS

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PARTICLES, IN 2001 SHOWED THE

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MODULATION OF HIV-LIKE PARTICLE

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ASSEMBLY IN IN VITRO PREDATED BY

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NEARLY TWO DECADES THE CURRENT

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APPRECIATION FOR THE ROLE OF IP6

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IN RETROVIRAL ASSEMBLY AND

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MATURATION.

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TOGETHER WITH NCI FREDERICK

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CONTRIBUTED TO SOLVING THE

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STRUCTURE OF THE HIV REN

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RESPONSE ELEMENT AND DISSECTED

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SPECIFIC BINDING OF HIV-1 GAG TO

96
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PACKAGING SIGNAL IN VIRAL RNA,

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GAG BINDING SPECIFICALLY TO

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VIRAL RNA IS TRIGGER FOR VIRUS

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ASSEMBLY EXPLAINING SPECIFICITY

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OF RNA PACKAGING.

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SO, IN ADDITION TO BEING

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ENORMOUS CONTRIBUTOR TO THE

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FIELD OF RETROVIROLOGY, ALAN WAS

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THEN ATTENDING FOR MANY YEARS,

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MANY CONFERENCES IN THE FIELD,

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INCLUDING THE COLD SPRING HARBOR

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RETROVIRUS MEETING.

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HERE IS ALAN IN 1989.

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AND WITH JACKIE BENTLEY, HERE

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WITH US TODAY, IN 1991. 

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HERE IS ALAN IN 1998 WITH JOHN,

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IN THE FRONT ROW, JUST LIKE AT

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COLD SPRING HARBOR.

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ALAN AND DELPHINE IN 1999.

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THEY ACTUALLY LOOK EXACTLY THE

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SAME NOW AS THEY DID THEN.

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IT'S REALLY REMARKABLE. 

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IN 2001 WITH STEVE.

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SO I'LL PROVIDE A FEW QUICK

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HIGHLIGHTS OF THE PROGRAM WE

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HAVE A LINEUP OF DISTINGUISHED

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SPEAKERS TODAY.

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BEGINNING WITH FRANCOIS

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BARRE-SINOUSSI FROM INSTITUT

125
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PASTEUR, JONATHAN STOYE.

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AFTER THE BREAK DELPHINE MURIAUX

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AND MAURICIO COMAS-GARCIA, AND

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WE'LL HEAR ON TRANSCRIPTION

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START HETEROGENEITY; AND THE TWO

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FORMERS DIRECTORS, VIRTUAL

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PRESENTATION FROM PETER AND

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FINALLY ALAN REIN WILL PROVIDE

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CLOSING REMARKS.

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THERE WILL BE AN AFTER PARTY

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CELEBRATION AT THE END OF THE

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DAY AT STEINHARDT BREWERY,

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DOWNTOWN FREDERICK.

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FINALLY, ALAN, I CONGRATULATE

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YOU ON YOUR GROUNDBREAKING

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00:06:39,799 --> 00:06:41,568
CONTRIBUTIONS TO THE FIELD OF

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00:06:41,568 --> 00:06:42,769
RETROVIROLOGY, AND WISH YOU THE

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BEST FOR A LONG AND HAPPY

143
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RETIREMENT.

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ALAN WILL CONVERT TO EMERITUS

145
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STATUS AT THE END OF THE YEAR SO

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WE HOPE YOU'LL STAY ENGAGED IN

147
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DRP ACTIVITIES.

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HOWEVER, WE HOPE HE WILL ALSO

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SPEND TIME WITH FRIENDS AND

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FAMILY, OLD AND NEW HOBBIES AND

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INTERESTS.

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YOU KNOW HE'S AN AVID BIRD

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WATCHER, FOR EXAMPLE.

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SO WITHOUT FURTHER DELAY I WILL

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WELCOME ALAN TO THE PODIUM TO

156
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KICK OFF TODAY'S PROGRAM.

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>> THANK YOU, ERIC.

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THAT WAS MORE THAN I COULD WISH

159
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FOR.

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SO, I ASKED TO INTRODUCE EACH

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SPEAKER MYSELF, AND JUST SAY A

162
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FEW WORDS, NOT ABOUT THE USUAL

163
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THEY HAVE SO MANY AWARDS, BLAH,

164
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BLAH, BLAH, BUT WHAT THEY ARE

165
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DOING HERE, WHAT IS THE

166
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CONNECTION SUCH THAT THEY ARE

167
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PARTICIPATING IN THIS EVENT.

168
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SO, FIRST SPEAKER REALLY NEEDS

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NO INTRODUCTION, FRANCOISSE IS

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THE PERSON WHO ISOLATED HIV, AND

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BEFORE SHE ISOLATED HIV, IN FACT

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BEFORE HIV APPEARED, SHE STUDIED

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MURINE VIRUSES IN THE LAB OF BOB

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BASSEN, IN BETHESDA, AS DID I.

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AND SO WE WORKED SIDE BY SIDE IN

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THE 19, UM, '70s, MID-'70s,

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LATE 70s.

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WE'VE BEEN FRIENDS EVER SINCE.

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AND SO IT'S A GREAT PLEASURE TO

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SAY HELLO AGAIN TO MY OLD FRIEND

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FRANCOISE.

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>> THANK YOU VERY MUCH.

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GOOD MORNING, EVERYONE.

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TODAY WE GATHER TO HONOR OUR

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COLLEAGUE AND DEAR FRIEND ALAN,

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PROFESSIONAL LIFE FOR ADVANCES

187
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IN SCIENCE ON RETROVIRUSES.

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I'M FILLED WITH BOTH JOY AND

189
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NOSTALGIA, WITH GREAT MEMORIES

190
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COMING BACK IN MY MIND.

191
00:09:09,849 --> 00:09:16,623
AS IT WAS SAID BY ERIC, Ph.D.

192
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IN BERKELEY, M.I.T., ALAN IS

193
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ENJOYING THE NATIONAL CANCER

194
00:09:26,032 --> 00:09:28,134
INSTITUTE IN THE '70s, FIRST

195
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IN BETHESDA, AND THEN IN

196
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FREDERICK.

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THAT IS WHEN OUR PATHS FIRST

198
00:09:36,443 --> 00:09:42,582
CROSSED IN THE NCI LAB AT

199
00:09:42,582 --> 00:09:43,683
BIONETTICS IN BETHESDA.

200
00:09:43,683 --> 00:09:51,491
AT THAT TIME, I WAS THERE TO

201
00:09:51,491 --> 00:09:52,225
SUCCEED SYLVIE, WHO

202
00:09:52,225 --> 00:09:55,628
UNFORTUNATELY CANNOT BE WITH US

203
00:09:55,628 --> 00:09:55,929
TODAY.

204
00:09:55,929 --> 00:10:02,535
I WAS THERE FOR PART OF

205
00:10:02,535 --> 00:10:04,070
RESTRICTION OF MURINE VIRUS IN

206
00:10:04,070 --> 00:10:08,007
THE LABORATORY OF BOB BASSEN, AS

207
00:10:08,007 --> 00:10:09,209
WAS MENTIONED, UNFORTUNATELY BOB

208
00:10:09,209 --> 00:10:14,481
PASSED AWAY A YEAR AGO.

209
00:10:14,481 --> 00:10:18,852
I STILL VIVIDLY REMEMBER THAT

210
00:10:18,852 --> 00:10:21,588
PERIOD WITH YOU, ALAN, BOB,

211
00:10:21,588 --> 00:10:24,457
BRENDA, AND OTHERS.

212
00:10:24,457 --> 00:10:27,961
I WAS YOUNG, EAGER, VERY EXCITED

213
00:10:27,961 --> 00:10:29,996
TO LEARN ABOUT RETROVIRUSES FROM

214
00:10:29,996 --> 00:10:34,667
YOU AND PERHAPS A LITTLE NERVOUS

215
00:10:34,667 --> 00:10:37,137
ABOUT BEING OUT OF THE COUNTRY

216
00:10:37,137 --> 00:10:41,407
WITH MY POOR ENGLISH. 

217
00:10:41,407 --> 00:10:49,849
BUT I VERY RAPIDLY SENSED THE

218
00:10:49,849 --> 00:10:52,018
INTELLECT ENVIRONMENT, I NOT

219
00:10:52,018 --> 00:10:53,653
ONLY FOUND FANTASTIC MENTORS

220
00:10:53,653 --> 00:11:00,660
WITH BOB AND YOU, BUT ALSO

221
00:11:00,660 --> 00:11:01,761
LIFELONG FRIENDS.

222
00:11:01,761 --> 00:11:04,264
BELIEVE IT OR NOT, ALAN, WE HAVE

223
00:11:04,264 --> 00:11:09,202
KNOWN EACH OTHER FOR NEARLY HALF

224
00:11:09,202 --> 00:11:10,303
A CENTURY.

225
00:11:10,303 --> 00:11:13,239
ALMOST 15 YEARS AGO ALREADY YOU

226
00:11:13,239 --> 00:11:16,976
WERE ATTENDING IN PERSON WITH

227
00:11:16,976 --> 00:11:22,448
BOB AND DAN, ANOTHER LIFELONG

228
00:11:22,448 --> 00:11:24,751
NCI FRIEND, AT MEETING ORGANIZED

229
00:11:24,751 --> 00:11:27,754
FOR MY RETIREMENT AT PASTEUR

230
00:11:27,754 --> 00:11:30,023
INSTITUTE IN OCTOBER 2015.

231
00:11:30,023 --> 00:11:32,525
SO OBVIOUSLY I WANTED TO BE

232
00:11:32,525 --> 00:11:36,563
THERE TODAY FOR YOU, AT LEAST AT

233
00:11:36,563 --> 00:11:37,463
DISTANCE.

234
00:11:37,463 --> 00:11:43,136
LET ME TAKE A MOMENT TO BRIEFLY

235
00:11:43,136 --> 00:11:46,439
REFLECT ON NOT ONLY YOUR

236
00:11:46,439 --> 00:11:47,240
SCIENTIFIC ACHIEVEMENTS, THAT

237
00:11:47,240 --> 00:11:50,810
WAS ALREADY -- THEY WERE ALREADY

238
00:11:50,810 --> 00:11:54,314
MENTIONED, BY ERIC, BUT ALSO THE

239
00:11:54,314 --> 00:11:56,049
INCREDIBLE PERSON YOU ARE.

240
00:11:56,049 --> 00:12:00,219
AS YOU ALL KNOW HERE, ALAN'S

241
00:12:00,219 --> 00:12:03,957
PASSION AND COMMITMENT TO

242
00:12:03,957 --> 00:12:07,760
SCIENCE IS NOTHING SHORT OF

243
00:12:07,760 --> 00:12:08,094
INSPIRING.

244
00:12:08,094 --> 00:12:09,629
HE HAS MADE SIGNIFICANT ADVANCES

245
00:12:09,629 --> 00:12:12,031
THAT WERE MENTIONED TO OUR

246
00:12:12,031 --> 00:12:14,667
UNDERSTANDING OF SEVERAL ASPECTS

247
00:12:14,667 --> 00:12:19,138
OF REPLICATION OF MURINE AND

248
00:12:19,138 --> 00:12:20,907
HUMAN RETROVIRUSES INCLUDING

249
00:12:20,907 --> 00:12:21,874
HIV.

250
00:12:21,874 --> 00:12:24,410
ACCORDING TO THE AGENDA OF THE

251
00:12:24,410 --> 00:12:27,680
MEETING TODAY, THE NEXT SPEAKER

252
00:12:27,680 --> 00:12:30,516
WILL CERTAINLY MENTION ALAN'S

253
00:12:30,516 --> 00:12:34,887
CONTRIBUTION TO OUR KNOWLEDGE OF

254
00:12:34,887 --> 00:12:37,290
MOLECULAR, STRUCTURING, AND

255
00:12:37,290 --> 00:12:40,460
FUNCTIONAL MECHANISMS INVOLVED

256
00:12:40,460 --> 00:12:42,328
IN VIRAL ACTIVITY AND

257
00:12:42,328 --> 00:12:50,103
MATURATION, REGULATION OF

258
00:12:50,103 --> 00:12:50,536
INFECTION.

259
00:12:50,536 --> 00:12:58,211
BUT PIONEERING WORK ON Fv1, AS

260
00:12:58,211 --> 00:13:01,481
WAS JUST MENTIONED, AND ON THE

261
00:13:01,481 --> 00:13:07,053
INTERACTION OF HIV-1 GAG WITH

262
00:13:07,053 --> 00:13:08,588
NUCLEIC ACID LEADING TO

263
00:13:08,588 --> 00:13:10,123
CONFIRMATIONAL CHANGES REQUIRED

264
00:13:10,123 --> 00:13:11,658
FOR VIRAL ASSEMBLY.

265
00:13:11,658 --> 00:13:14,961
ALAN'S RESEARCH HAS NOT ONLY

266
00:13:14,961 --> 00:13:17,263
ADVANCED OUR KNOWLEDGE BUT IT

267
00:13:17,263 --> 00:13:21,834
HAS ALSO PAVED THE WAY FOR

268
00:13:21,834 --> 00:13:25,905
FUTURE DISCOVERIES IN THE FIELD.

269
00:13:25,905 --> 00:13:30,376
BUT BEYOND HIS IMPRESSIVE

270
00:13:30,376 --> 00:13:31,244
SCIENTIFIC CONTRIBUTIONS WHAT

271
00:13:31,244 --> 00:13:34,414
SETS ALAN APART IS HIS

272
00:13:34,414 --> 00:13:34,981
PERSONALITY.

273
00:13:34,981 --> 00:13:43,289
HE'S A MAN OF GENEROSITY, ALWAYS

274
00:13:43,289 --> 00:13:44,290
STANDING BY HIS PRINCIPLES AND

275
00:13:44,290 --> 00:13:46,592
HUMAN VALUES.

276
00:13:46,592 --> 00:13:50,630
IN A WORLD THAT OFTEN

277
00:13:50,630 --> 00:13:54,801
PRIORITIZES ACCOLADES, ALAN HAS

278
00:13:54,801 --> 00:13:56,769
ALWAYS REMAINED REFRESHINGLY

279
00:13:56,769 --> 00:13:57,203
MODEST.

280
00:13:57,203 --> 00:14:01,674
HE HAS NEVER SOUGHT THE

281
00:14:01,674 --> 00:14:05,411
SPOTLIGHT BUT RATHER SHINING IT

282
00:14:05,411 --> 00:14:08,381
ON OTHERS, MENTORING COUNTLESS

283
00:14:08,381 --> 00:14:10,683
STUDENTS AND COLLEAGUES ALONG

284
00:14:10,683 --> 00:14:12,752
THE WAY.

285
00:14:12,752 --> 00:14:16,556
HIS LOYALTY AND FRIENDSHIP HAVE

286
00:14:16,556 --> 00:14:17,757
BEEN UNWAVERING.

287
00:14:17,757 --> 00:14:21,627
THROUGH THE HIGHS AND LOWS OF

288
00:14:21,627 --> 00:14:24,797
OUR CAREERS, ALAN WAS ALWAYS

289
00:14:24,797 --> 00:14:27,200
THERE, OFFERING OR LISTENING

290
00:14:27,200 --> 00:14:30,036
HERE AND FRIENDLY SUPPORT.

291
00:14:30,036 --> 00:14:33,673
HE HAS ALWAYS REMINDED US THAT

292
00:14:33,673 --> 00:14:39,145
SCIENCE IS NOT JUST ABOUT DATA,

293
00:14:39,145 --> 00:14:41,681
REPLICATION, BUT ABOUT THE

294
00:14:41,681 --> 00:14:45,051
PEOPLE WE WORK AND FOR AND ABOUT

295
00:14:45,051 --> 00:14:49,989
THE RELATIONSHIP WE BUILD, ABOUT

296
00:14:49,989 --> 00:14:51,524
COLLABORATION, ABOUT SHARING

297
00:14:51,524 --> 00:14:52,525
GOALS.

298
00:14:52,525 --> 00:14:54,961
I HAVE WONDERFUL MEMORIES OF

299
00:14:54,961 --> 00:14:57,463
TIME WE SPENT TOGETHER IN THE

300
00:14:57,463 --> 00:15:02,402
LAB, AT HOME, OR SOMEWHERE ELSE

301
00:15:02,402 --> 00:15:04,704
EXCHANGING EXPERIENCE, SHARING

302
00:15:04,704 --> 00:15:07,306
OUR YEARS, AND DISCUSSING NOT

303
00:15:07,306 --> 00:15:11,911
ONLY SCIENCE BUT ALSO LIFE,

304
00:15:11,911 --> 00:15:14,547
DREAMS, AND ASPIRATIONS.

305
00:15:14,547 --> 00:15:19,385
ALAN, YOU BELIEVED IN ME WHEN I

306
00:15:19,385 --> 00:15:23,089
SOMETIMES DID NOT BELIEVE IN

307
00:15:23,089 --> 00:15:24,323
MYSELF, OFFERING YOUR GUIDANCE,

308
00:15:24,323 --> 00:15:28,327
ENCOURAGING ME TO PURSUE MY

309
00:15:28,327 --> 00:15:28,761
JOURNEY.

310
00:15:28,761 --> 00:15:31,430
YOU CERTAINLY DID IT FOR MANY

311
00:15:31,430 --> 00:15:33,833
OTHERS AROUND YOU.

312
00:15:33,833 --> 00:15:37,003
YOUR ABILITY TO MENTOR IS A GIFT

313
00:15:37,003 --> 00:15:41,040
THAT HAS BEEN SO MUCH

314
00:15:41,040 --> 00:15:41,707
APPRECIATED.

315
00:15:41,707 --> 00:15:46,179
YOUR HUMAN, YOUR KINDNESS, YOUR

316
00:15:46,179 --> 00:15:48,915
INSIGHTFUL SUPPORT HAVE ENRICHED

317
00:15:48,915 --> 00:15:52,652
OUR LIVES IN WAYS I CANNOT FULLY

318
00:15:52,652 --> 00:15:53,753
EXPRESS.

319
00:15:53,753 --> 00:15:58,024
NOW AS YOU STEP INTO THIS NEW

320
00:15:58,024 --> 00:16:03,930
CHAPTER OF LIFE, I'M EXCITED FOR

321
00:16:03,930 --> 00:16:04,463
YOU.

322
00:16:04,463 --> 00:16:05,665
ALAN, YOU WILL SEE, RETIREMENT

323
00:16:05,665 --> 00:16:06,666
IS NOT AN END.

324
00:16:06,666 --> 00:16:08,301
IT IS A BEGINNING.

325
00:16:08,301 --> 00:16:12,238
IT IS A TIME TO EXPLORE NEW

326
00:16:12,238 --> 00:16:15,975
PASSIONS, NEW HOBBIES, TO

327
00:16:15,975 --> 00:16:20,012
TRAVEL, TO SPEND TIME WITH

328
00:16:20,012 --> 00:16:21,547
FRIENDS, FAMILY, ENJOY NATURE

329
00:16:21,547 --> 00:16:24,717
INCLUDING BIRD WATCHING AND ALL

330
00:16:24,717 --> 00:16:25,518
THE BEAUTY.

331
00:16:25,518 --> 00:16:28,321
I KNOW WHATEVER YOU DECIDE TO

332
00:16:28,321 --> 00:16:33,025
PURSUE YOU WILL APPROACH IT WITH

333
00:16:33,025 --> 00:16:36,095
THE SAME DEDICATION THAT YOU

334
00:16:36,095 --> 00:16:39,065
BROUGHT TO YOUR SCIENTIFIC

335
00:16:39,065 --> 00:16:39,498
CAREER.

336
00:16:39,498 --> 00:16:43,102
ALAN, THANKS FOR BEING NOT ONLY

337
00:16:43,102 --> 00:16:46,372
A SCIENTIST AND MENTOR, ALSO A

338
00:16:46,372 --> 00:16:48,908
TRUE AND LOYAL FRIEND.

339
00:16:48,908 --> 00:16:50,209
YOUR LEGACY WILL LIVE IN THE

340
00:16:50,209 --> 00:16:52,511
HEART OF THOSE YOU HAVE

341
00:16:52,511 --> 00:16:55,681
INSPIRED, AND IN THE WORK THAT

342
00:16:55,681 --> 00:17:02,021
THEY WILL CONTINUE TO DO IN YOUR

343
00:17:02,021 --> 00:17:02,255
HONOR.

344
00:17:02,255 --> 00:17:04,757
WHILE YOU MAY BE RETIRING FROM

345
00:17:04,757 --> 00:17:05,524
PROFESSIONAL LIFE, YOU WILL

346
00:17:05,524 --> 00:17:08,394
ALWAYS BE PART OF OUR SCIENTIFIC

347
00:17:08,394 --> 00:17:10,663
COMMUNITY AND WE LOOK FORWARD TO

348
00:17:10,663 --> 00:17:13,799
SEEING WHERE THIS NEW JOURNEY

349
00:17:13,799 --> 00:17:14,533
TAKES YOU.

350
00:17:14,533 --> 00:17:18,471
MAY THE NEXT CHAPTER OF YOUR

351
00:17:18,471 --> 00:17:22,608
LIFE BE FILLED WITH JOY,

352
00:17:22,608 --> 00:17:25,378
ADVENTURE, AND ALL THE HAPPINESS

353
00:17:25,378 --> 00:17:28,648
YOU SO RICHLY DESERVE, ALAN.

354
00:17:28,648 --> 00:17:31,817
ENJOY, CHEERS TO YOU.

355
00:17:31,817 --> 00:17:42,028
THANK YOU. 

356
00:18:07,186 --> 00:18:14,226
>> THAT'S A HARD ACT TO
FOLLOW.

357
00:18:14,226 --> 00:18:15,428
I THANK YOU, FRANCOISE.

358
00:18:15,428 --> 00:18:18,197
AGAIN, I COULDN'T WISH FOR

359
00:18:18,197 --> 00:18:19,765
ANYTHING MORE.

360
00:18:19,765 --> 00:18:22,468
OUR NEXT SPEAKER IS JONATHAN

361
00:18:22,468 --> 00:18:28,975
STOYE, AND JUST A FEW WORDS.

362
00:18:28,975 --> 00:18:32,178
SO, AS FRANCOISE INDICATED, I

363
00:18:32,178 --> 00:18:34,580
CAME INTO MURINE LEUKEMIA VIRUS

364
00:18:34,580 --> 00:18:40,052
WORK BY WAY OF BOB BASSEN.

365
00:18:40,052 --> 00:18:42,121
I INHERITED BOB BASSEN AS A

366
00:18:42,121 --> 00:18:45,057
BOSS, WHEN I INHERITED BOB HE

367
00:18:45,057 --> 00:18:47,360
WAS ACTUALLY ON SABBATICAL IN

368
00:18:47,360 --> 00:18:47,793
PARIS.

369
00:18:47,793 --> 00:18:53,265
AND HE CAME BACK AND SAID, OKAY

370
00:18:53,265 --> 00:18:56,035
NOW, THERE'S THIS NEW THING

371
00:18:56,035 --> 00:18:57,503
Fv1 RESTRICTION, BRAND NEW AND

372
00:18:57,503 --> 00:18:58,337
REALLY INTERESTING.

373
00:18:58,337 --> 00:19:03,042
AND SO WE'RE GOING TO WORK ON

374
00:19:03,042 --> 00:19:03,242
IT.

375
00:19:03,242 --> 00:19:06,579
AND SO THIS -- WE NOW SEE Fv1

376
00:19:06,579 --> 00:19:11,717
AS A MEMBER OF THE CLASS OF

377
00:19:11,717 --> 00:19:13,319
RESTRICTION FACTORS, BUT AT THE

378
00:19:13,319 --> 00:19:17,590
TIME NOBODY HAD EVER HEARD OF A

379
00:19:17,590 --> 00:19:19,558
RESTRICTION FACTOR, Fv1 WAS

380
00:19:19,558 --> 00:19:23,062
UNIQUE, SINGULAR FOR MANY YEARS.

381
00:19:23,062 --> 00:19:26,899
I DID SOME -- IN BOB'S LAB I DID

382
00:19:26,899 --> 00:19:31,370
SOME KIND OF ELEMENTARY GENETICS

383
00:19:31,370 --> 00:19:33,572
CHARACTERIZING SOME OF THE

384
00:19:33,572 --> 00:19:37,076
PARTIES OF ITS INTERACTION WITH

385
00:19:37,076 --> 00:19:38,577
MURINE LEUKEMIA VIRUSES WHICH

386
00:19:38,577 --> 00:19:41,447
ARE FASCINATING, STILL NOT FULLY

387
00:19:41,447 --> 00:19:43,949
UNDERSTOOD. 

388
00:19:43,949 --> 00:19:45,384
BUT THEN IT GOT HARD.

389
00:19:45,384 --> 00:19:48,220
WHEN IT GOT HARD I WENT WITH

390
00:19:48,220 --> 00:19:50,189
OTHER THINGS TO DO.

391
00:19:50,189 --> 00:19:53,125
AND THAT'S WHERE JONATHAN COMES

392
00:19:53,125 --> 00:19:53,559
IN.

393
00:19:53,559 --> 00:19:55,661
JONATHAN DID NOT DROP Fv1 WHEN

394
00:19:55,661 --> 00:19:56,862
IT GOT HARD.

395
00:19:56,862 --> 00:19:59,832
HE STUCK WITH IT YEAR AFTER YEAR

396
00:19:59,832 --> 00:20:03,436
AFTER YEAR, AND BROKE IT OPEN.

397
00:20:03,436 --> 00:20:06,038
HE CLONED Fv1 AND HAS

398
00:20:06,038 --> 00:20:07,239
CONTINUED TO CHARACTERIZE Fv1

399
00:20:07,239 --> 00:20:09,775
AND NOW OTHER RESTRICT FACTORS

400
00:20:09,775 --> 00:20:10,743
AS WELL.

401
00:20:10,743 --> 00:20:14,346
AND HAS LEARNED A LOT OF AMAZING

402
00:20:14,346 --> 00:20:15,681
THINGS ABOUT Fv1.

403
00:20:15,681 --> 00:20:17,416
SO, JONATHAN, THANK YOU FOR

404
00:20:17,416 --> 00:20:17,650
COMING.

405
00:20:17,650 --> 00:20:28,194
I'M DELIGHTED YOU CAN BE HERE. 

406
00:20:33,099 --> 00:20:35,134
>> IT'S A GREAT APPRECIATE
TO BE

407
00:20:35,134 --> 00:20:35,601
IN FREDERICK.

408
00:20:35,601 --> 00:20:46,112
IT WAS A PLACE THAT I REMEMBER

409
00:20:49,615 --> 00:20:52,284
WITH PLEASURE.

410
00:20:52,284 --> 00:20:53,719
WE FIRST MET AT COLD SPRING

411
00:20:53,719 --> 00:20:55,454
HARBOR 45 YEARS AGO.

412
00:20:55,454 --> 00:21:00,059
CAN YOU FIND ALAN LOOKING SOBER 

413
00:21:00,059 --> 00:21:08,134
CAN YOU FIND ME PARTYING?

414
00:21:08,134 --> 00:21:08,400
[LAUGHTER]

415
00:21:08,400 --> 00:21:10,436
IS THERE ANYONE ELSE IN THE ROOM

416
00:21:10,436 --> 00:21:12,404
WHO CAN FIND THEMSELVES IN THESE

417
00:21:12,404 --> 00:21:12,738
PICTURES?

418
00:21:12,738 --> 00:21:15,274
IT'S INTERESTING. 

419
00:21:15,274 --> 00:21:17,443
SIX OF THE SPEAKERS TODAY WERE

420
00:21:17,443 --> 00:21:19,445
PRESENT AT THAT MEETING.

421
00:21:19,445 --> 00:21:29,955
AT LEAST SIX, BY MY COUNT SIX.

422
00:21:33,826 --> 00:21:38,931
I WANT TO TALK ABOUT Fv1.

423
00:21:38,931 --> 00:21:41,767
AND SHOW HOW STARTING WITH THE

424
00:21:41,767 --> 00:21:43,502
BIOLOGY THAT ALAN AND OTHERS

425
00:21:43,502 --> 00:21:45,704
WORKED ON IN THE '70s WE

426
00:21:45,704 --> 00:21:47,039
LEARNED A LOT ABOUT

427
00:21:47,039 --> 00:21:54,413
RETROVIRUSES, HOW THEY WORK.

428
00:21:54,413 --> 00:21:56,682
SO, PEOPLE THOUGHT RETRO CAUSES

429
00:21:56,682 --> 00:21:57,683
CAUSED CANCER, THERE WAS

430
00:21:57,683 --> 00:22:00,853
INTEREST IN GENES IN MICE THAT

431
00:22:00,853 --> 00:22:06,759
WOULD PROTECT AGAINST RETROVIRUS

432
00:22:06,759 --> 00:22:07,393
INFECTION.

433
00:22:07,393 --> 00:22:09,195
AND FORTUNATELY APPROXIMATELY

434
00:22:09,195 --> 00:22:10,162
INDEPENDENTLY THESE TWO

435
00:22:10,162 --> 00:22:11,897
GENTLEMEN, FRANK LILLY AND WALLY

436
00:22:11,897 --> 00:22:17,603
ROWE, DISCOVERED THE GENE CALLED

437
00:22:17,603 --> 00:22:18,037
Fv1.

438
00:22:18,037 --> 00:22:19,205
BASIC PROPERTIES ARE SUMMARIZED

439
00:22:19,205 --> 00:22:22,508
HERE, IN MICE THERE ARE TWO

440
00:22:22,508 --> 00:22:25,811
MAJOR ALLELES, A LITTLE N AND

441
00:22:25,811 --> 00:22:31,817
LITTLE B, THEY RESTRICT B AND N

442
00:22:31,817 --> 00:22:37,089
TROPIC VIRUSES RESPECTIVELY.

443
00:22:37,089 --> 00:22:38,724
THE SUSCEPTIBILITY DETERMINED IN

444
00:22:38,724 --> 00:22:41,927
MLV MAPS TO CAPSID, SPECIFICALLY

445
00:22:41,927 --> 00:22:45,397
AMINO ACID 110 IN THE CAPSID

446
00:22:45,397 --> 00:22:45,664
MOLECULE.

447
00:22:45,664 --> 00:22:48,901
AS WE'LL SEE, THERE ARE OTHER

448
00:22:48,901 --> 00:22:52,638
PARTS OF CAPSID THAT ARE

449
00:22:52,638 --> 00:22:58,978
INVOLVED, CAN INFLUENCE THIS

450
00:22:58,978 --> 00:22:59,278
INTERACTION.

451
00:22:59,278 --> 00:23:01,580
SO, WORK Fv1 IN THE '70s,

452
00:23:01,580 --> 00:23:03,015
THESE ARE SOME PAPERS THAT CAME

453
00:23:03,015 --> 00:23:07,386
FROM HIM AND FROM BOB BASSEN.

454
00:23:07,386 --> 00:23:10,256
I'LL DRAW YOUR ATTENTION TO THE

455
00:23:10,256 --> 00:23:13,659
BOTTOM RIGHT-HAND CORNER, WHICH

456
00:23:13,659 --> 00:23:17,129
FOR ME WAS -- BECAME KEY, THAT'S

457
00:23:17,129 --> 00:23:19,665
THAT YOU CAN OVERCOME Fv1

458
00:23:19,665 --> 00:23:21,400
RESTRICTION IF YOU INFECT WITH

459
00:23:21,400 --> 00:23:24,370
THE RIGHT VIRUS BUT THE CELL

460
00:23:24,370 --> 00:23:26,672
MAKING -- IF THE CELL IS MAKING

461
00:23:26,672 --> 00:23:28,841
THAT VIRUS IT STILL SHOWS Fv1

462
00:23:28,841 --> 00:23:32,344
RESTRICTION, I'LL RETURN TO THAT

463
00:23:32,344 --> 00:23:33,445
BRIEFLY LATER.

464
00:23:33,445 --> 00:23:35,748
SO, THIS SLIDE SUMMARIZES WHAT

465
00:23:35,748 --> 00:23:39,485
WE KNOW ABOUT OR WHERE Fv1

466
00:23:39,485 --> 00:23:39,818
WORKS.

467
00:23:39,818 --> 00:23:41,787
WORK FROM ALAN AND MANY OTHER

468
00:23:41,787 --> 00:23:45,291
PEOPLE, SOME IN THIS ROOM TODAY,

469
00:23:45,291 --> 00:23:47,259
DESCRIBING WHERE Fv1 ACTS.

470
00:23:47,259 --> 00:23:52,765
Fv1 SEEMS TO TARGET VIRUS AT A

471
00:23:52,765 --> 00:23:57,569
STAGE IN THE REPLICATION CYCLE

472
00:23:57,569 --> 00:23:59,204
UNCOATINGS OCCUR, SEEMS TO BLOCK

473
00:23:59,204 --> 00:24:00,739
MURINE IMPORT.

474
00:24:00,739 --> 00:24:02,942
IT DOES NOT INTERACT -- Fv1

475
00:24:02,942 --> 00:24:04,476
DOES NOT INTERACT WITH CAPSID ON

476
00:24:04,476 --> 00:24:14,653
THE WAY OUT.

477
00:24:14,653 --> 00:24:18,891
SO, WE WERE INTERESTED IN Fv1

478
00:24:18,891 --> 00:24:21,660
IN THE '70s.

479
00:24:21,660 --> 00:24:23,962
I WISH I COULD FIND THE

480
00:24:23,962 --> 00:24:24,863
ANNOUNCEMENT, THE PIECE OF PAPER

481
00:24:24,863 --> 00:24:27,666
ON WHICH IT WAS WRITTEN I WAS

482
00:24:27,666 --> 00:24:30,269
GOING TO TALK ABOUT A PAPER ON

483
00:24:30,269 --> 00:24:34,006
Fv1 BUT COULDN'T FIND IT.

484
00:24:34,006 --> 00:24:36,308
THE FACT THAT THERE SEEMS TO BE

485
00:24:36,308 --> 00:24:38,610
A MOUSE GENE, WORKS ON MOUSE

486
00:24:38,610 --> 00:24:40,913
VIRUSES, PEOPLE SORT OF LOST

487
00:24:40,913 --> 00:24:44,416
INTEREST IN THE SUBJECT.

488
00:24:44,416 --> 00:24:49,421
BUT I ALWAYS BELIEVED STRONGLY

489
00:24:49,421 --> 00:24:51,090
IN COMPARATIVE APPROACH,

490
00:24:51,090 --> 00:24:53,625
STUDYING HUMAN RETROVIRUSES BY

491
00:24:53,625 --> 00:24:55,561
STUDYING NON-HUMAN RETROVIRUSES

492
00:24:55,561 --> 00:24:56,362
IN NON-HUMAN SYSTEM.

493
00:24:56,362 --> 00:24:59,631
SO WHEN I HAD THE TUNED TO SET

494
00:24:59,631 --> 00:25:02,601
UP MY OWN LAB, I DECIDED TO TRY

495
00:25:02,601 --> 00:25:04,803
AND USE A POSITIONAL APPROACH,

496
00:25:04,803 --> 00:25:05,804
GENETIC APPROACH THAT RATHER

497
00:25:05,804 --> 00:25:06,538
APPEALED TO ME.

498
00:25:06,538 --> 00:25:08,941
WHEN I WAS A POSTDOC WITH JOHN

499
00:25:08,941 --> 00:25:11,010
COFFIN, TOGETHER WITH WAYNE

500
00:25:11,010 --> 00:25:13,212
FRANKEL, WE IDENTIFIED FOUR

501
00:25:13,212 --> 00:25:15,414
ENDOGENOUS RETROVIRUSES THAT

502
00:25:15,414 --> 00:25:17,916
WERE LINKED TO Fv1

503
00:25:17,916 --> 00:25:18,550
GENETICALLY.

504
00:25:18,550 --> 00:25:20,319
AND I DECIDED TO USE THOSE AS

505
00:25:20,319 --> 00:25:23,822
STARTING POINTS IN THE SEARCH

506
00:25:23,822 --> 00:25:26,125
FOR Fv1.

507
00:25:26,125 --> 00:25:28,293
FIRST GENERATING A BETTER

508
00:25:28,293 --> 00:25:30,796
GENETIC MAP, THEN MAKING YAC

509
00:25:30,796 --> 00:25:35,200
CLONES, LARGE CLONES FOR THE

510
00:25:35,200 --> 00:25:37,069
REGION AND INTRODUCE INTO CELLS

511
00:25:37,069 --> 00:25:39,571
AND SHOW WHICH ONES HAVE Fv1,

512
00:25:39,571 --> 00:25:45,911
AND FOLLOW THE ACTIVITY.

513
00:25:45,911 --> 00:25:46,879
IT WAS A STRUGGLE.

514
00:25:46,879 --> 00:25:51,150
THERE WERE A NUMBER OF

515
00:25:51,150 --> 00:25:53,118
DIFFICULTIES, LIKE A MASS

516
00:25:53,118 --> 00:25:54,987
HEPATITIS VIRUS INFECTION OF THE

517
00:25:54,987 --> 00:25:56,488
ANIMAL COLONY, WE HAD TO DO LESS

518
00:25:56,488 --> 00:26:00,559
OF A SURVEY OF THE GENETIC

519
00:26:00,559 --> 00:26:01,660
CROSS.

520
00:26:01,660 --> 00:26:02,961
AND THERE WERE INSTABILITY REACT

521
00:26:02,961 --> 00:26:04,496
CLONES, AND OTHER THINGS OF THE

522
00:26:04,496 --> 00:26:08,000
EVENTUALLY WE CLONED THE GENE.

523
00:26:08,000 --> 00:26:08,801
GREAT.

524
00:26:08,801 --> 00:26:10,102
THIS WAS MY MOMENT IN THE

525
00:26:10,102 --> 00:26:10,369
SPOTLIGHT.

526
00:26:10,369 --> 00:26:12,504
DID I HAVE A MOMENT IN THE

527
00:26:12,504 --> 00:26:13,939
SPOTLIGHT.

528
00:26:13,939 --> 00:26:16,008
I RATHER ENJOYED IT.

529
00:26:16,008 --> 00:26:16,742
[LAUGHTER]

530
00:26:16,742 --> 00:26:19,378
IT TURNED OUT THAT Fv1 WAS

531
00:26:19,378 --> 00:26:22,147
DERIVED FROM ONE OF THESE

532
00:26:22,147 --> 00:26:22,681
ENDOGENOUS RETROVIRUSES.

533
00:26:22,681 --> 00:26:25,050
I WAS TRYING TO GET AWAY FROM

534
00:26:25,050 --> 00:26:26,819
ENDOGENOUS RETROVIRUS, SPENT SO

535
00:26:26,819 --> 00:26:29,354
MUCH TIME WORKING ON THEM, AND

536
00:26:29,354 --> 00:26:31,323
NOW ANOTHER ENDOGENOUS

537
00:26:31,323 --> 00:26:31,623
RETROVIRUS.

538
00:26:31,623 --> 00:26:38,096
IT IS A VIRUS BELONGING TO THE

539
00:26:38,096 --> 00:26:39,498
ERV-L, RISEN BY COLLEAGUES,

540
00:26:39,498 --> 00:26:42,134
Fv1 CAN BE RECOGNIZED AS OPEN

541
00:26:42,134 --> 00:26:43,902
READING FRAME OF 1400

542
00:26:43,902 --> 00:26:47,739
NUCLEOTIDES THAT ENCODES THE

543
00:26:47,739 --> 00:26:50,676
ABILITY TO RESTRICT VIRUSES.

544
00:26:50,676 --> 00:26:53,879
WE STILL DON'T KNOW WHERE --

545
00:26:53,879 --> 00:26:56,048
WHAT THE ORIGINAL ERV-L WAS.

546
00:26:56,048 --> 00:27:04,056
WE CAN FIND OTHERS IN GENOMES OF

547
00:27:04,056 --> 00:27:06,125
RODENTS, BUT 50% SIMILARITY.

548
00:27:06,125 --> 00:27:08,760
IT'S ONLY ABOUT 10% SIMILARITY,

549
00:27:08,760 --> 00:27:11,697
IF ONE IS INVOKING SOME KIND OF

550
00:27:11,697 --> 00:27:14,766
LIKE WITH LIKE MECHANISM, IT'S

551
00:27:14,766 --> 00:27:17,269
GOT TO BE SEEING SOMETHING, A

552
00:27:17,269 --> 00:27:19,471
MORE COMPLICATED PRINCIPLE THAN

553
00:27:19,471 --> 00:27:21,306
THE AMINO ACID SEQUENCE.

554
00:27:21,306 --> 00:27:24,409
SO, WE ALSO NOTED IN THE

555
00:27:24,409 --> 00:27:25,477
ORIGINAL CLONING THAT Fv1 DID

556
00:27:25,477 --> 00:27:27,913
NOT SEEM TO BE PRESENT IN RAT

557
00:27:27,913 --> 00:27:30,516
DNA, AND SO WE CONCLUDED THAT

558
00:27:30,516 --> 00:27:34,119
THE GENE HAD -- THIS Fv1 HAD

559
00:27:34,119 --> 00:27:35,587
ARISEN LESS THAN 10 MILLION

560
00:27:35,587 --> 00:27:37,556
YEARS AGO, SEPARATION BETWEEN

561
00:27:37,556 --> 00:27:39,191
RAT AND MOUSE.

562
00:27:39,191 --> 00:27:42,561
HOWEVER, A MORE EXTENSIVE STUDY

563
00:27:42,561 --> 00:27:46,765
OF LOTS OF DIFFERENT -- DNA FROM

564
00:27:46,765 --> 00:27:50,469
LOTS OF CREATURES SUGGESTED THAT

565
00:27:50,469 --> 00:27:52,971
IN FACT THE Fv1 INSERTION THAT

566
00:27:52,971 --> 00:27:58,043
GAVE RISE TO Fv1 OCCURRED 45

567
00:27:58,043 --> 00:27:58,777
MILLION YEARS AGO.

568
00:27:58,777 --> 00:28:01,513
IT WAS LOST BY DELETION OF THE

569
00:28:01,513 --> 00:28:06,218
FAIRLY HIGH FREQUENCY OR BY

570
00:28:06,218 --> 00:28:07,319
POINT MUTATION.

571
00:28:07,319 --> 00:28:08,620
NEVERTHELESS IT'S QUITE

572
00:28:08,620 --> 00:28:10,155
REMARKABLE THIS OPEN READING

573
00:28:10,155 --> 00:28:11,590
FRAME HAS PERSISTED FOR THIS

574
00:28:11,590 --> 00:28:14,259
PERIOD OF TIME.

575
00:28:14,259 --> 00:28:16,628
SO, THE CALCULATION SAYS THAT IN

576
00:28:16,628 --> 00:28:19,131
45 MILLION YEARS AGO THERE'S AN

577
00:28:19,131 --> 00:28:21,133
ENORMOUSLY HIGH PROBABILITY THE

578
00:28:21,133 --> 00:28:23,502
GENE WILL BE LOST OVER TIME

579
00:28:23,502 --> 00:28:25,003
WHICH SUGGESTS IN ORDER TO

580
00:28:25,003 --> 00:28:26,471
MAINTAIN THE GENE YOU HAVE TO

581
00:28:26,471 --> 00:28:28,774
HAVE A SELECTIVE FORCE.

582
00:28:28,774 --> 00:28:32,377
THAT SAYS IT'S A GOOD THING.

583
00:28:32,377 --> 00:28:37,749
NOW, THIS RAISES A PROBLEM

584
00:28:37,749 --> 00:28:39,518
BECAUSE MLVs ONLY AROSE, LET'S

585
00:28:39,518 --> 00:28:40,819
SAY ABOUT 5 MILLION YEARS AGO.

586
00:28:40,819 --> 00:28:43,322
SO WHAT WAS HAPPENING IN THE

587
00:28:43,322 --> 00:28:44,990
PREVIOUS 40 MILLION YEARS?

588
00:28:44,990 --> 00:28:49,795
IS IT POSSIBLE THERE ARE OTHER

589
00:28:49,795 --> 00:28:51,763
VIRUSES AROUND -- THAT Fv1

590
00:28:51,763 --> 00:28:54,733
COULD  RESTRICT?

591
00:28:54,733 --> 00:28:57,035
AND THAT'S A QUESTION WE SPENT

592
00:28:57,035 --> 00:28:59,071
TIME WORKING ON, TRYING IDENTIFY

593
00:28:59,071 --> 00:29:02,808
OTHER VIRUSES THAT MIGHT BE

594
00:29:02,808 --> 00:29:05,010
RESTRICTED BY Fv1.

595
00:29:05,010 --> 00:29:07,946
AND WE'VE NOW TESTED THE

596
00:29:07,946 --> 00:29:10,315
RESTRICTION PROPERTIES OF 39

597
00:29:10,315 --> 00:29:14,553
Fv1 DERIVATIVES AGAINST 22

598
00:29:14,553 --> 00:29:14,786
VIRUSES.

599
00:29:14,786 --> 00:29:16,955
WITH RESTRICTION OF GAMMA

600
00:29:16,955 --> 00:29:20,359
RETROVIRUS IS MOST COMMON, WE'VE

601
00:29:20,359 --> 00:29:22,227
ALSO IDENTIFIED EXAMPLES OF

602
00:29:22,227 --> 00:29:23,996
ACTIVITY AGAINST MULTIPLE OTHER

603
00:29:23,996 --> 00:29:27,799
KINDS OF RETROVIRUSES INCLUDING

604
00:29:27,799 --> 00:29:29,668
ALPHAS, BETAS, DELTA, LENTI, AND

605
00:29:29,668 --> 00:29:30,435
FOAMY VIRUSES.

606
00:29:30,435 --> 00:29:32,604
I DON'T HAVE TIME TO GO INTO

607
00:29:32,604 --> 00:29:33,939
FOAMY VIRUSES BUT IT'S HAVING

608
00:29:33,939 --> 00:29:37,109
GIVEN WHAT WE KNOW ABOUT THE

609
00:29:37,109 --> 00:29:40,278
STRUCTURE OF FOAMY VIRUSES.

610
00:29:40,278 --> 00:29:42,948
THIS JUST SHOWS YOU A TREE OF

611
00:29:42,948 --> 00:29:46,551
Fv1 IN LOTS OF RODENTS, AND

612
00:29:46,551 --> 00:29:49,821
THE VIRUSES THAT CAN BE

613
00:29:49,821 --> 00:29:52,691
RESTRICTED BY THESE.

614
00:29:52,691 --> 00:29:56,728
SO, THERE ARE MULTIPLE EXAMPLES

615
00:29:56,728 --> 00:29:58,030
OF RESTRICTION.

616
00:29:58,030 --> 00:30:02,434
IT'S RARE TO SEE RESTRICTION OF

617
00:30:02,434 --> 00:30:07,205
ALL THE DIFFERENT VIRUSES BY A

618
00:30:07,205 --> 00:30:08,640
SINGLE Fv1 MOLECULE.

619
00:30:08,640 --> 00:30:11,143
BY COMBINING THEM WE CAN LOSE

620
00:30:11,143 --> 00:30:12,344
BOTH ACTIVITIES, TAKE THE AMINO

621
00:30:12,344 --> 00:30:13,578
ACID OF ONE, TRY TO MAKE

622
00:30:13,578 --> 00:30:15,113
SOMETHING THAT SEE LOTS OF

623
00:30:15,113 --> 00:30:17,315
VIRUSES, WE DON'T GET IT.

624
00:30:17,315 --> 00:30:18,950
WHICH SAYS THIS THING CAN

625
00:30:18,950 --> 00:30:20,585
RECOGNIZE LOTS OF DIFFERENT

626
00:30:20,585 --> 00:30:23,221
VIRUSES, BUT THERE ARE CHANGES

627
00:30:23,221 --> 00:30:33,565
THAT OCCUR, ASSOCIATED WITH THE

628
00:30:33,565 --> 00:30:35,133
DIFFERENT RESTRICTION

629
00:30:35,133 --> 00:30:35,467
SPECIFICITIES.

630
00:30:35,467 --> 00:30:38,437
AROSE 45 MILLION YEARS AGO, CAN

631
00:30:38,437 --> 00:30:40,972
SEE LOTS OF THINGS.

632
00:30:40,972 --> 00:30:44,910
SOMETIMES THESE SIGNAL AMINO

633
00:30:44,910 --> 00:30:46,545
ACID CHANGE IN Fv1 IS

634
00:30:46,545 --> 00:30:47,946
SUFFICIENT TO ALLOW RECOGNITION

635
00:30:47,946 --> 00:30:50,148
OF A DIFFERENT VIRUS.

636
00:30:50,148 --> 00:30:53,752
AND YOU HAVE TO REMEMBER THAT

637
00:30:53,752 --> 00:30:56,588
THIS IS A SINGLE AMINO ACID IN A

638
00:30:56,588 --> 00:30:58,657
PROTEIN THAT DIFFERS FROM ITS

639
00:30:58,657 --> 00:31:00,792
TARGET BY 50 TO 90%.

640
00:31:00,792 --> 00:31:03,395
SO, IT IS VERY -- Fv1 IS

641
00:31:03,395 --> 00:31:06,231
DIFFERENT FROM ITS TARGET WITH A

642
00:31:06,231 --> 00:31:07,766
SINGLE AMINO ACID MAKING A

643
00:31:07,766 --> 00:31:08,867
DIFFERENCE BETWEEN RESTRICTING

644
00:31:08,867 --> 00:31:09,768
AND NOT RESTRICTING.

645
00:31:09,768 --> 00:31:12,604
AND SOME BUT NOT ALL CASES WE

646
00:31:12,604 --> 00:31:13,371
CAN COMBINE MULTIPLE

647
00:31:13,371 --> 00:31:14,906
SPECIFICITIES, THERE'S AT LEAST

648
00:31:14,906 --> 00:31:22,981
ONE CASE WHERE YOU AFEAR HAVE

649
00:31:22,981 --> 00:31:24,416
DUPLICATED, YOU HAVE TWO, ONE

650
00:31:24,416 --> 00:31:27,486
CAN RESTRICT ONE VIRUS, ANOTHER

651
00:31:27,486 --> 00:31:29,454
CAN RESTRICT ANOTHER KIND OF

652
00:31:29,454 --> 00:31:32,524
VIRUS, SEEMS TO BE AN EXAMPLE OF

653
00:31:32,524 --> 00:31:33,925
THINGS Fv1 IS CAPABLE OFF.

654
00:31:33,925 --> 00:31:36,795
I WOULD ARGUE THIS GENE SURVIVED

655
00:31:36,795 --> 00:31:39,297
FOR ALL THIS TIME, IMPLIES A

656
00:31:39,297 --> 00:31:40,732
SERIES OF INFECTIONS BY

657
00:31:40,732 --> 00:31:42,033
DIFFERENT VIRUSES, ALL THROUGH

658
00:31:42,033 --> 00:31:43,568
OUR HISTORY.

659
00:31:43,568 --> 00:31:47,239
AND I GUESS THAT TO A

660
00:31:47,239 --> 00:31:48,173
RETROVIROLOGIST THAT'S NOT A

661
00:31:48,173 --> 00:31:50,709
SURPRISED BUT SOME PEOPLE THINK

662
00:31:50,709 --> 00:31:52,477
ABOUT VIRUSES APPEARING ONCE,

663
00:31:52,477 --> 00:31:53,779
AND CAUSING AIDS, AND, YOU KNOW,

664
00:31:53,779 --> 00:31:55,180
THEY DON'T THINK ABOUT THE PAST

665
00:31:55,180 --> 00:32:00,786
OR THE FUTURE IN THIS REGARD.

666
00:32:00,786 --> 00:32:03,021
SO HOW DOES Fv1 RESTRICT?

667
00:32:03,021 --> 00:32:06,458
UNFORTUNATELY WE STILL DON'T

668
00:32:06,458 --> 00:32:07,492
KNOW.

669
00:32:07,492 --> 00:32:10,061
HOWEVER MUCH WE WORK, WE DON'T

670
00:32:10,061 --> 00:32:12,364
KNOW HOW IT WORKS.

671
00:32:12,364 --> 00:32:17,502
WE CAN SHOW SPECIFIC RESPONDING

672
00:32:17,502 --> 00:32:19,471
OF Fv1 TO ASSEMBLE CAPSID.

673
00:32:19,471 --> 00:32:25,610
YOU CAN ASSEMBLE CAPSIDS ON THE

674
00:32:25,610 --> 00:32:27,813
TUBES IN THE LEFT-HAND CORNER,

675
00:32:27,813 --> 00:32:30,081
FORMS A HEXAGONAL ARRAY IN A WAY

676
00:32:30,081 --> 00:32:32,951
THAT'S SIMILAR TO THAT SEEN IN

677
00:32:32,951 --> 00:32:38,623
MATURE  VIROMES.

678
00:32:38,623 --> 00:32:45,063
WE CAN SHOW SPECIFICITY, SHOW

679
00:32:45,063 --> 00:32:49,467
SPECIFIC BINDING OF Fv1 TO

680
00:32:49,467 --> 00:32:51,670
B-MLV BUT NOT N-MLV.

681
00:32:51,670 --> 00:32:53,405
YOU DON'T GET BINDING THIS WAY

682
00:32:53,405 --> 00:32:55,407
IF YOU STICK THE CAPSID MOLECULE

683
00:32:55,407 --> 00:33:01,580
ON TO THESE AND TRY TO PULL DOWN

684
00:33:01,580 --> 00:33:04,216
THIS COMPLEXES WITH THESE TUBES.

685
00:33:04,216 --> 00:33:09,588
SO WE THINK IT'S RECOGNIZING

686
00:33:09,588 --> 00:33:11,656
SOMETHING VIRUS-LIKE.

687
00:33:11,656 --> 00:33:14,759
AND THIS SEEMS TO SHOW A CERTAIN

688
00:33:14,759 --> 00:33:16,695
SIMILARITY TO THAT OF THE

689
00:33:16,695 --> 00:33:18,897
MORE -- PERHAPS THE MORE FAMOUS

690
00:33:18,897 --> 00:33:20,432
RESTRICTION FACTOR WHERE PEOPLE

691
00:33:20,432 --> 00:33:23,935
HAVE BEEN ABLE TO SHOW THAT

692
00:33:23,935 --> 00:33:31,610
Fv1 BINDS TO THE CAUSE FOR

693
00:33:31,610 --> 00:33:33,245
VIRUSES BY MICROSCOPY.

694
00:33:33,245 --> 00:33:34,346
IT BINDS TO ASSEMBLIES.

695
00:33:34,346 --> 00:33:38,583
THAT WAS ONE PIECE OF

696
00:33:38,583 --> 00:33:39,484
INFORMATION.

697
00:33:39,484 --> 00:33:42,554
WE ALSO REALIZED FROM STUDIES OF

698
00:33:42,554 --> 00:33:47,792
DIFFERENT MLVs, SOME OF WHICH

699
00:33:47,792 --> 00:33:50,862
AROSE BY ESCAPE FROM Fv1, BUT

700
00:33:50,862 --> 00:33:52,597
THE BINDING DOMAINS ARE SPREAD

701
00:33:52,597 --> 00:33:57,435
ACROSS THE ENTIRE SURFACE OF THE

702
00:33:57,435 --> 00:34:00,472
MOLECULE, SO -- OR RATHER AMINO

703
00:34:00,472 --> 00:34:02,240
ACID DIFFERENCES THAT CAN

704
00:34:02,240 --> 00:34:03,108
INFLUENCE INTERACTION ARE SPREAD

705
00:34:03,108 --> 00:34:08,480
ALL OVER THE TOP OF THE

706
00:34:08,480 --> 00:34:08,914
MOLECULE.

707
00:34:08,914 --> 00:34:11,449
AGAIN, ARGUING AGAINST THE

708
00:34:11,449 --> 00:34:13,285
SINGLE POINT OF CONTACT BUT A

709
00:34:13,285 --> 00:34:15,820
WIDE POINT OF CONTACT WITH

710
00:34:15,820 --> 00:34:18,123
Fv1, ITS TARGET.

711
00:34:18,123 --> 00:34:21,426
WE'VE LOOKED IN SOME DETAIL AT

712
00:34:21,426 --> 00:34:24,596
THE SEQUENCES OF THESE DIFFERENT

713
00:34:24,596 --> 00:34:25,997
Fv1s WE'VE ISOLATED, THIS

714
00:34:25,997 --> 00:34:28,233
INITIAL -- THE INITIAL ANALYSIS

715
00:34:28,233 --> 00:34:31,036
WAS DONE WITH 19 SPECIES OF

716
00:34:31,036 --> 00:34:31,269
Fv1.

717
00:34:31,269 --> 00:34:32,904
WE'VE NOW LOOKED AT AT LEAST 39

718
00:34:32,904 --> 00:34:33,672
OR SO.

719
00:34:33,672 --> 00:34:36,942
WHAT WE SEE IS A SEQUENCE TENDS

720
00:34:36,942 --> 00:34:40,445
TO BE WELL CONSERVED, WITH

721
00:34:40,445 --> 00:34:44,950
DIFFERENCES CLUSTERING IN FOUR

722
00:34:44,950 --> 00:34:48,853
REGIONS, OF WHICH THREE, WHAT WE

723
00:34:48,853 --> 00:34:52,057
CALL VB AND VC AND VD,

724
00:34:52,057 --> 00:34:55,627
ASSOCIATED WITH THE DIFFERENCES

725
00:34:55,627 --> 00:34:59,164
BETWEEN THE N AND ALLELES OF

726
00:34:59,164 --> 00:34:59,531
Fv1.

727
00:34:59,531 --> 00:35:04,936
SO, YOU'VE GOT A CONSERVED

728
00:35:04,936 --> 00:35:06,037
SEQUENCE IMPLYING CONSERVED

729
00:35:06,037 --> 00:35:08,673
FUNCTION BUT CLUSTERS OF

730
00:35:08,673 --> 00:35:10,208
DIFFERENCES, POSSIBLY IMPLYING

731
00:35:10,208 --> 00:35:11,943
THESE ARE IMPORTANT FOR THE

732
00:35:11,943 --> 00:35:13,378
BINDING INTERACTION.

733
00:35:13,378 --> 00:35:16,781
AND THIS JUST SHOWS ONE  EXAMPLE

734
00:35:16,781 --> 00:35:18,984
OF CHANGES YOU CAN SEE IN ONE OF

735
00:35:18,984 --> 00:35:19,784
THESE REGIONS.

736
00:35:19,784 --> 00:35:21,586
WE HAVE ONE AMINO ACID CHANGE

737
00:35:21,586 --> 00:35:23,121
THAT'S IMPORTANT FOR THE

738
00:35:23,121 --> 00:35:26,625
DIFFERENCE BETWEEN Fv1 N AND

739
00:35:26,625 --> 00:35:28,360
B, A DIFFERENT ONE, SLIGHTLY

740
00:35:28,360 --> 00:35:31,229
DIFFERENT POSITION, WHICH IS

741
00:35:31,229 --> 00:35:35,100
IMPORTANT FOR THIRD SPECIFICITY

742
00:35:35,100 --> 00:35:36,468
OF Fv1.

743
00:35:36,468 --> 00:35:39,471
AND OTHER CHANGES THAT ARE

744
00:35:39,471 --> 00:35:46,478
IMPORTANT FOR RECOGNITION OF

745
00:35:46,478 --> 00:35:47,779
STILL MORE VIRUSES.

746
00:35:47,779 --> 00:35:49,080
THE DETAILS ARE PERHAPS NOT

747
00:35:49,080 --> 00:35:52,917
IMPORTANT BUT THE IDEA THAT YOU

748
00:35:52,917 --> 00:35:56,855
CAN HAVE CHANGES OF RECOGNITION

749
00:35:56,855 --> 00:36:00,158
PROPERTIES ASSOCIATED WITH

750
00:36:00,158 --> 00:36:05,730
SIGNAL AMINO ACIDS IS IMPORTANT.

751
00:36:05,730 --> 00:36:09,334
SO, ONE OF THE PROBLEMS IN OUR

752
00:36:09,334 --> 00:36:10,101
STUDIES Fv1 IS FLIPPING

753
00:36:10,101 --> 00:36:11,202
IMPOSSIBLE TO WORK WITH.

754
00:36:11,202 --> 00:36:15,140
IF YOU TRY EXPRESS IT IN

755
00:36:15,140 --> 00:36:17,008
BACTERIA, IT ALWAYS AGGREGATED.

756
00:36:17,008 --> 00:36:20,278
AND THIS MAY BE A REFLECTION,

757
00:36:20,278 --> 00:36:23,681
ITS INHERENT ABILITY TO

758
00:36:23,681 --> 00:36:25,984
AGGREGATE WITH VIRUSES, IT CAN

759
00:36:25,984 --> 00:36:29,387
ALSO AGGREGATE WITH ITSELF.

760
00:36:29,387 --> 00:36:32,557
SO, WE -- 25 YEARS WE'VE SPENT

761
00:36:32,557 --> 00:36:34,526
TRYING TO EXPRESS THIS THING SO

762
00:36:34,526 --> 00:36:39,664
WE COULD DO STRUCTURAL STUDIES.

763
00:36:39,664 --> 00:36:43,601
WHEN ALPHA CAME AROUND, WE

764
00:36:43,601 --> 00:36:45,770
RELUCTANTLY TRIED PUTTING OUR

765
00:36:45,770 --> 00:36:51,910
MOLECULE INTO ALPHAFOLD.

766
00:36:51,910 --> 00:36:53,678
THIS SHOWS THE STRUCTURE OF THE

767
00:36:53,678 --> 00:36:55,747
C-TERMINAL REGION OF Fv1.

768
00:36:55,747 --> 00:37:02,220
THE N-TERMINAL  REGION IS STATE

769
00:37:02,220 --> 00:37:02,654
 -- IS

770
00:37:02,654 --> 00:37:03,021
STRAIGHTFORWARD.

771
00:37:03,021 --> 00:37:04,522
THIS IS WELL SEPARATED FROM THE

772
00:37:04,522 --> 00:37:04,723
DIMER.

773
00:37:04,723 --> 00:37:07,492
WHAT YOU CAN SEE IF WE MAP IN

774
00:37:07,492 --> 00:37:09,794
THESE -- IF WE INDICATE THE

775
00:37:09,794 --> 00:37:15,066
POSITION OF THESE VARIABLE

776
00:37:15,066 --> 00:37:16,367
REGIONS, AVB, VC, AGAIN

777
00:37:16,367 --> 00:37:19,637
APPROACHING THE SURFACE OF ONE

778
00:37:19,637 --> 00:37:21,406
SIDE, IMPLYING TWO DINNER PLATES

779
00:37:21,406 --> 00:37:22,841
STICKING TO ONE ANOTHER.

780
00:37:22,841 --> 00:37:25,710
PERHAPS THIS IS IMPORTANT FOR

781
00:37:25,710 --> 00:37:26,010
RESTRICTION.

782
00:37:26,010 --> 00:37:31,049
WE TOOK THIS A BIT FURTHER VERY

783
00:37:31,049 --> 00:37:31,583
RECENTLY.

784
00:37:31,583 --> 00:37:33,251
I'M SORRY, THIS ISN'T A

785
00:37:33,251 --> 00:37:36,988
SUBMARINE OR SOMETHING BUT IT'S

786
00:37:36,988 --> 00:37:39,290
A MODEL OF THE CORE HEXAMER

787
00:37:39,290 --> 00:37:42,861
BASED ON WORK FROM ALAN AND

788
00:37:42,861 --> 00:37:46,030
COLLABORATORS, AND WE'VE TRIED

789
00:37:46,030 --> 00:37:47,165
TO ADD THIS Fv1 BINDING DOMAIN

790
00:37:47,165 --> 00:37:48,700
A TOP OF IT.

791
00:37:48,700 --> 00:37:50,768
WHAT WE CAN SEE WHEN WE DID

792
00:37:50,768 --> 00:37:52,203
THAT, THIS IS STILL VERY

793
00:37:52,203 --> 00:37:54,606
PRELIMINARY, AND WE WOULD LIKE

794
00:37:54,606 --> 00:37:58,109
TO HAVE MORE INFORMATION BY

795
00:37:58,109 --> 00:37:59,544
CROSS-LINKING OR MASS SPEC TYPE

796
00:37:59,544 --> 00:38:00,612
DATA APPROACHES.

797
00:38:00,612 --> 00:38:08,086
WHAT WE SEEM TO SEE IS E349 AND

798
00:38:08,086 --> 00:38:13,324
K358, TWO AMINO ACIDS I

799
00:38:13,324 --> 00:38:15,760
MENTIONED ARE BINDING TO OR

800
00:38:15,760 --> 00:38:19,664
ASSOCIATED WITH RESIDUES IN TWO

801
00:38:19,664 --> 00:38:24,602
DIFFERENT PARTS OF THE HEXAMER

802
00:38:24,602 --> 00:38:29,207
CONSIST END Fv1 IS SOMEHOW

803
00:38:29,207 --> 00:38:30,074
CROSS-LINKING DIFFERENT

804
00:38:30,074 --> 00:38:32,577
MOLECULES OF ITS TARGET.

805
00:38:32,577 --> 00:38:34,879
BUT IT IS VERY EARLY DAYS,

806
00:38:34,879 --> 00:38:40,585
MODELING IS STILL VERY

807
00:38:40,585 --> 00:38:42,320
PRELIMINARY, IT'S DIFFICULT, BUT

808
00:38:42,320 --> 00:38:46,925
WE WILL CONTINUE TO PURSUE THAT.

809
00:38:46,925 --> 00:38:51,763
SO, Fv1 IS DERIVED FROM A

810
00:38:51,763 --> 00:38:55,833
RETROVIRUS, DOESN'T APPEAR TO

811
00:38:55,833 --> 00:38:58,369
INTERACT WITH SINGLE CA

812
00:38:58,369 --> 00:39:00,205
MOLECULES, WE'VE DONE THAT IN

813
00:39:00,205 --> 00:39:00,772
CELLS.

814
00:39:00,772 --> 00:39:03,541
WE KNOW THAT Fv1 CAN FORM

815
00:39:03,541 --> 00:39:06,811
PUNCTATE ASSEMBLIES AND THAT IF

816
00:39:06,811 --> 00:39:08,880
WE PREVENT THAT WE CAN PREVENT

817
00:39:08,880 --> 00:39:09,180
RESTRICTION.

818
00:39:09,180 --> 00:39:14,586
SO THE IDEA THERE'S SOME HIGHER

819
00:39:14,586 --> 00:39:15,687
CAPSID-LIKE CAPSID INTERACTION

820
00:39:15,687 --> 00:39:17,755
TAKING PLACE SEEMS REASONABLE.

821
00:39:17,755 --> 00:39:20,358
AND WE WOULD POSTULATE IT ACTED

822
00:39:20,358 --> 00:39:22,026
BY PREVENTING CROSS-LINKING IN A

823
00:39:22,026 --> 00:39:25,630
WAY YOU CAN INHIBIT HIV BY

824
00:39:25,630 --> 00:39:28,366
PUTTING IN CYSTEINE RESIDUES IN

825
00:39:28,366 --> 00:39:32,170
THE RIGHT PLACES, CROSS-LINK THE

826
00:39:32,170 --> 00:39:32,403
HEXAMER.

827
00:39:32,403 --> 00:39:37,875
I'M GOING TO CHANGE GEARS AND

828
00:39:37,875 --> 00:39:40,478
BRIEFLY MENTION SOME OTHER

829
00:39:40,478 --> 00:39:43,781
FACTORS IN WHICH THE GAG GENE IS

830
00:39:43,781 --> 00:39:46,084
INVOLVED IN RESTRICTION.

831
00:39:46,084 --> 00:39:56,461
THE FIRST IS A FORMER

832
00:40:01,266 --> 00:40:02,734
RESTRICTION SEEN JAAGSIEKTE.

833
00:40:02,734 --> 00:40:07,205
IT APPEARS TO BE A DEFECTIVE

834
00:40:07,205 --> 00:40:08,706
GAGPOLY PROTEIN THAT INTERFERES

835
00:40:08,706 --> 00:40:10,608
WITH LATE REPLICATION OF THE

836
00:40:10,608 --> 00:40:11,409
RELATED VIRUSES.

837
00:40:11,409 --> 00:40:14,646
MAYBE ALAN CAN TELL US HOW THIS

838
00:40:14,646 --> 00:40:15,947
MIGHT WORK.

839
00:40:15,947 --> 00:40:19,250
AND SEEMS TO BE ASSOCIATED WITH

840
00:40:19,250 --> 00:40:20,585
TRAFFIC AT THE VIRUS GENE.

841
00:40:20,585 --> 00:40:23,755
THAT'S ONE CASE WHERE GAG IS

842
00:40:23,755 --> 00:40:29,460
IMPORTANT FOR RESTRICTION.

843
00:40:29,460 --> 00:40:35,566
ANOTHER IS IN THE YEAST, TY1,

844
00:40:35,566 --> 00:40:39,837
YEAST LIKE US DOESN'T WANT WILD

845
00:40:39,837 --> 00:40:40,471
VIRUS INTERFERING AND MUTATING.

846
00:40:40,471 --> 00:40:42,907
THEY HAVE A WAY OF REGULATING,

847
00:40:42,907 --> 00:40:46,110
SEEMS TO BE MEDIATED BY A

848
00:40:46,110 --> 00:40:47,178
PROTEIN THAT'S ENCODED WITHIN

849
00:40:47,178 --> 00:40:49,380
THE GAG GENE.

850
00:40:49,380 --> 00:40:52,550
AND IT INHIBITS VIRUS PARTICLE

851
00:40:52,550 --> 00:40:53,251
FORMATION.

852
00:40:53,251 --> 00:40:54,185
AND IN COLLABORATION WITH I

853
00:40:54,185 --> 00:40:57,121
THINK SOME OF YOUR FORMER

854
00:40:57,121 --> 00:40:57,722
COLLABORATORS, DAVID GARFINKEL

855
00:40:57,722 --> 00:41:00,992
WHO USED TO BE HERE, WE'VE DONE

856
00:41:00,992 --> 00:41:06,264
STRUCTURAL ANALYSES THAT WOULD

857
00:41:06,264 --> 00:41:09,734
SAY THAT THE INHIBITION ARISES

858
00:41:09,734 --> 00:41:11,269
BECAUSE YOU'RE APPROACHING AND

859
00:41:11,269 --> 00:41:12,804
COMPETING WITH ASSEMBLY PROCESS,

860
00:41:12,804 --> 00:41:17,008
SEEMS TO BE INVOLVING THE SAME

861
00:41:17,008 --> 00:41:20,144
RESIDUES AS WHEN AN MLV

862
00:41:20,144 --> 00:41:22,747
ASSEMBLES, THE SAME REGION OF

863
00:41:22,747 --> 00:41:27,952
C-TERMINAL REGION OF THE PROTEIN

864
00:41:27,952 --> 00:41:28,786
IMPORTANT FOR ASSEMBLY.

865
00:41:28,786 --> 00:41:31,656
I'M SURE THAT THERE'S BEEN MORE

866
00:41:31,656 --> 00:41:33,091
TALK ABOUT THESE GAG GENES

867
00:41:33,091 --> 00:41:40,398
ACTING AS SAFETY FACTORS.

868
00:41:40,398 --> 00:41:43,134
IT IS INTERESTING THAT GAG SEEMS

869
00:41:43,134 --> 00:41:45,670
TO BE INVOLVED IN MULTIPLE

870
00:41:45,670 --> 00:41:47,071
DIFFERENT WAYS, BECAUSE IT DOES

871
00:41:47,071 --> 00:41:54,078
DIFFERENT THINGS FOR THE VIRUS.

872
00:41:54,078 --> 00:42:00,651
THE CELL DOES USE  IT IN ORDER O

873
00:42:00,651 --> 00:42:01,586
INHIBIT VIRUS REPLICATION, ONE

874
00:42:01,586 --> 00:42:02,854
WONDERS HOW MANY MORE OF THESE

875
00:42:02,854 --> 00:42:05,456
THINGS ARE OUT THERE AND WHETHER

876
00:42:05,456 --> 00:42:06,457
OTHER GAGS PLAY PHYSIOLOGICAL

877
00:42:06,457 --> 00:42:08,059
ROLES OF ONE KIND OR ANOTHER,

878
00:42:08,059 --> 00:42:13,598
AND THIS REMAINS TO BE SEEN.

879
00:42:13,598 --> 00:42:16,100
I WANT TO CLOSE BY SAYING I TOO

880
00:42:16,100 --> 00:42:19,437
HAVE BECOME A RETIRED GENTLEMAN

881
00:42:19,437 --> 00:42:20,705
OF LEISURE.

882
00:42:20,705 --> 00:42:21,038
[LAUGHTER]

883
00:42:21,038 --> 00:42:25,209
MY LAB CLOSED, AND I BECAME AN

884
00:42:25,209 --> 00:42:27,278
EMERITUS SCIENTIST AT THE CRICK

885
00:42:27,278 --> 00:42:28,479
A FEW WEEKS AGO.

886
00:42:28,479 --> 00:42:32,450
THIS PICTURE WAS TAKEN ON TOP OF

887
00:42:32,450 --> 00:42:34,752
THE CRICK, JUST BEFORE COVID,

888
00:42:34,752 --> 00:42:37,955
I'VE LISTED HERE PEOPLE WHO DID

889
00:42:37,955 --> 00:42:38,923
TIME IN MY LAB.

890
00:42:38,923 --> 00:42:39,190
[LAUGHTER]

891
00:42:39,190 --> 00:42:40,792
I'D LIKE TO THANK THEM.

892
00:42:40,792 --> 00:42:44,729
I HOPE I HAVEN'T MISSED ANYBODY

893
00:42:44,729 --> 00:42:45,196
OUT.

894
00:42:45,196 --> 00:42:48,266
IT'S BEEN A WONDERFUL TIME.

895
00:42:48,266 --> 00:42:52,203
AND I'D LIKE TO CLOSE WITH A

896
00:42:52,203 --> 00:42:56,007
FINAL SLIDE OF A GATHERING OF

897
00:42:56,007 --> 00:42:56,407
RETROVIROLOGYISTS.

898
00:42:56,407 --> 00:42:57,675
ONE OF THE -- THIS HAS BEEN ONE

899
00:42:57,675 --> 00:43:00,945
OF THE NICE THINGS ABOUT BEING A

900
00:43:00,945 --> 00:43:02,480
MEMBER OF THIS GROUP, IT'S BEEN

901
00:43:02,480 --> 00:43:05,650
A PRIVILEGE TO BE PART OF THIS

902
00:43:05,650 --> 00:43:07,318
GROUP TRYING TO WORK OUT

903
00:43:07,318 --> 00:43:08,286
INTERESTING BIOLOGY.

904
00:43:08,286 --> 00:43:09,287
AND GETTING TOGETHER

905
00:43:09,287 --> 00:43:09,620
PERIODICALLY.

906
00:43:09,620 --> 00:43:12,223
SO WE'VE GOT STEVE, WE'VE GOT

907
00:43:12,223 --> 00:43:13,858
JOHN AND ALAN, MYSELF, IF YOU

908
00:43:13,858 --> 00:43:17,462
LOOK CAREFULLY YOU CAN FIND

909
00:43:17,462 --> 00:43:23,067
STEVE AND KARIN, I THINK MONICA.

910
00:43:23,067 --> 00:43:24,602
IT WOULD HAVE BEEN NICE TO HAVE

911
00:43:24,602 --> 00:43:26,604
ALL THE SPEAKERS IN ONE PLACE AT

912
00:43:26,604 --> 00:43:27,004
ONE TIME.

913
00:43:27,004 --> 00:43:28,739
I DON'T KNOW IF IT'S REASONABLE.

914
00:43:28,739 --> 00:43:30,508
ANYWAY, THANK YOU VERY MUCH.

915
00:43:30,508 --> 00:43:41,085
ALAN, IT'S BEEN A REAL PLEASURE 

916
00:43:51,963 --> 00:43:52,697
>> [OFF MICROPHONE].

917
00:43:52,697 --> 00:43:54,031
>> IT HAS, BUT IT'S NOT
VERY

918
00:43:54,031 --> 00:43:54,532
GOOD.

919
00:43:54,532 --> 00:43:55,900
IT'S NOT A VERY GOOD VIRUS.

920
00:43:55,900 --> 00:44:01,305
AND THE ONE THAT WE HAVE IS

921
00:44:01,305 --> 00:44:03,508
Fv1 IS AWAY FROM.

922
00:44:03,508 --> 00:44:07,078
WE TRIED TO DO THAT.

923
00:44:07,078 --> 00:44:11,782
THIS IS -- THEY HAVE

924
00:44:11,782 --> 00:44:13,351
RECONSTRUCTED THE VIRUS, THE

925
00:44:13,351 --> 00:44:15,653
TIME IS UP, PRETTY AWFUL,

926
00:44:15,653 --> 00:44:17,822
EXPERIMENTS ARE DIFFICULT.

927
00:44:17,822 --> 00:44:22,560
WE'VE TRIED. 

928
00:44:22,560 --> 00:44:23,928
>> YOU SPECULATE Fv1 MIGHT

929
00:44:23,928 --> 00:44:27,932
PREVENT ENCODING, IS IT POSSIBLE

930
00:44:27,932 --> 00:44:34,038
IT MIGHT, YOU KNOW, INDUCE

931
00:44:34,038 --> 00:44:34,739
PREMIUM ENCODING?

932
00:44:34,739 --> 00:44:39,510
>> SEQUESTERING IS A
REASONABLE

933
00:44:39,510 --> 00:44:39,810
POSSIBILITY.

934
00:44:39,810 --> 00:44:42,914
THERE ARE VERY OLD DATA FROM THE

935
00:44:42,914 --> 00:44:45,416
VIRUS LAB, SLIGHTLY DIFFICULT TO

936
00:44:45,416 --> 00:44:45,850
INTERPRET.

937
00:44:45,850 --> 00:44:48,486
WHICH SUGGESTS THAT FULL LENGTH

938
00:44:48,486 --> 00:44:50,788
DNA IS MADE NORMALLY AND GETS

939
00:44:50,788 --> 00:44:53,224
INTO THE NUCLEUS, BUT IT'S --

940
00:44:53,224 --> 00:44:58,863
THE DATA -- IT'S NOT A VERY

941
00:44:58,863 --> 00:45:04,835
CLEAR PAPER. 

942
00:45:04,835 --> 00:45:10,841
BUT YES, I MEAN, SO, WE KNOW

943
00:45:10,841 --> 00:45:15,880
TRIM5 IN SIMPLEST MANIFESTATION

944
00:45:15,880 --> 00:45:20,284
ACTS BY CAUSING PREMATURE

945
00:45:20,284 --> 00:45:22,687
DISASSEMBLY BUT IT'S CLEAR IF

946
00:45:22,687 --> 00:45:25,656
YOU BLOCK TRIM5 FROM ITS

947
00:45:25,656 --> 00:45:28,059
ACTIVITIES YOU KILL CAN BLOCK

948
00:45:28,059 --> 00:45:31,662
BOTH REPLICATION AND IT BECOMES

949
00:45:31,662 --> 00:45:42,006
MUCH CLOSER TO Fv1. 

950
00:45:44,141 --> 00:45:45,676
>> VERY NICE.

951
00:45:45,676 --> 00:45:49,647
FV EXPRESSED IN -- (INAUDIBLE).

952
00:45:49,647 --> 00:45:50,815
>> IT'S NOT INDUCIBLE BY

953
00:45:50,815 --> 00:45:51,148
INTERFERON.

954
00:45:51,148 --> 00:45:55,286
IT'S EXPRESSED ONLY IN VERY LOW

955
00:45:55,286 --> 00:45:55,553
LEVELS.

956
00:45:55,553 --> 00:45:58,155
AND I SUSPECT, I NEVER HAVE

957
00:45:58,155 --> 00:46:02,193
FORMALLY SHOWN THIS, IT MIGHT BE

958
00:46:02,193 --> 00:46:06,397
TOXIC IF OVEREXPRESSED.

959
00:46:06,397 --> 00:46:10,301
IT WAS INTERESTING WHEN WE FIRST

960
00:46:10,301 --> 00:46:11,836
CLONED Fv1, INTO PLASMID

961
00:46:11,836 --> 00:46:14,538
EXPRESSION VECTOR, THIS IS WORK

962
00:46:14,538 --> 00:46:23,848
DONE BY KATE BISHOP, SHE ASSAYED

963
00:46:23,848 --> 00:46:25,916
EIGHT CLONES WITH RESISTANCE.

964
00:46:25,916 --> 00:46:27,018
FIVE DELETED Fv1.

965
00:46:27,018 --> 00:46:29,420
SO WHICH IS WHY ALL THE ASSAYS

966
00:46:29,420 --> 00:46:32,490
WE DO NOW ARE BASED ON TRANSIENT

967
00:46:32,490 --> 00:46:32,923
EXPRESSION.

968
00:46:32,923 --> 00:46:35,559
AND IT'S CLEAR THAT

969
00:46:35,559 --> 00:46:36,761
OVEREXPRESSING CAN INTERFERE

970
00:46:36,761 --> 00:46:38,462
WITH A READOUT, COMPARED TO THE

971
00:46:38,462 --> 00:46:39,163
NATURAL LEVELS.

972
00:46:39,163 --> 00:46:44,435
IF YOU LOOK AT THE NATURAL

973
00:46:44,435 --> 00:46:52,410
LEVELS, YOU CAN BARELY SEE IT. 

974
00:46:52,410 --> 00:46:54,612
 

975
00:46:54,612 --> 00:46:57,148
 

976
00:46:57,148 --> 00:47:04,689
>> (INAUDIBLE) THIS STUDY

977
00:47:04,689 --> 00:47:11,796
SUGGESTS MOST OF THE RETROVIRUS

978
00:47:11,796 --> 00:47:15,633
IS ENDOGENOUS -- (INAUDIBLE).

979
00:47:15,633 --> 00:47:17,268
>> I DON'T KNOW.

980
00:47:17,268 --> 00:47:19,336
I MEAN, IF YOU WANT -- I DON'T

981
00:47:19,336 --> 00:47:21,972
THINK THERE WOULD BE ANY MARKET

982
00:47:21,972 --> 00:47:24,175
TRYING TO ENCOURAGE PEOPLE TO

983
00:47:24,175 --> 00:47:27,011
REINVESTIGATE PRECISELY WHERE

984
00:47:27,011 --> 00:47:28,479
THE TRANSCRIPTION OCCURS IN MLV

985
00:47:28,479 --> 00:47:30,715
BUT MAYBE IT WOULD BE

986
00:47:30,715 --> 00:47:31,015
INTERESTING.

987
00:47:31,015 --> 00:47:32,149
AGAIN, AN EXPERIMENT TO SEE.

988
00:47:32,149 --> 00:47:42,727
>> SO DOES IT ONLY WORK 
ON -- -

989
00:47:44,595 --> 00:47:45,296
(INAUDIBLE).

990
00:47:45,296 --> 00:47:51,535
>> THE FOAMY VIRUS IS THE

991
00:47:51,535 --> 00:47:53,270
INTERESTING ONE TO CONSIDER,

992
00:47:53,270 --> 00:47:55,673
BECAUSE THE DETERMINED

993
00:47:55,673 --> 00:47:59,276
SPECIFICITY FOR Fv1

994
00:47:59,276 --> 00:48:01,979
SPECIFICITY WITH FOAMY IN THE

995
00:48:01,979 --> 00:48:04,014
N-TERMINAL REGION, NOT IN THE

996
00:48:04,014 --> 00:48:04,381
CAPSID.

997
00:48:04,381 --> 00:48:07,184
WE KNOW THAT THIS FOAMY VIRUS IS

998
00:48:07,184 --> 00:48:08,152
NOT CLEAVED.

999
00:48:08,152 --> 00:48:15,259
AND WE KNOW THAT THIS THING --

1000
00:48:15,259 --> 00:48:21,198
OUTSIDE THE CAPSID, WHETHER IT'S

1001
00:48:21,198 --> 00:48:23,067
OBSCURED, INTERNAL REGIONS, OR

1002
00:48:23,067 --> 00:48:29,673
WHETHER THE N-TERMINAL REGION IS

1003
00:48:29,673 --> 00:48:30,941
ITSELF HELICAL, HEXAGONAL, I

1004
00:48:30,941 --> 00:48:31,509
DON'T KNOW.

1005
00:48:31,509 --> 00:48:33,778
IT WOULD BE VERY INTERESTING TO

1006
00:48:33,778 --> 00:48:34,345
LOOK AGAIN.

1007
00:48:34,345 --> 00:48:37,081
BUT PART OF THE PROBLEM IS THAT

1008
00:48:37,081 --> 00:48:39,049
WE CANNOT EXPRESS Fv1.

1009
00:48:39,049 --> 00:48:43,954
SO BINDING STUDIES ARE VERY

1010
00:48:43,954 --> 00:48:44,955
DIFFICULT.

1011
00:48:44,955 --> 00:48:47,057
NOW, WE HAVE -- EVERY FIVE YEARS

1012
00:48:47,057 --> 00:48:48,025
OR THREE YEARS, EVERY TIME WE

1013
00:48:48,025 --> 00:48:50,327
HAVE AN IDEA HOW WE MIGHT DO

1014
00:48:50,327 --> 00:48:54,064
THAT, WE'VE GOT ANOTHER ONE AT

1015
00:48:54,064 --> 00:49:00,404
THE MOMENT, BUT --

1016
00:49:00,404 --> 00:49:02,706
>> EXPRESSION MAY BE
DIFFICULT

1017
00:49:02,706 --> 00:49:08,045
BUT ALPHAFOLD DOES SEEM TO BE TO

1018
00:49:08,045 --> 00:49:11,448
DO INTERESTING THINGS.

1019
00:49:11,448 --> 00:49:13,617
MULTIMERS, HAVE YOU CONSIDERED

1020
00:49:13,617 --> 00:49:13,751
--

1021
00:49:13,751 --> 00:49:16,821
>> WE'VE CONSIDERED IT.

1022
00:49:16,821 --> 00:49:18,989
PART OF THE PROBLEM, WELL, NOT

1023
00:49:18,989 --> 00:49:20,324
WITH ALPHAFOLD ITSELF BUT WITH

1024
00:49:20,324 --> 00:49:24,461
THIS MOLECULE IS IT HAS THIS

1025
00:49:24,461 --> 00:49:30,434
LONG AND SPIRAL CODE AT THE

1026
00:49:30,434 --> 00:49:34,438
N-TERMINAL, THERE'S ABOUT 150

1027
00:49:34,438 --> 00:49:35,739
AMINO ACIDS OF UNSTRUCTURED

1028
00:49:35,739 --> 00:49:38,142
MATERIAL, AND THEN YOU HAVE THIS

1029
00:49:38,142 --> 00:49:42,079
DOMAIN THAT I SHOWED AT THE END

1030
00:49:42,079 --> 00:49:42,513
HERE.

1031
00:49:42,513 --> 00:49:48,452
NOW, IF -- AND I THINK  Fv1

1032
00:49:48,452 --> 00:49:54,024
MUST BE WORKING AT LEAST AS A

1033
00:49:54,024 --> 00:49:55,092
DIMER IF YOU DIMERIZE, THE WAY

1034
00:49:55,092 --> 00:49:57,294
YOU POSITION THE OTHER TWO, IT'S

1035
00:49:57,294 --> 00:49:57,661
DIFFICULT.

1036
00:49:57,661 --> 00:50:01,332
I SUSPECT -- NO, WE HAVE TO TRY,

1037
00:50:01,332 --> 00:50:02,099
TRY MORE.

1038
00:50:02,099 --> 00:50:05,703
MAYBE WE DO IN THEORY HAVE OUR

1039
00:50:05,703 --> 00:50:07,171
NEIGHBORS, PERHAPS I COULD

1040
00:50:07,171 --> 00:50:09,440
ENCOURAGE THEM TO TAKE ON THIS

1041
00:50:09,440 --> 00:50:13,944
AS A CHALLENGE, THAT THIS IS AN

1042
00:50:13,944 --> 00:50:15,913
INTERESTING MOLECULE.

1043
00:50:15,913 --> 00:50:18,115
BUT, NO, I AGREE.

1044
00:50:18,115 --> 00:50:19,650
WE HAVE ONLY JUST STARTED

1045
00:50:19,650 --> 00:50:22,253
THINKING ABOUT THIS.

1046
00:50:22,253 --> 00:50:25,222
WE HAVE THE ALPHAFOLD MODELS ARE

1047
00:50:25,222 --> 00:50:27,157
NOT -- ARE NOT NEW BUT TO TRY

1048
00:50:27,157 --> 00:50:32,229
AND FIT THIS AND PIN IT BY THE

1049
00:50:32,229 --> 00:50:36,467
AMINO ACIDS THAT WE HAVE FOR THE

1050
00:50:36,467 --> 00:50:38,168
VARIABLE REGIONS IS I THINK NEW,

1051
00:50:38,168 --> 00:50:40,804
TRYING TO DO THAT, WE JUST

1052
00:50:40,804 --> 00:50:46,243
STARTED. 

1053
00:50:46,243 --> 00:50:48,879
>> I'M MORE CONFUSED.

1054
00:50:48,879 --> 00:50:51,015
I WAS TOLD MLV DOESN'T GET INTO

1055
00:50:51,015 --> 00:50:52,883
THE NUCLEUS AND WAIT FOR THE

1056
00:50:52,883 --> 00:50:56,453
NUCLEAR ENVELOPE TO BREAK DOWN.

1057
00:50:56,453 --> 00:51:02,560
WHAT DOES IT MEAN, NUCLEAR

1058
00:51:02,560 --> 00:51:03,961
ENTRY?

1059
00:51:03,961 --> 00:51:05,729
>> WELL, IT BOOKS A LATE
STAGE

1060
00:51:05,729 --> 00:51:07,564
IN THAT PURPOSE WITH THE

1061
00:51:07,564 --> 00:51:10,768
REPLICATION CYCLE.

1062
00:51:10,768 --> 00:51:15,572
I'M SORRY. 

1063
00:51:15,572 --> 00:51:23,247
TEXT BOOKS FOR THE 1990s. 

1064
00:51:23,247 --> 00:51:25,849
I DON'T KNOW IF -- IF YOU

1065
00:51:25,849 --> 00:51:30,688
PREVENT THE CAPSID COMING APART.

1066
00:51:30,688 --> 00:51:33,857
>> RIGHT, YEAH.

1067
00:51:33,857 --> 00:51:35,926
>> WHICH IS COINCIDENT OR
JUST

1068
00:51:35,926 --> 00:51:38,462
PRECEDES NUCLEAR ENTRY, PERHAPS

1069
00:51:38,462 --> 00:51:41,065
YOU HAVE A SIMILAR PHENOTYPE TO

1070
00:51:41,065 --> 00:51:43,701
BLOCKING NUCLEAR ENTRY.

1071
00:51:43,701 --> 00:51:44,001
>> RIGHT.

1072
00:51:44,001 --> 00:51:45,869
YOU'RE LOOKING AT TWO CIRCLES.

1073
00:51:45,869 --> 00:51:47,972
I THINK THE ANSWER IS EVEN IN

1074
00:51:47,972 --> 00:51:49,707
RAPIDLY DIVIDING CELLS WHERE THE

1075
00:51:49,707 --> 00:51:53,377
NUCLEAR MEMBRANE IS BREAKING

1076
00:51:53,377 --> 00:51:54,611
DOWN, Fv1 BLOCKS WHAT WOULD

1077
00:51:54,611 --> 00:51:57,247
NORMALLY HAPPEN SO YOU DON'T GET

1078
00:51:57,247 --> 00:52:02,486
CIRCLES, YOU GET FULL LENGTH

1079
00:52:02,486 --> 00:52:02,720
LINEAR.

1080
00:52:02,720 --> 00:52:04,755
>> FULL LENGTH LINEAR, IN
THE

1081
00:52:04,755 --> 00:52:07,758
NUCLEUS, YOU KNOW, IN TERMS

1082
00:52:07,758 --> 00:52:18,302
OF -- IN THE NUCLEAR FRACTION. 

1083
00:52:23,741 --> 00:52:24,708
HELLO BACK, EVERYONE.

1084
00:52:24,708 --> 00:52:29,113
IT'S A PLEASURE NOW TO INTRODUCE

1085
00:52:29,113 --> 00:52:31,615
ONE OF THE WONDERFUL POSTDOCS

1086
00:52:31,615 --> 00:52:33,684
WHO HAS COME THROUGH MY LAB,

1087
00:52:33,684 --> 00:52:40,290
THAT'S DELPHINE MURIAUX. 

1088
00:52:40,290 --> 00:52:41,925
SHE CAME FROM FRANCE, AND SHE

1089
00:52:41,925 --> 00:52:49,033
WENT BACK TO FRANCE TWICE.

1090
00:52:49,033 --> 00:52:51,869
SHE HAD A PERMANENT JOB.

1091
00:52:51,869 --> 00:52:57,141
AND SHE WENT BACK TO TAKE THAT

1092
00:52:57,141 --> 00:52:59,343
PERMANENT JOB, BUT SHE CAME BACK

1093
00:52:59,343 --> 00:53:02,813
TO MY LAB, SUPPOSEDLY

1094
00:53:02,813 --> 00:53:04,915
TEMPORARILY, AND NOW SHE'S IN A

1095
00:53:04,915 --> 00:53:05,783
MUCH BETTER POSITION.

1096
00:53:05,783 --> 00:53:11,588
SHE NOW IS RUNNING A SHOP OF HER

1097
00:53:11,588 --> 00:53:13,457
OWN IN MONTPELLIER AND HAS DONE

1098
00:53:13,457 --> 00:53:18,295
A LOT OF WONDERFUL WORK ON HIV

1099
00:53:18,295 --> 00:53:19,696
ASSEMBLY, I THINK DOING A LOT OF

1100
00:53:19,696 --> 00:53:21,231
VIRAL PHYSICS AND I THINK WE'LL

1101
00:53:21,231 --> 00:53:24,101
HEAR A LITTLE ABOUT THAT TODAY.

1102
00:53:24,101 --> 00:53:25,736
DELPHINE, THANK YOU FOR COMING.

1103
00:53:25,736 --> 00:53:27,604
IT'S BEEN WONDERFUL KNOWING YOU

1104
00:53:27,604 --> 00:53:30,240
ALL THESE YEARS, AND LOOKING

1105
00:53:30,240 --> 00:53:40,617
FORWARD TO YOUR TALK. 

1106
00:53:41,385 --> 00:53:49,960
>> THANK YOU VERY MUCH,
ALAN.

1107
00:53:49,960 --> 00:53:52,763
HAVEN'T BEEN HERE FOR 20 YEARS,

1108
00:53:52,763 --> 00:53:56,200
AND THERE ARE MANY FRIENDS OF

1109
00:53:56,200 --> 00:53:59,369
MINE IN THIS ROOM, AND SO I

1110
00:53:59,369 --> 00:54:05,242
THANK A LOT THE ORGANIZER TO

1111
00:54:05,242 --> 00:54:06,477
INVITING ME TODAY.

1112
00:54:06,477 --> 00:54:08,879
I HOPE YOU ENJOY THE TALK.

1113
00:54:08,879 --> 00:54:15,252
ALAN, THANK YOU SO MUCH FOR ALL

1114
00:54:15,252 --> 00:54:19,056
THESE YEARS. 

1115
00:54:19,056 --> 00:54:22,025
SO I JUST PUT TO SHOW YOU MY

1116
00:54:22,025 --> 00:54:26,530
STAY IN ALAN'S LAB WAS ALWAYS

1117
00:54:26,530 --> 00:54:36,039
SECURE, I CAME BACK TO FRANCE

1118
00:54:36,039 --> 00:54:37,808
BUT DIDN'T LIKE IT.

1119
00:54:37,808 --> 00:54:47,985
TOO MUCH TEST, COULDN'T DO WHAT

1120
00:54:47,985 --> 00:54:51,788
I WANTED. 

1121
00:54:51,788 --> 00:54:57,528
I ASK IF I COULD COME BACK,

1122
00:54:57,528 --> 00:54:57,928
SURE.

1123
00:54:57,928 --> 00:55:00,464
I ROLLED BACK MY H1 VISA ON THE

1124
00:55:00,464 --> 00:55:00,697
SIDE.

1125
00:55:00,697 --> 00:55:02,533
I WEPT BACK TO FRANCE, TOOK THE

1126
00:55:02,533 --> 00:55:05,402
POSITION, I SAY KNOW, WENT BACK,

1127
00:55:05,402 --> 00:55:10,073
I TOOK BACK MY H1 VISA.

1128
00:55:10,073 --> 00:55:11,175
STAY FOR FOUR YEARS.

1129
00:55:11,175 --> 00:55:17,080
>> BE CAREFUL WHAT YOU
SAY.

1130
00:55:17,080 --> 00:55:20,050
>> AND THEN, YEAH, WHEN I
CAME

1131
00:55:20,050 --> 00:55:25,422
BACK TO FRANCE THE FIRST TIME I

1132
00:55:25,422 --> 00:55:26,390
HIDE IT.

1133
00:55:26,390 --> 00:55:27,591
THERE WAS THIS LITTLE GUY, I

1134
00:55:27,591 --> 00:55:30,227
DON'T KNOW WHO HE WAS, THAT SAY,

1135
00:55:30,227 --> 00:55:33,197
BUT I DON'T UNDERSTAND WHAT

1136
00:55:33,197 --> 00:55:35,832
HAPPENED IN 99.

1137
00:55:35,832 --> 00:55:38,035
I SAY, OKAY.

1138
00:55:38,035 --> 00:55:39,136
TWO CHOICE.

1139
00:55:39,136 --> 00:55:42,739
EITHER I LIE OR I PAY WITH

1140
00:55:42,739 --> 00:55:43,307
TRUTH.

1141
00:55:43,307 --> 00:55:45,142
IT WENT ONE, TWO, THREE.

1142
00:55:45,142 --> 00:55:46,810
WELL, IT'S TRUE, I WENT BACK TO

1143
00:55:46,810 --> 00:55:48,111
NIH BECAUSE I COULDN'T DO WHAT I

1144
00:55:48,111 --> 00:55:49,112
WANTED IN FRANCE.

1145
00:55:49,112 --> 00:55:50,214
BUT NOW I'M BACK.

1146
00:55:50,214 --> 00:55:52,916
IT'S UP TO YOU.

1147
00:55:52,916 --> 00:56:00,257
I GOT THE POSITION. 

1148
00:56:00,257 --> 00:56:03,560
FOR ME WAS RESTRAINING IN GO

1149
00:56:03,560 --> 00:56:08,699
BACK IN 1999, WE DID A LOT.

1150
00:56:08,699 --> 00:56:12,202
I TRIED TO SUMMARIZE.

1151
00:56:12,202 --> 00:56:16,573
IN 1995 IN COLD SPRING HARBOR,

1152
00:56:16,573 --> 00:56:19,109
IN 1996 MY FORMER BOSS INVITED

1153
00:56:19,109 --> 00:56:21,411
TO MY Ph.D. DEFENSE, YOU WERE

1154
00:56:21,411 --> 00:56:22,479
MY REVIEWER, RIGHT?

1155
00:56:22,479 --> 00:56:24,481
DURING MY Ph.D. DEFENSE.

1156
00:56:24,481 --> 00:56:29,820
AND AT THIS TIME WE SHARED THE

1157
00:56:29,820 --> 00:56:32,122
TEST FOR DIMERIZATION AND

1158
00:56:32,122 --> 00:56:33,457
NUCLEOCAPSID.

1159
00:56:33,457 --> 00:56:35,826
>> THAT'S WHAT YOU SAY.

1160
00:56:35,826 --> 00:56:36,093
[LAUGHTER]

1161
00:56:36,093 --> 00:56:38,795
I DIDN'T HEAR A WORD OF THE

1162
00:56:38,795 --> 00:56:39,930
DEFENSE BECAUSE IT WAS --

1163
00:56:39,930 --> 00:56:41,965
>> IT WAS IN FRENCH.

1164
00:56:41,965 --> 00:56:43,600
>> THE WINDOWS WERE OPEN,

1165
00:56:43,600 --> 00:56:44,835
BECAUSE IT WAS HOT.

1166
00:56:44,835 --> 00:56:47,571
>> IT WAS HOT.

1167
00:56:47,571 --> 00:56:49,740
>> EVERYBODY WAS TALKING
FRENCH

1168
00:56:49,740 --> 00:56:50,073
BESIDES.

1169
00:56:50,073 --> 00:56:50,741
>> YES.

1170
00:56:50,741 --> 00:56:53,810
BUT I DID ANSWER THE QUESTION IN

1171
00:56:53,810 --> 00:56:57,881
ENGLISH, NO? 

1172
00:56:57,881 --> 00:56:58,215
NOT PERFECTLY.

1173
00:56:58,215 --> 00:56:59,716
[LAUGHTER]

1174
00:56:59,716 --> 00:57:05,522
THE SLIDES WERE IN ENGLISH.

1175
00:57:05,522 --> 00:57:06,723
>> OKAY.

1176
00:57:06,723 --> 00:57:06,990
[LAUGHTER]

1177
00:57:06,990 --> 00:57:08,592
>> WE SHARE THE SAME
INTERESTS,

1178
00:57:08,592 --> 00:57:10,560
RIGHT?

1179
00:57:10,560 --> 00:57:11,995
THERE WAS A BIOCHEMIST, KNEW

1180
00:57:11,995 --> 00:57:13,830
NOTHING ABOUT VIROLOGY, I WAS

1181
00:57:13,830 --> 00:57:17,434
VERY HAPPY TO GET TO ALAN'S LAB.

1182
00:57:17,434 --> 00:57:19,436
HE REALLY TAUGHT ME VIROLOGY.

1183
00:57:19,436 --> 00:57:26,677
YOU'RE THE ONE WHO TAUGHT ME

1184
00:57:26,677 --> 00:57:27,778
VIROLOGY.

1185
00:57:27,778 --> 00:57:32,149
SO, I THINK WE TRIED ANOTHER

1186
00:57:32,149 --> 00:57:40,123
ROLE OF JUST GENOME PACKAGING.

1187
00:57:40,123 --> 00:57:44,828
AND THEN WE DID ALSO CONTINUED

1188
00:57:44,828 --> 00:57:47,464
TO WORK ON NUCLEOCAPSID MUTANT

1189
00:57:47,464 --> 00:57:53,937
AND FOUND THAT THERE WERE SOME

1190
00:57:53,937 --> 00:57:56,440
MUTANTS THAT WERE SEDATING RAG,

1191
00:57:56,440 --> 00:57:57,507
HIGHLIGHTING THE FACT

1192
00:57:57,507 --> 00:57:59,276
TRANSLATION AND PACKAGING WERE

1193
00:57:59,276 --> 00:58:00,811
LINKED TOGETHER.

1194
00:58:00,811 --> 00:58:05,182
AND WE TRIED TO UNDERSTAND THAT

1195
00:58:05,182 --> 00:58:07,284
BETTER. 

1196
00:58:07,284 --> 00:58:10,887
AND, YEAH, WE CONTINUED TO WORK

1197
00:58:10,887 --> 00:58:12,089
ON NUCLEOCAPSID ASSEMBLY AND

1198
00:58:12,089 --> 00:58:18,628
FOUND THAT IF YOU REMOVED THE

1199
00:58:18,628 --> 00:58:20,163
DOMAIN ACTIVITY PROTEIN WAS

1200
00:58:20,163 --> 00:58:22,132
STILL BINDING TO MEMBRANE BUT

1201
00:58:22,132 --> 00:58:27,738
WOULD NOT FORM STRUCTURES THAT

1202
00:58:27,738 --> 00:58:28,839
WERE LOOKING LIKE VIRAL

1203
00:58:28,839 --> 00:58:29,106
PARTICLE.

1204
00:58:29,106 --> 00:58:34,344
THAT WAS PART OF THE WORK WE

1205
00:58:34,344 --> 00:58:36,046
DID, AND OTHERS ALSO.

1206
00:58:36,046 --> 00:58:41,985
THEN I WANT TO SAY ALSO

1207
00:58:41,985 --> 00:58:44,054
OVERTHINKING, HAVE TIME TO GO AT

1208
00:58:44,054 --> 00:58:46,123
NIGHT WHEN EVERYBODY WAS GONE,

1209
00:58:46,123 --> 00:58:49,760
TALK TO THE BOSS, I WOULD GO

1210
00:58:49,760 --> 00:58:52,929
INTO HIS OFFICE.

1211
00:58:52,929 --> 00:58:56,767
THE NICE OFFICE.

1212
00:58:56,767 --> 00:58:58,668
[LAUGHTER] 

1213
00:58:58,668 --> 00:59:00,303
WE WERE ORGANIZED.

1214
00:59:00,303 --> 00:59:08,044
[LAUGHTER] 

1215
00:59:08,044 --> 00:59:10,147
AND THEN MY COLLEAGUE.

1216
00:59:10,147 --> 00:59:14,718
IT WAS REALLY AMAZING.

1217
00:59:14,718 --> 00:59:16,586
THAT'S ME PAINTING.

1218
00:59:16,586 --> 00:59:18,989
REALLY NICE.

1219
00:59:18,989 --> 00:59:22,926
SO I TRIED TO LIST MAYBE I

1220
00:59:22,926 --> 00:59:25,562
FORGET A FEW THINGS BUT ALAN

1221
00:59:25,562 --> 00:59:27,864
TAUGHT ME EVERYTHING ON MULV

1222
00:59:27,864 --> 00:59:31,234
INFECTION, TRIED TO SHOW ME TO

1223
00:59:31,234 --> 00:59:32,769
MAKE -- CONVINCE ME ABOUT

1224
00:59:32,769 --> 00:59:34,204
RESTRICTION FACTOR BUT I'M VERY

1225
00:59:34,204 --> 00:59:40,143
SORRY, I STILL DON'T UNDERSTAND.

1226
00:59:40,143 --> 00:59:40,677
[LAUGHTER]

1227
00:59:40,677 --> 00:59:43,447
AND YEAH, I'M SORRY, I'M SORRY.

1228
00:59:43,447 --> 00:59:53,990
YOU KNOW, I TRIED TO GET AWAY. 

1229
00:59:56,827 --> 01:00:03,667
IT WAS A CO-FACTOR. 

1230
01:00:03,667 --> 01:00:12,075
AND THEN I LEARNED TO DO VIRUS,

1231
01:00:12,075 --> 01:00:18,949
BUT YOU'RE THE ONE DOING THAT.

1232
01:00:18,949 --> 01:00:28,158
AND WE DID A LOT OF -- WHEN I GO

1233
01:00:28,158 --> 01:00:33,530
BACK TO FRANCE, THAT'S WHY I DID

1234
01:00:33,530 --> 01:00:33,797
ACTIVITY.

1235
01:00:33,797 --> 01:00:39,970
I WENT NOT TO DO THAT AGAIN.

1236
01:00:39,970 --> 01:00:50,480
AND ALSO I WANT TO THANK NCI,

1237
01:00:57,621 --> 01:01:00,423
TRAINING MANY METHODOLOGY.

1238
01:01:00,423 --> 01:01:00,757
WONDERFUL.

1239
01:01:00,757 --> 01:01:03,093
AND I THANK YOU FOR SCIENTIFIC

1240
01:01:03,093 --> 01:01:04,895
EDUCATION, LATE IN THE EVENING,

1241
01:01:04,895 --> 01:01:10,800
BECAUSE IN THE MORNING, WE'RE

1242
01:01:10,800 --> 01:01:12,235
NOT MORNING PERSONS.

1243
01:01:12,235 --> 01:01:14,504
AND TO HAVE BETTER ENGLISH, I

1244
01:01:14,504 --> 01:01:16,406
REMEMBER COMING WITH MY PAPER,

1245
01:01:16,406 --> 01:01:19,543
VERY HAPPY, AND THEN CAME BACK

1246
01:01:19,543 --> 01:01:23,547
ALL RED. 

1247
01:01:23,547 --> 01:01:25,482
SO BUT I TRIED.

1248
01:01:25,482 --> 01:01:27,551
BUT I'M NOT AS GOOD AS YOU.

1249
01:01:27,551 --> 01:01:28,885
YOU SHOULD WRITE A BOOK.

1250
01:01:28,885 --> 01:01:34,257
YOU SHOULD WRITE A BOOK.

1251
01:01:34,257 --> 01:01:35,992
AND ALSO MANY, MANY WE HAVE OUR

1252
01:01:35,992 --> 01:01:40,931
MEETING, THIS IS REALLY PART OF

1253
01:01:40,931 --> 01:01:46,303
MY FAMILY, I ENJOYED SEEING

1254
01:01:46,303 --> 01:01:48,939
TODAY. 

1255
01:01:48,939 --> 01:01:55,512
YOU TAUGHT ME THINKING, I THINK,

1256
01:01:55,512 --> 01:01:56,680
TRUST, SELF-EMPOWERMENT, DEAF

1257
01:01:56,680 --> 01:01:57,480
CONFIDENCE, TRUST.

1258
01:01:57,480 --> 01:02:01,084
THANK YOU FOR THAT.

1259
01:02:01,084 --> 01:02:03,053
AND ALSO WE HAD FRIENDSHIP,

1260
01:02:03,053 --> 01:02:06,423
RIGHT? 

1261
01:02:06,423 --> 01:02:09,192
AND WE WERE CELEBRATING ALL

1262
01:02:09,192 --> 01:02:11,695
TOGETHER, VERY FUN, ALAN IS NOT

1263
01:02:11,695 --> 01:02:16,600
ALWAYS SERIOUS, AS YOU CAN SEE

1264
01:02:16,600 --> 01:02:17,167
HERE.

1265
01:02:17,167 --> 01:02:18,802
[LAUGHTER]

1266
01:02:18,802 --> 01:02:22,405
AND STILL ON TODAY, WHO IS THIS

1267
01:02:22,405 --> 01:02:24,808
GUY BEHIND HERE?

1268
01:02:24,808 --> 01:02:26,142
THIS WAS THIS YEAR, COLD SPRING

1269
01:02:26,142 --> 01:02:29,412
HARBOR, VERY HAPPY AND WE HAD

1270
01:02:29,412 --> 01:02:33,483
THIS BIG PICTURE WITH ERIC,

1271
01:02:33,483 --> 01:02:37,053
FORMER TRAINEES, FROM THE

1272
01:02:37,053 --> 01:02:37,821
PROGRAMS, CONTINUING.

1273
01:02:37,821 --> 01:02:40,790
THANK YOU FOR ALL OF THAT.

1274
01:02:40,790 --> 01:02:46,496
AND THEN I TRAVELED BACK TO

1275
01:02:46,496 --> 01:02:47,130
FRANCE.

1276
01:02:47,130 --> 01:02:53,169
STILL CONTINUING THE WORK ON THE

1277
01:02:53,169 --> 01:02:55,472
ASSEMBLY, NUCLEOCAPSID, AND THEN

1278
01:02:55,472 --> 01:03:00,610
I TEN YEARS AGO, 12 YEARS AGO IN

1279
01:03:00,610 --> 01:03:02,245
MONTPELLIER TO GET CLOSER TO THE

1280
01:03:02,245 --> 01:03:04,648
VIRUS THAT I LIKE, THEY HAD A

1281
01:03:04,648 --> 01:03:10,387
VERY GOOD MICROSCOPE CENTER.

1282
01:03:10,387 --> 01:03:12,656
I END UP, BUT WHEN I WENT BACK

1283
01:03:12,656 --> 01:03:23,166
TO THE LAB I CONTINUED ON MULV

1284
01:03:25,335 --> 01:03:28,972
ASSEMBLY, SO WE DID THAT WORK,

1285
01:03:28,972 --> 01:03:37,981
WHERE WE SHOWED ALSO GAG WAS

1286
01:03:37,981 --> 01:03:40,183
TARGETING, THE DIFFERENCE WAS

1287
01:03:40,183 --> 01:03:45,188
THE FACT THAT THE FIRST SETTING

1288
01:03:45,188 --> 01:03:47,290
WAS SELECTING THE GAG AND

1289
01:03:47,290 --> 01:03:52,762
BINDING TO THE MEMBRANE BY P2,

1290
01:03:52,762 --> 01:03:55,932
WHICH DOESN'T NEED THAT, SO WE

1291
01:03:55,932 --> 01:03:58,635
CONTINUED THAT WORK.

1292
01:03:58,635 --> 01:04:02,405
AND GET INTERESTED IN THE LIPID

1293
01:04:02,405 --> 01:04:12,916
MEMBRANE, BINDING OF GAG, AND

1294
01:04:13,950 --> 01:04:14,584
RELATIONSHIP.

1295
01:04:14,584 --> 01:04:18,154
WITH ALAN ON THE INTERPLAY

1296
01:04:18,154 --> 01:04:25,061
BETWEEN GAG AND SERINE SITE,

1297
01:04:25,061 --> 01:04:34,137
ALSO A PROTEIN, THAT  -- SO THAK

1298
01:04:34,137 --> 01:04:34,404
YOU.

1299
01:04:34,404 --> 01:04:35,638
UNFORTUNATELY IN FRANCE COULDN'T

1300
01:04:35,638 --> 01:04:38,508
GET ANY FUNDING.

1301
01:04:38,508 --> 01:04:41,544
I HAD TO SWITCH TO HIV.

1302
01:04:41,544 --> 01:04:49,219
AND WHEN I MOVED TO MONTPELLIER,

1303
01:04:49,219 --> 01:04:52,622
WE END UP TOOLS AND DIFFERENT

1304
01:04:52,622 --> 01:04:54,157
SCALE OF INVESTIGATING INFECTED

1305
01:04:54,157 --> 01:04:59,162
CELLS, UP TO SINGLE VIRUSES BY

1306
01:04:59,162 --> 01:04:59,829
MICROSCOPY DEVICE, FLUORESCENT

1307
01:04:59,829 --> 01:05:02,899
DEVICE, AND WE END UP WORKING ON

1308
01:05:02,899 --> 01:05:07,270
THIS DIFFERENT VIRUSES OVER

1309
01:05:07,270 --> 01:05:07,604
TIME.

1310
01:05:07,604 --> 01:05:08,905
ESPECIALLY NUCLEAR RESCUE.

1311
01:05:08,905 --> 01:05:13,076
TODAY I WILL FOCUS ON HIV.

1312
01:05:13,076 --> 01:05:20,183
I END UP ALSO IN 2017 BEING HEAD

1313
01:05:20,183 --> 01:05:26,523
OF THE TEAM IN MONTPELLIER WITH

1314
01:05:26,523 --> 01:05:29,492
TRAINING AND IMAGING, I SPENT MY

1315
01:05:29,492 --> 01:05:31,361
TIME PUTTING ALL THIS

1316
01:05:31,361 --> 01:05:35,832
MICROSCOPY, NOW THERE'S NO MORE

1317
01:05:35,832 --> 01:05:37,267
ROOM ANYMORE.

1318
01:05:37,267 --> 01:05:38,668
TO STUDY MAINLY HIV REPLICATION

1319
01:05:38,668 --> 01:05:39,669
IN T CELLS.

1320
01:05:39,669 --> 01:05:42,005
SO IN THE LAB WE'RE MAINLY

1321
01:05:42,005 --> 01:05:50,079
INTERESTED IN T CELLS.

1322
01:05:50,079 --> 01:05:53,283
FLUORESCENT HIV.

1323
01:05:53,283 --> 01:05:55,051
TO STUDY THAT.

1324
01:05:55,051 --> 01:05:56,986
SO, WE ARE INTERESTED IN LATE

1325
01:05:56,986 --> 01:06:02,792
PHASE OF THE HIV REPLICATION, SO

1326
01:06:02,792 --> 01:06:05,862
AS I SAY, WORK VERY INSPIRING TO

1327
01:06:05,862 --> 01:06:06,362
ME.

1328
01:06:06,362 --> 01:06:08,932
YOU CAN SEE DIRECTLY I'M ON THE

1329
01:06:08,932 --> 01:06:12,468
LINE, AND WE'RE INTERESTED IN

1330
01:06:12,468 --> 01:06:14,604
WHAT P2 CLUSTERING BY GAG WOULD

1331
01:06:14,604 --> 01:06:21,544
TRIGGER IN A T CELL. 

1332
01:06:21,544 --> 01:06:26,883
WHAT THE ACTING UNDERNEATH

1333
01:06:26,883 --> 01:06:31,921
REGULATION CHANGE, AND HOW THIS

1334
01:06:31,921 --> 01:06:37,293
WOULD INTERFERE WITH THE NORMAL

1335
01:06:37,293 --> 01:06:41,464
BRAIN, THE FORMATION OR DOING

1336
01:06:41,464 --> 01:06:44,534
HIV PREDICTION.

1337
01:06:44,534 --> 01:06:48,404
SO MUCH INSPIRED BY THE WORK IN

1338
01:06:48,404 --> 01:06:49,172
ERIC FREED'S LAB, TOGETHER

1339
01:06:49,172 --> 01:06:52,642
POSTDOC AT THE SAME TIME, THIS

1340
01:06:52,642 --> 01:06:55,078
WAS THE PAPER THAT INSPIRE US.

1341
01:06:55,078 --> 01:06:57,680
BUT AT THAT PERIOD OF TIME,

1342
01:06:57,680 --> 01:06:59,983
CORRECT ME, ERIC, WE DIDN'T KNOW

1343
01:06:59,983 --> 01:07:03,686
REALLY GAG WAS BINDING TO

1344
01:07:03,686 --> 01:07:05,355
PREEXISTING P2 HUB DOMAINS IN

1345
01:07:05,355 --> 01:07:08,992
THE MEMBRANE, OR IF GAG WAS

1346
01:07:08,992 --> 01:07:12,295
GENERATING THAT.

1347
01:07:12,295 --> 01:07:18,201
SO, THAT'S WHAT -- YEAH,

1348
01:07:18,201 --> 01:07:24,674
INSPIRING ME A  LOT, LIPIDOMIC

1349
01:07:24,674 --> 01:07:26,542
EXTRACTION, COMPARING

1350
01:07:26,542 --> 01:07:27,510
COMPOSITION OF RETROVIRAL

1351
01:07:27,510 --> 01:07:29,379
PATHOGENS, AS GOING BACK TO

1352
01:07:29,379 --> 01:07:33,783
PLASMA MEMBRANE, INITIAL VERY

1353
01:07:33,783 --> 01:07:36,953
TYPICALLY THAT PSP 2 ENRICHED IN

1354
01:07:36,953 --> 01:07:39,155
THE MEMBRANE.

1355
01:07:39,155 --> 01:07:44,627
SO HOW COME? 

1356
01:07:44,627 --> 01:07:50,633
SO, WHAT WE DID, WE SET UP

1357
01:07:50,633 --> 01:07:54,570
TOOLS, BASICALLY SOME TOOLS, AND

1358
01:07:54,570 --> 01:08:00,143
WITH THE GAG PROTEIN, TRIED TO

1359
01:08:00,143 --> 01:08:02,779
MEASURE LIPID DOMAIN BY DYNAMICS

1360
01:08:02,779 --> 01:08:07,016
AND SEE IF GAG WAS A TRIGGER OF

1361
01:08:07,016 --> 01:08:11,421
THIS PHENOMENON. 

1362
01:08:11,421 --> 01:08:15,558
SO FOR EXAMPLE HERE FLAT

1363
01:08:15,558 --> 01:08:16,993
MEMBRANE, SCIENTIFIC COMPOSITION

1364
01:08:16,993 --> 01:08:19,529
UP TO PLASMA MEMBRANE, MANY

1365
01:08:19,529 --> 01:08:22,465
COMPOSITION, WE HAVE THE GAG

1366
01:08:22,465 --> 01:08:25,735
PROTEIN, THAT WE LABEL IN GREEN.

1367
01:08:25,735 --> 01:08:31,641
REPEAT P2 LABELED IN RED. 

1368
01:08:31,641 --> 01:08:36,579
YOU CAN SEE OVER TIME, YOU CAN

1369
01:08:36,579 --> 01:08:39,215
SEE THE P2 IS CLUSTERING OVER

1370
01:08:39,215 --> 01:08:39,449
TIME.

1371
01:08:39,449 --> 01:08:43,586
AND YOU HAVE PERFECT

1372
01:08:43,586 --> 01:08:46,422
CO-LOCALIZATION BETWEEN THIS

1373
01:08:46,422 --> 01:08:51,728
CLUSTER, SAYING THAT GAG WAS THE

1374
01:08:51,728 --> 01:08:54,831
TRIGGER OF THE CLUSTERING AND

1375
01:08:54,831 --> 01:08:58,034
SETTING MAYBE NOT AS FAR AS WE

1376
01:08:58,034 --> 01:09:00,236
CAN LOOK PREEXISTING DOMAIN.

1377
01:09:00,236 --> 01:09:02,105
SO GAG WAS GENERATING THIS

1378
01:09:02,105 --> 01:09:04,307
DOMAIN BUT WE HAD TO PROVE THAT

1379
01:09:04,307 --> 01:09:08,978
IN T CELLS, SO THAT'S HOW WE

1380
01:09:08,978 --> 01:09:11,714
ASSOCIATED WITH JACOB AND

1381
01:09:11,714 --> 01:09:18,654
CHRISTIAN, AND WE SHOWED THAT IF

1382
01:09:18,654 --> 01:09:20,690
WE TRANSFECT A T CELL WITH HIV

1383
01:09:20,690 --> 01:09:26,362
GAG PROTEIN ONLY OR IF WE INFECT

1384
01:09:26,362 --> 01:09:32,268
THE CELL WITH HIV CLONE WE CAN

1385
01:09:32,268 --> 01:09:36,339
SEE EACH DO THE AT RESOLUTION,

1386
01:09:36,339 --> 01:09:41,611
AND THEN WE LABELED THE T CELL

1387
01:09:41,611 --> 01:09:47,283
WITH DIFFERENT LIPID, LIKE P2,

1388
01:09:47,283 --> 01:09:50,353
FLUORESCENT, APPLIED COUPLING TO

1389
01:09:50,353 --> 01:09:51,921
MICROSCOPY AND MEASURE DYNAMIC

1390
01:09:51,921 --> 01:09:53,656
OF THE REPEAT OUTSIDE THE

1391
01:09:53,656 --> 01:09:57,026
BURDEN, INSIDE THE BURDEN.

1392
01:09:57,026 --> 01:10:00,763
FROM THAT WE CAN EXTRACT

1393
01:10:00,763 --> 01:10:02,198
CONFINEMENT INDEX OF LIPID.

1394
01:10:02,198 --> 01:10:03,933
WHAT YOU CAN SEE EVEN IN THE

1395
01:10:03,933 --> 01:10:08,204
NICE T CELL WE COULD SEE P2 WAS

1396
01:10:08,204 --> 01:10:12,909
HIGHLY TRAPPED BY THE GAG

1397
01:10:12,909 --> 01:10:15,445
PROTEIN, EITHER INFECTED CLONE

1398
01:10:15,445 --> 01:10:18,481
OR IN GAG EXPRESSION T CELL.

1399
01:10:18,481 --> 01:10:19,482
CHOLESTEROL ALSO.

1400
01:10:19,482 --> 01:10:20,483
CHOLESTEROL WERE MORE TRAPPED

1401
01:10:20,483 --> 01:10:26,823
WHEN YOU HAVE ENVELOPE.

1402
01:10:26,823 --> 01:10:32,395
DIDN'T FIND PE TRAPPING FOR P2

1403
01:10:32,395 --> 01:10:35,598
AND CHOLESTEROL, WHICH WAS

1404
01:10:35,598 --> 01:10:36,132
MATCHING.

1405
01:10:36,132 --> 01:10:40,403
WE SAY THAT P2 AND CHOLESTEROL

1406
01:10:40,403 --> 01:10:42,371
ACTUALLY ARE THE LIPID TRAPPED

1407
01:10:42,371 --> 01:10:44,774
BY INFECTED T CELL.

1408
01:10:44,774 --> 01:10:47,043
BUT THAT DOESN'T MEAN -- THE

1409
01:10:47,043 --> 01:10:51,013
PROTEIN BY ITSELF IS CONSTANTLY

1410
01:10:51,013 --> 01:10:54,283
CHANGING GRADIENT OF THE

1411
01:10:54,283 --> 01:10:55,718
MEMBRANE, THIS HAS STRONG

1412
01:10:55,718 --> 01:10:58,554
REPERCUSSION, P2 IS A MAIN

1413
01:10:58,554 --> 01:11:06,429
SIGNALING LIPID IN T CELL. 

1414
01:11:06,429 --> 01:11:10,600
IT CAN INDUCE DIVERSITY.

1415
01:11:10,600 --> 01:11:14,437
NOW I SPEND MY TIME TO TRY TO

1416
01:11:14,437 --> 01:11:18,508
IDENTIFY OR UNDERSTANDING WHICH

1417
01:11:18,508 --> 01:11:26,048
FACTOR ARE INVOLVED IN THIS

1418
01:11:26,048 --> 01:11:27,049
FORMATION.

1419
01:11:27,049 --> 01:11:29,418
AND HERE, BEFORE WE WERE PLUG

1420
01:11:29,418 --> 01:11:35,024
DOWN DIFFERENT PATHWAY OF

1421
01:11:35,024 --> 01:11:39,729
SIGNALING LIKE THE ONE, AND WE

1422
01:11:39,729 --> 01:11:41,697
SHOW THIS PATHWAY WAS REQUIRED

1423
01:11:41,697 --> 01:11:50,206
IN T CELL. 

1424
01:11:50,206 --> 01:11:54,944
YOU HAVE ONE, IRSP53, WE SHOW

1425
01:11:54,944 --> 01:11:58,548
THAT PAPER ACTIVATING WHEN GAG

1426
01:11:58,548 --> 01:11:59,982
WAS EXPRESSED.

1427
01:11:59,982 --> 01:12:06,656
SO THIS IS PARTICULAR, PROTEIN

1428
01:12:06,656 --> 01:12:13,529
IS ABLE TO GENERATE OUTWARD, AND

1429
01:12:13,529 --> 01:12:16,165
THE SIZE OF THIS PROTRUSION,

1430
01:12:16,165 --> 01:12:17,133
DIAMETER EQUIVALENT TO DIAMETER

1431
01:12:17,133 --> 01:12:19,669
OF A VIRUS.

1432
01:12:19,669 --> 01:12:30,213
SO, WE DECIDED TO STUDY IRSP 53

1433
01:12:31,447 --> 01:12:39,488
AND IRTKS, IN THE HIV CONTEXT.

1434
01:12:39,488 --> 01:12:43,626
THERE'S A POINTER.

1435
01:12:43,626 --> 01:12:44,727
OKAY.

1436
01:12:44,727 --> 01:12:49,532
YOU WILL BE ABLE TO SEE VIRUS

1437
01:12:49,532 --> 01:12:56,539
WAS REDUCED, AND HAD NOTHING, A

1438
01:12:56,539 --> 01:12:59,275
CLOSE PROTEIN AND COULD FIND BY

1439
01:12:59,275 --> 01:13:01,544
ELECTRON MICROSCOPY THANKS TO

1440
01:13:01,544 --> 01:13:05,815
PHILIP THAT THE BED WAS NOT

1441
01:13:05,815 --> 01:13:10,553
COMPLETE. 

1442
01:13:10,553 --> 01:13:12,855
IT WAS LIKE THIS, THE CELL WAS

1443
01:13:12,855 --> 01:13:17,560
NOT DYING, SO WE THOUGHT THAT

1444
01:13:17,560 --> 01:13:20,496
IRSP 53 HAD A ROLE IN ASSEMBLY,

1445
01:13:20,496 --> 01:13:22,898
TRIED TO SEE IF IT WAS WITH GAG,

1446
01:13:22,898 --> 01:13:25,534
AS YOU CAN SEE HERE IT WAS.

1447
01:13:25,534 --> 01:13:27,403
THEN WE DID DIFFERENT MUTANT OF

1448
01:13:27,403 --> 01:13:29,238
GAG AND TRIED TO IDENTIFY THE

1449
01:13:29,238 --> 01:13:32,008
DOMAIN REQUIRED FOR THIS

1450
01:13:32,008 --> 01:13:32,308
INTERACTION.

1451
01:13:32,308 --> 01:13:34,844
ACTUALLY I HAVE TO SAY SIGNAL

1452
01:13:34,844 --> 01:13:37,446
CLEAR, BUT WHAT WE CAN SEE IS

1453
01:13:37,446 --> 01:13:42,818
THAT IF WE HAVE THE HVR

1454
01:13:42,818 --> 01:13:48,424
RELATION, NOT BINDING TO THE

1455
01:13:48,424 --> 01:13:50,593
MEMBRANE, AND THE MUTATION, THE

1456
01:13:50,593 --> 01:13:50,927
INTERACTION.

1457
01:13:50,927 --> 01:13:55,331
IN OUR OPINION THIS INTERACTION

1458
01:13:55,331 --> 01:14:01,237
BETWEEN GAG AND IRSP 53 OCCUR AT

1459
01:14:01,237 --> 01:14:04,874
THE CELL MEMBRANE, AND WE REDUCE

1460
01:14:04,874 --> 01:14:12,214
A LITTLE BIT THIS INTERACTION SO

1461
01:14:12,214 --> 01:14:16,686
EITHER WAY, EITHER WE PULL THE

1462
01:14:16,686 --> 01:14:18,120
HYPOTHESIS IT OCCURS BEFORE BUT

1463
01:14:18,120 --> 01:14:19,422
HAVE NOT PROVEN THAT SO WE

1464
01:14:19,422 --> 01:14:24,126
CONTINUE TO WORK ON THAT.

1465
01:14:24,126 --> 01:14:29,632
THEN IF IRSP 53 WAS REQUIRED TO

1466
01:14:29,632 --> 01:14:33,903
THE SITE, WE USED MICROSCOPY, SO

1467
01:14:33,903 --> 01:14:39,275
THEN WE EXPRESSED THE GAG

1468
01:14:39,275 --> 01:14:41,243
PROTEIN THAT IS ABLE TO BRING,

1469
01:14:41,243 --> 01:14:44,080
AND YOU HAVE ONLY THE PLASMA

1470
01:14:44,080 --> 01:14:45,848
MEMBRANE UNDERNEATH THAT YOU CAN

1471
01:14:45,848 --> 01:14:52,722
SEE HERE.

1472
01:14:52,722 --> 01:14:53,956
FROM BLINKING, RESOLUTION

1473
01:14:53,956 --> 01:14:56,559
20-NANOMETER, PLATFORM IN GREEN.

1474
01:14:56,559 --> 01:15:03,032
WE CAN COME WITH THE ANTIBODY

1475
01:15:03,032 --> 01:15:05,768
LABELING IRSP 53 AND COMPARE

1476
01:15:05,768 --> 01:15:08,371
SIGNALING DIFFERENCE TO THE

1477
01:15:08,371 --> 01:15:10,940
BUDDING SITE.

1478
01:15:10,940 --> 01:15:14,210
THANKS TO CYRIL, AND HIS

1479
01:15:14,210 --> 01:15:15,411
WORKFLOW.

1480
01:15:15,411 --> 01:15:24,420
SO WE COULD DATA MINE THE CENTER

1481
01:15:24,420 --> 01:15:30,993
AND DISTANCE OF 300-NANOMETER,

1482
01:15:30,993 --> 01:15:35,931
FOR IRSP 53, INDEED IT IS

1483
01:15:35,931 --> 01:15:41,837
CLOSELY RELATED TO GAG ASSEMBLY

1484
01:15:41,837 --> 01:15:47,076
SIDE, SO YOU CAN SEE HERE, AND

1485
01:15:47,076 --> 01:15:55,418
ACTUALLY IT SEEMS TO SURROUNDING

1486
01:15:55,418 --> 01:15:58,154
THE ASSEMBLY BUDDING SITE, HAVE

1487
01:15:58,154 --> 01:15:59,455
THIS PATTERN.

1488
01:15:59,455 --> 01:16:03,058
SOME DON'T HAVE IT.

1489
01:16:03,058 --> 01:16:05,594
IT'S COMPLICATED SO WE BELIEVE

1490
01:16:05,594 --> 01:16:10,266
THAT THE TOP, THERE'S A TOP

1491
01:16:10,266 --> 01:16:11,767
RECRUITMENT OF IRSP 53 TO THE

1492
01:16:11,767 --> 01:16:22,311
BUDDING SITE AND DON'T KNOW THE

1493
01:16:23,279 --> 01:16:25,181
CORRELATION WITH THE PROTEIN.

1494
01:16:25,181 --> 01:16:29,985
WANTED IT FLOW IF IRSP 53 HAS

1495
01:16:29,985 --> 01:16:33,255
ANY ROLE IN GAG TO MEMBRANE

1496
01:16:33,255 --> 01:16:33,489
BINDING.

1497
01:16:33,489 --> 01:16:44,033
WE SET UP IN VITRO MODEL, WE HAD

1498
01:16:44,266 --> 01:16:48,237
LABEL WITH FLUEROSOL, GOOD

1499
01:16:48,237 --> 01:16:54,944
CODING IN P2, IRSP 53-I-BAR

1500
01:16:54,944 --> 01:16:59,782
DOMAIN, TO SEE MEMBRANE BINDING.

1501
01:16:59,782 --> 01:17:01,650
SO HERE OUR RESULT.

1502
01:17:01,650 --> 01:17:08,357
WE SHOW WHEN WE HAVE GAG, EVEN

1503
01:17:08,357 --> 01:17:13,062
IF CONTAIN P2, THERE'S NO MATCH

1504
01:17:13,062 --> 01:17:14,797
GAG BINDING.

1505
01:17:14,797 --> 01:17:25,241
IF YOU ADD THE I-BAR DOMAIN, WE

1506
01:17:25,241 --> 01:17:30,946
BELIEVE THE TWO PROTEINS WITH

1507
01:17:30,946 --> 01:17:31,380
INTERCONNECTED.

1508
01:17:31,380 --> 01:17:36,085
AND WE DID A MOVIE ON THAT.

1509
01:17:36,085 --> 01:17:44,460
SO HERE WE CAN SEE THE MEMBRANE.

1510
01:17:44,460 --> 01:17:46,595
IRSP53 I-BAR, AND IF YOU PUT BAG

1511
01:17:46,595 --> 01:17:49,532
ON IT, YOU CAN SEE THAT GAG

1512
01:17:49,532 --> 01:17:55,004
PREFERS TO BIND WHERE YOU HAVE

1513
01:17:55,004 --> 01:18:00,709
THE IRSP53 IN PINK. 

1514
01:18:00,709 --> 01:18:03,879
GAG PROTEIN LOCATE EXACTLY AT

1515
01:18:03,879 --> 01:18:05,848
THE TIP.

1516
01:18:05,848 --> 01:18:09,218
SO TWO PROTEIN ARE CERTAINLY

1517
01:18:09,218 --> 01:18:12,855
LINKED TOGETHER, AND WE TRIED TO

1518
01:18:12,855 --> 01:18:19,428
CONTINUE TO UNDERSTAND THIS

1519
01:18:19,428 --> 01:18:21,063
MOLECULAR PATHWAY.

1520
01:18:21,063 --> 01:18:25,434
THEN WE WANTED TO KNOW IF IT

1521
01:18:25,434 --> 01:18:28,504
COULD BE INCORPORATED IN GAG,

1522
01:18:28,504 --> 01:18:36,712
AND WE DID THAT WITH IRSP 53,

1523
01:18:36,712 --> 01:18:41,784
GFP, OR DOMAIN, OR PH-GFP.

1524
01:18:41,784 --> 01:18:45,588
YOU CAN SEE 53 IS THE MOST

1525
01:18:45,588 --> 01:18:49,491
INCORPORATED PROTEIN, AND COULD

1526
01:18:49,491 --> 01:18:52,394
FIND SOME IN THE HIV VIRUS BUT

1527
01:18:52,394 --> 01:18:56,899
VERY FEW IN HIV VIRUS SO MAYBE

1528
01:18:56,899 --> 01:18:58,200
THERE'S ANOTHER REGULATION HERE

1529
01:18:58,200 --> 01:19:05,541
THAT ALLOWS ITS JOB AT ONE POINT

1530
01:19:05,541 --> 01:19:11,413
BUT THERE IS A REGULATION WE

1531
01:19:11,413 --> 01:19:12,648
DON'T FULLY UNDERSTAND TODAY.

1532
01:19:12,648 --> 01:19:19,622
WE FINISH WITH THE WORK OF A

1533
01:19:19,622 --> 01:19:20,723
TALENTED PERSON, RAYANE DIBSY,

1534
01:19:20,723 --> 01:19:23,859
NOW IN POSTDOC TRAINING AT

1535
01:19:23,859 --> 01:19:24,460
PASTEUR INSTITUTE.

1536
01:19:24,460 --> 01:19:28,797
IF YOU IMAGINE THIS PATHWAY IN

1537
01:19:28,797 --> 01:19:39,341
THE CELL, VERY USEFUL, IF GAG

1538
01:19:39,341 --> 01:19:41,410
INTERACT WITH P53 YOU WILL

1539
01:19:41,410 --> 01:19:46,415
INTERFERE WITH THIS PATHWAY,

1540
01:19:46,415 --> 01:19:49,852
THERE'S TWO POSSIBILITIES.

1541
01:19:49,852 --> 01:19:53,455
WE DON'T KNOW EXACTLY THE

1542
01:19:53,455 --> 01:19:54,857
CHOLINERGIC PATHWAY ACTIVATION,

1543
01:19:54,857 --> 01:19:58,894
BUT WE TRY MIMIC IF WE BLOCK THE

1544
01:19:58,894 --> 01:20:03,065
PATHWAY AT THE END OF THIS

1545
01:20:03,065 --> 01:20:04,600
SIGNALING, WHAT HAPPEN?

1546
01:20:04,600 --> 01:20:08,971
SO FOR THAT WE USE CK-666, THAT

1547
01:20:08,971 --> 01:20:14,009
BLOCK UP TO THREE COMPLEX. 

1548
01:20:14,009 --> 01:20:16,312
WHEN DO YOU THAT, IT IS LESS

1549
01:20:16,312 --> 01:20:16,512
THAN.

1550
01:20:16,512 --> 01:20:20,316
WHEN YOU DO THAT IN T CELL WE

1551
01:20:20,316 --> 01:20:25,721
SEE THAT AT CERTAIN POINT, WE

1552
01:20:25,721 --> 01:20:27,456
HAVE A STRONG INCREASE IN

1553
01:20:27,456 --> 01:20:35,230
RELEASE, SAYING IF YOU HAVE LESS

1554
01:20:35,230 --> 01:20:36,765
DENSE ACTING, AND ANOTHER DRUG

1555
01:20:36,765 --> 01:20:38,867
WOULD NOT HAVE THIS EFFECT,

1556
01:20:38,867 --> 01:20:41,136
TRIED TO CHARACTERIZE THAT.

1557
01:20:41,136 --> 01:20:44,873
SO WE SHOW THAT JURKAT T CELL,

1558
01:20:44,873 --> 01:20:55,517
PRIMARY T CELL, EACH TIME HAVE

1559
01:20:55,517 --> 01:20:58,787
INCREASE. 

1560
01:20:58,787 --> 01:21:03,025
WE HAVE INCREASING RELEASE. 

1561
01:21:03,025 --> 01:21:06,662
AND THEN TRIED TO SEE IF IT WAS

1562
01:21:06,662 --> 01:21:10,466
INCREASE IN THE PLATFORM, SO WE

1563
01:21:10,466 --> 01:21:16,071
APPLIED THE SAME STORM/TIRF

1564
01:21:16,071 --> 01:21:17,372
MICROSCOPY DEVICE.

1565
01:21:17,372 --> 01:21:26,582
FIRST YOU CAN SEE T CELL HAVE

1566
01:21:26,582 --> 01:21:28,650
LESS DENSE ACTIN.

1567
01:21:28,650 --> 01:21:30,285
YOU CAN SEE THE CLUSTER

1568
01:21:30,285 --> 01:21:34,123
INDIVIDUAL PLATFORM WE CAN SEE

1569
01:21:34,123 --> 01:21:36,692
THAT WE HAVE INCREASE IN NUMBER

1570
01:21:36,692 --> 01:21:38,093
OF PLATFORM.

1571
01:21:38,093 --> 01:21:45,300
AND THEN IF WE RELATE TO THE

1572
01:21:45,300 --> 01:21:47,369
MESH WORK OF ACTIN WE HAVE A

1573
01:21:47,369 --> 01:21:52,508
NUMBER OF CLUSTER FROM ASSEMBLY

1574
01:21:52,508 --> 01:21:58,447
INCREASE FOR NUMBER OF ERROR.

1575
01:21:58,447 --> 01:22:01,183
GAG CAN ACCESS BETTER P2, BETTER

1576
01:22:01,183 --> 01:22:04,019
MEMBRANE, YOU CAN HAVE BETTER

1577
01:22:04,019 --> 01:22:04,553
ASSEMBLY.

1578
01:22:04,553 --> 01:22:08,857
WE TRY TO MIMIC THAT IN VITRO. 

1579
01:22:08,857 --> 01:22:14,463
SO THAT WE TOOK OUR BIOLIMITED

1580
01:22:14,463 --> 01:22:18,267
MEMBRANE WITH M1 AND PUT

1581
01:22:18,267 --> 01:22:19,501
DIFFERENT ACTING MESHWORK

1582
01:22:19,501 --> 01:22:21,003
DENSITY OF SURFACE OF MEMBRANE

1583
01:22:21,003 --> 01:22:23,238
AND LOOK AT P2 DEFICIENT.

1584
01:22:23,238 --> 01:22:28,043
WHAT WE OBSERVE P2 DEFICIENT WAS

1585
01:22:28,043 --> 01:22:34,082
DECREASING, AND WHAT HAPPENS

1586
01:22:34,082 --> 01:22:34,283
AGAIN.

1587
01:22:34,283 --> 01:22:37,352
IF WE DECREASE THE DYNAMIC IN

1588
01:22:37,352 --> 01:22:39,421
THE MEMBRANE GAG DIFFUSION DROPS

1589
01:22:39,421 --> 01:22:45,227
SO IF YOU HAVE LESS ACTIN YOU

1590
01:22:45,227 --> 01:22:48,297
HAVE MORE ACCESS TO P2

1591
01:22:48,297 --> 01:22:50,265
DIFFUSION, NUCLEATION OF GAG TO

1592
01:22:50,265 --> 01:22:54,636
MAKE A PARTICULAR ASSEMBLY WOULD

1593
01:22:54,636 --> 01:22:55,637
REQUIRE THAT.

1594
01:22:55,637 --> 01:23:04,813
SO THEN WE WANTED TO KNOW IF ANY

1595
01:23:04,813 --> 01:23:06,014
CELLULAR INHIBITOR OF ARP2/3

1596
01:23:06,014 --> 01:23:07,683
WOULD BE INVOLVED, WE TRIED SOME

1597
01:23:07,683 --> 01:23:16,091
AND COULD SEE HERE THAT WE HAD

1598
01:23:16,091 --> 01:23:21,029
GAG WITH SOME, ONE WAS ARPIN,

1599
01:23:21,029 --> 01:23:23,532
AND WE CAN SEE THIS COMPUTATION

1600
01:23:23,532 --> 01:23:28,337
WAS NOT EXISTING IF WE TAKE A

1601
01:23:28,337 --> 01:23:31,840
MINUS GAG NOT BINDING MEMBRANE

1602
01:23:31,840 --> 01:23:33,375
SO WE BELIEVE ARPIN WOULD BE

1603
01:23:33,375 --> 01:23:35,244
RECRUITED BY THE GAG PROTEIN OR

1604
01:23:35,244 --> 01:23:38,213
TRAPPED BY THE GAG PROTEIN, IN

1605
01:23:38,213 --> 01:23:42,918
ORDER TO HAVE LESS BRANCH

1606
01:23:42,918 --> 01:23:43,619
ACTIVITY.

1607
01:23:43,619 --> 01:23:45,320
SO WE CONFUSE THIS

1608
01:23:45,320 --> 01:23:46,955
IMMUNOPRECIPITATION BETWEEN GAG

1609
01:23:46,955 --> 01:23:52,761
AND ARPIN.

1610
01:23:52,761 --> 01:24:00,202
WHEN WE LOWER EXPRESSION, WE

1611
01:24:00,202 --> 01:24:02,070
LOWER HIV PARTICULAR, AND

1612
01:24:02,070 --> 01:24:04,573
FINALLY WE COULD SEE THAT WHEN

1613
01:24:04,573 --> 01:24:08,744
GAG IS EXPRESSED IN T CELL WE

1614
01:24:08,744 --> 01:24:11,013
INCREASE ARPIN IN THE MEMBRANE

1615
01:24:11,013 --> 01:24:13,215
FACTION, AND WE INCREASE ARPIN

1616
01:24:13,215 --> 01:24:15,517
AT THE T CELL SURFACE, AS YOU

1617
01:24:15,517 --> 01:24:19,254
CAN SEE HERE BY MICROSCOPY.

1618
01:24:19,254 --> 01:24:21,223
WE THINK THAT GAG IS ABOUT TO

1619
01:24:21,223 --> 01:24:24,493
ACTIVATE SOME OF THE ACTIN

1620
01:24:24,493 --> 01:24:28,330
PATHWAYS TO INHIBIT THE MESHWORK

1621
01:24:28,330 --> 01:24:31,199
TO HAVE MORE PARTICULAR

1622
01:24:31,199 --> 01:24:31,466
ASSEMBLY.

1623
01:24:31,466 --> 01:24:40,842
SO NOW WE WANT TO KNOW WHAT ARE

1624
01:24:40,842 --> 01:24:46,281
THE RECRUITMENT INVOLVING

1625
01:24:46,281 --> 01:24:46,982
POLYMERIZATION, THE

1626
01:24:46,982 --> 01:24:52,988
SPATIOCOMPONENTS, AND HOPEFULLY

1627
01:24:52,988 --> 01:24:55,524
NEXT YEAR.

1628
01:24:55,524 --> 01:24:56,758
HERE IS CONCLUSION, CONVINCE YOU

1629
01:24:56,758 --> 01:25:01,630
MANY OF US IN THE ROOM THAT WE

1630
01:25:01,630 --> 01:25:04,366
SHARE THE INTERESTS IN

1631
01:25:04,366 --> 01:25:08,203
UNDERSTANDING THE LIPID AND ML

1632
01:25:08,203 --> 01:25:09,871
RELATIONSHIP TO MAKE PARTICLES

1633
01:25:09,871 --> 01:25:11,707
OF THE CELL MEMBRANE IN

1634
01:25:11,707 --> 01:25:15,877
DIFFERENT CELL TYPE, DIFFERENT

1635
01:25:15,877 --> 01:25:17,179
SYSTEM, DIFFERENT GAG.

1636
01:25:17,179 --> 01:25:21,016
I WANT TO THANK MY TEAM BECAUSE

1637
01:25:21,016 --> 01:25:22,117
WITHOUT THEM NOTHING IS

1638
01:25:22,117 --> 01:25:25,821
POSSIBLE, BECAUSE I WISH I COULD

1639
01:25:25,821 --> 01:25:28,557
STILL DO EXPERIMENTS.

1640
01:25:28,557 --> 01:25:37,432
BUT ACTUALLY THE LAST EXPERIMENT

1641
01:25:37,432 --> 01:25:39,167
WAS THE MICROSCOPY, WE HAVE MORE

1642
01:25:39,167 --> 01:25:43,338
STUDENTS AT THAT TIME, SO WE DID

1643
01:25:43,338 --> 01:25:44,439
ALL THE EXPERIMENTS.

1644
01:25:44,439 --> 01:25:45,974
BUT TEN YEARS AGO NOW.

1645
01:25:45,974 --> 01:25:46,808
EIGHT YEARS AGO.

1646
01:25:46,808 --> 01:25:52,314
I WANT TO THANK THE SPONSORS AND

1647
01:25:52,314 --> 01:25:54,816
PLEASE COME AND VISIT

1648
01:25:54,816 --> 01:25:58,887
MONTPELLIER, A CITY BY THE SEA.

1649
01:25:58,887 --> 01:26:02,591
I WANT TO FINISH TO THANK ALAN

1650
01:26:02,591 --> 01:26:06,528
FOR ALL HIS YEARS OF EXCELLENT

1651
01:26:06,528 --> 01:26:09,698
SCIENCE, FOR FRIENDSHIP, AND

1652
01:26:09,698 --> 01:26:11,066
NON-SCIENTIFIC DISCUSSION ON

1653
01:26:11,066 --> 01:26:14,836
MUSIC, ART, AND THIS WAS A

1654
01:26:14,836 --> 01:26:18,106
SLIDE, ALSO I REMIND YOU OF

1655
01:26:18,106 --> 01:26:20,308
THIS.

1656
01:26:20,308 --> 01:26:26,348
 

1657
01:26:26,348 --> 01:26:33,255
 

1658
01:26:33,255 --> 01:26:34,689
YES, MARK.

1659
01:26:34,689 --> 01:26:39,494
>> ALL THIS WAY FROM

1660
01:26:39,494 --> 01:26:40,595
MONTPELLIER.

1661
01:26:40,595 --> 01:26:45,634
YOU MAY GO BACK TO (INAUDIBLE).

1662
01:26:45,634 --> 01:26:53,041
>> I KNOW YOU TRY TO
CONVINCE

1663
01:26:53,041 --> 01:26:53,809
ME.

1664
01:26:53,809 --> 01:26:57,646
>> ALL THE GAMMA
RETROVIRUSES IN

1665
01:26:57,646 --> 01:27:00,615
THE HUMAN GENOME INCLUDING GAG

1666
01:27:00,615 --> 01:27:03,552
PROTEIN HAVE NOW HUGE TREND ON

1667
01:27:03,552 --> 01:27:07,522
HOW GAG IS INVOLVED IN THE

1668
01:27:07,522 --> 01:27:12,694
FORMATION OF VESICLES.

1669
01:27:12,694 --> 01:27:13,428
>> YEAH.

1670
01:27:13,428 --> 01:27:14,429
>> THAT'S WONDERFUL.

1671
01:27:14,429 --> 01:27:18,433
>> THE OTHER ONE, SO AS

1672
01:27:18,433 --> 01:27:23,405
CHRISTINE -- (INAUDIBLE) ALL

1673
01:27:23,405 --> 01:27:27,776
THOSE RECEPTORS USED BY

1674
01:27:27,776 --> 01:27:29,811
ENVELOPES, LET'S SAY

1675
01:27:29,811 --> 01:27:30,512
TRANSPORTERS, REGULATE PATHWAYS,

1676
01:27:30,512 --> 01:27:35,650
I THINK THERE'S A WAY TO

1677
01:27:35,650 --> 01:27:37,753
ASSEMBLE IN THE HUMAN CELL ON

1678
01:27:37,753 --> 01:27:42,758
THE ONE HAND THE EQUIVALENT OF

1679
01:27:42,758 --> 01:27:47,362
ENVELOPES THAT WOULD BIND

1680
01:27:47,362 --> 01:27:50,198
TRANSPORTERS AND GAG, THE GAG

1681
01:27:50,198 --> 01:27:50,465
FORMATION.

1682
01:27:50,465 --> 01:27:51,633
>> YES, THAT'S ONE SIDE.

1683
01:27:51,633 --> 01:27:57,205
THE OTHER SIDE I WOULD SAY,

1684
01:27:57,205 --> 01:28:02,577
WE'RE SHOWING CERTAIN

1685
01:28:02,577 --> 01:28:03,678
POPULATIONS OF EXTRACELLULAR

1686
01:28:03,678 --> 01:28:14,089
VESICLES, AND THERE'S A

1687
01:28:17,959 --> 01:28:25,000
BEAUTIFUL WORK, THEY ARE

1688
01:28:25,000 --> 01:28:25,333
INTERACTING.

1689
01:28:25,333 --> 01:28:27,536
TO SEND DISTANT SIGNAL FOR

1690
01:28:27,536 --> 01:28:32,007
MEMORY, SO, THIS IS, YOU KNOW, A

1691
01:28:32,007 --> 01:28:34,309
VERY OPEN PERIOD.

1692
01:28:34,309 --> 01:28:36,511
I LIKE THE IDEA OF RETROVIRUSES

1693
01:28:36,511 --> 01:28:39,881
USING THAT PATHWAY TO GET OUT

1694
01:28:39,881 --> 01:28:44,186
AND TO SEND A SIGNAL,

1695
01:28:44,186 --> 01:28:54,729
TRANSPORTATION, TO LONG DISTANCE

1696
01:29:13,715 --> 01:29:23,892
IN FORMATION. 

1697
01:29:29,264 --> 01:29:32,133
>> NEXT SPEAKER IS RIGHT
NOW IN

1698
01:29:32,133 --> 01:29:33,335
URUGUAY, I BELIEVE.

1699
01:29:33,335 --> 01:29:37,172
HE WAS SUPPOSED TO BE HERE BUT

1700
01:29:37,172 --> 01:29:40,208
UNITED AIRLINES DIDN'T

1701
01:29:40,208 --> 01:29:40,475
COOPERATE.

1702
01:29:40,475 --> 01:29:45,146
NO, SORRY, SORRY.

1703
01:29:45,146 --> 01:29:51,853
NEXT SPEAKER IS HERE.

1704
01:29:51,853 --> 01:29:52,520
IT'S KARIN MUSIER-FORSYTH, A

1705
01:29:52,520 --> 01:29:56,558
NAME FAMILIAR TO MOST OF US I'M

1706
01:29:56,558 --> 01:29:58,693
SURE. 

1707
01:29:58,693 --> 01:30:04,566
I WILL JUST SAY THAT KARIN WAS

1708
01:30:04,566 --> 01:30:06,401
TRAINED IN tRNA BIOCHEMISTRY

1709
01:30:06,401 --> 01:30:10,338
AND INTERACTION OF tRNAs

1710
01:30:10,338 --> 01:30:13,074
WITH CELLULAR PROTEINS.

1711
01:30:13,074 --> 01:30:16,378
AND I FIRST BECAME AWARE WITH

1712
01:30:16,378 --> 01:30:17,012
HER FIRST RETROVIRUS PAPER, I

1713
01:30:17,012 --> 01:30:24,586
THINK IT WAS THE FIRST

1714
01:30:24,586 --> 01:30:25,553
RETROVIRUS PAPER, AND IT STRUCK

1715
01:30:25,553 --> 01:30:28,523
ME FROM THE FIRST PAPER HERE IS

1716
01:30:28,523 --> 01:30:32,994
SOMEONE WHO IS REALLY INTERESTED

1717
01:30:32,994 --> 01:30:36,631
IN HOW THE VIRAL PROTEINS ARE

1718
01:30:36,631 --> 01:30:40,101
MANIPULATING THE RNAs,

1719
01:30:40,101 --> 01:30:43,838
INTERACTING WITH RNAs, IT WAS

1720
01:30:43,838 --> 01:30:46,274
A FASCINATING PAPER, AND SHE'S

1721
01:30:46,274 --> 01:30:49,444
CONTINUED TO DO FASCINATING

1722
01:30:49,444 --> 01:30:50,879
WORK, EVER SINCE.

1723
01:30:50,879 --> 01:30:52,080
WE'VE REALLY KNOWN EACH OTHER

1724
01:30:52,080 --> 01:30:54,916
QUITE A WHILE NOW, HAD A NUMBER

1725
01:30:54,916 --> 01:30:55,917
OF PAPERS TOGETHER.

1726
01:30:55,917 --> 01:31:04,559
AND TODAY SHE'S GOING TO TELL US

1727
01:31:04,559 --> 01:31:06,594
ABOUT WORK, REALLY AMAZING WORK.

1728
01:31:06,594 --> 01:31:09,597
WITHOUT FURTHER ADO, THANK YOU

1729
01:31:09,597 --> 01:31:10,799
FOR COMING, KARIN, AND THANK YOU

1730
01:31:10,799 --> 01:31:20,976
FOR BEING PART OF THIS EVENT. 

1731
01:31:20,976 --> 01:31:21,977
 

1732
01:31:21,977 --> 01:31:23,712
>> IT'S AN HONOR AND
PLEASURE TO

1733
01:31:23,712 --> 01:31:26,214
BE HERE TO CELEBRATE ALAN'S LONG

1734
01:31:26,214 --> 01:31:27,315
CAREER.

1735
01:31:27,315 --> 01:31:31,152
WE DID MEET IN 1997, MY FIRST

1736
01:31:31,152 --> 01:31:32,687
COLD SPRING HARBOR MEETING.

1737
01:31:32,687 --> 01:31:33,888
AND BASICALLY WE'VE BEEN TALKING

1738
01:31:33,888 --> 01:31:34,856
EVER SINCE.

1739
01:31:34,856 --> 01:31:40,228
AT THAT MEETING I ALSO MET

1740
01:31:40,228 --> 01:31:41,329
WEI-SHAU, STEVE AND EVERYBODY

1741
01:31:41,329 --> 01:31:43,198
HERE, A LOT OF PEOPLE HERE.

1742
01:31:43,198 --> 01:31:46,034
TWO YEARS LATER HAD A

1743
01:31:46,034 --> 01:31:46,334
SABBATICAL.

1744
01:31:46,334 --> 01:31:49,871
I DECIDED TO DO IT HERE AND BE

1745
01:31:49,871 --> 01:31:50,739
IMMERSED IN RETROVIROLOGY.

1746
01:31:50,739 --> 01:31:52,173
THAT'S WHAT I DID.

1747
01:31:52,173 --> 01:31:54,476
AND SO I THINK IT WAS THE FALL

1748
01:31:54,476 --> 01:31:57,312
OF 1999, I SPENT FIVE MONTHS

1749
01:31:57,312 --> 01:31:58,413
HERE, INTERACTED, MANY MEALS

1750
01:31:58,413 --> 01:32:01,249
WITH ALAN, LOTS OF CONVERSATIONS

1751
01:32:01,249 --> 01:32:05,320
WITH  ALAN AND CONTINUED TO THIS

1752
01:32:05,320 --> 01:32:05,520
DAY.

1753
01:32:05,520 --> 01:32:09,791
I'M HAPPY TO TELL YOU ABOUT

1754
01:32:09,791 --> 01:32:11,459
COLLABORATION WITH WEI-SHU AND

1755
01:32:11,459 --> 01:32:15,597
HER TEAM, MY TEAM IS HERE ON THE

1756
01:32:15,597 --> 01:32:17,232
LEFT.

1757
01:32:17,232 --> 01:32:19,868
MIKE IS ASSISTANT PROFESSOR, AND

1758
01:32:19,868 --> 01:32:22,170
JONATHAN WAS A TRAINEE, Ph.D.

1759
01:32:22,170 --> 01:32:26,207
WITH ME, NOW POSTDOC WITH

1760
01:32:26,207 --> 01:32:26,775
WEI-SHAU.

1761
01:32:26,775 --> 01:32:28,843
HOW DO I ADVANCE SLIDES?

1762
01:32:28,843 --> 01:32:29,144
OKAY.

1763
01:32:29,144 --> 01:32:34,082
WE ALL KNOW ALL THE CURRENT HIV

1764
01:32:34,082 --> 01:32:35,617
THERAPEUTICS ACT MAINLY ON

1765
01:32:35,617 --> 01:32:37,485
ENZYMES, WE'RE INTERESTED IN

1766
01:32:37,485 --> 01:32:45,026
EXPANDING THE RANGE OF DRUGGABLE

1767
01:32:45,026 --> 01:32:46,461
TARGETS TO RNA.

1768
01:32:46,461 --> 01:32:49,731
IT'S A TEMPLATE FOR TRANSLATION,

1769
01:32:49,731 --> 01:32:55,336
GAG AND GAGPOL, AND GENOME, SO

1770
01:32:55,336 --> 01:32:56,538
PRESUMABLY IDENTICAL MOLECULE IS

1771
01:32:56,538 --> 01:32:57,839
PACKAGED AS A DIMER.

1772
01:32:57,839 --> 01:33:00,742
WHAT'S CLEAR IN RECENT YEARS IS

1773
01:33:00,742 --> 01:33:02,110
THAT THIS DIFFERENCE IN THE

1774
01:33:02,110 --> 01:33:04,212
STATE OF THE FULL LENGTH

1775
01:33:04,212 --> 01:33:05,947
INSPLICED RNA IS DETERMINED AT

1776
01:33:05,947 --> 01:33:07,148
THE LEVEL OF TRANSCRIPTION.

1777
01:33:07,148 --> 01:33:11,419
AND SO IF YOU LOOK AT WHAT HOST

1778
01:33:11,419 --> 01:33:11,986
RNA POLYMERASE TRANSCRIBED

1779
01:33:11,986 --> 01:33:15,590
YOU'LL SEE THERE'S A MIXTURE OF

1780
01:33:15,590 --> 01:33:17,459
SPECIES, THE VIRUS ENCODES 3G,

1781
01:33:17,459 --> 01:33:20,628
YOU GET 3Gs IN A CAP, TWO,

1782
01:33:20,628 --> 01:33:22,063
ONE, A BUNCH OF MISCELLANEOUS

1783
01:33:22,063 --> 01:33:23,932
OTHER THINGS.

1784
01:33:23,932 --> 01:33:26,668
A NOBODY OF LABS HAVE SHOWN IF

1785
01:33:26,668 --> 01:33:29,404
YOU LOOK IN THE CELL THAT MOST

1786
01:33:29,404 --> 01:33:31,840
OF THE RNA IS 3G AS YOU WOULD

1787
01:33:31,840 --> 01:33:33,808
PREDICT BUT THERE'S SIGNIFICANT

1788
01:33:33,808 --> 01:33:35,910
AMOUNT OF 2G, 1G, OTHER SPECIES

1789
01:33:35,910 --> 01:33:36,678
AS WELL.

1790
01:33:36,678 --> 01:33:38,413
HOWEVER, IF YOU LOOK IN THE

1791
01:33:38,413 --> 01:33:41,249
VIRUS PARTICLE, WHAT YOU SEE IS

1792
01:33:41,249 --> 01:33:45,420
IT'S NOW ALMOST -- PREDOMINANTLY

1793
01:33:45,420 --> 01:33:48,490
1G WITH A LITTLE BIT OF 3G,

1794
01:33:48,490 --> 01:33:49,290
SELECTED MECHANISM OF PACKAGING

1795
01:33:49,290 --> 01:33:50,325
GOING ON HERE.

1796
01:33:50,325 --> 01:33:52,894
THAT'S THE BASIS OF THE PROJECT.

1797
01:33:52,894 --> 01:34:00,768
HOW ARE THE 1 Gs PACKAGES AND

1798
01:34:00,768 --> 01:34:02,036
ARE 3Gs PREFERENTIALLY

1799
01:34:02,036 --> 01:34:02,370
TRANSLATED.

1800
01:34:02,370 --> 01:34:04,472
I NEED TO INTRODUCE OR REMIND

1801
01:34:04,472 --> 01:34:07,308
YOU OF WHAT THE IMPORTANT RNA

1802
01:34:07,308 --> 01:34:09,177
STRUCTURAL ELEMENTS IN THE FIVE

1803
01:34:09,177 --> 01:34:11,779
PRIME INTRANSLATED REGION.

1804
01:34:11,779 --> 01:34:15,416
WE SAW THESE STRUCTURES, HERE IS

1805
01:34:15,416 --> 01:34:18,686
THE TAR/POLY-A, TAR LOOP, POLY-A

1806
01:34:18,686 --> 01:34:20,121
LOOP, PRIMER BINDING SITE

1807
01:34:20,121 --> 01:34:21,890
DOMAIN.

1808
01:34:21,890 --> 01:34:27,762
AND THEN THIS IS THE SITE

1809
01:34:27,762 --> 01:34:34,602
PACKAGING SIGNAL WITH THE

1810
01:34:34,602 --> 01:34:37,171
PALINDROMIC LOOP, IMPORTANT FOR

1811
01:34:37,171 --> 01:34:42,010
INITIATING DIMERIZATION, AND A

1812
01:34:42,010 --> 01:34:43,211
NUMBER OF SINGLE-STRANDED GENES

1813
01:34:43,211 --> 01:34:45,413
ARE IMPORTANT FOR BINDING.

1814
01:34:45,413 --> 01:34:46,948
WHAT I'LL FOCUS ON TODAY IS

1815
01:34:46,948 --> 01:34:51,119
WHERE YOU HAVE THE VARIABLE

1816
01:34:51,119 --> 01:34:53,755
NUMBER, HOW THEY DETERMINE THE

1817
01:34:53,755 --> 01:34:56,491
STATE OF THIS GENOMIC RNA.

1818
01:34:56,491 --> 01:35:00,628
AND AS MANY OF YOU KNOW, MANY

1819
01:35:00,628 --> 01:35:03,264
FIVE PRIME UNTRANSLATED REGIONS

1820
01:35:03,264 --> 01:35:05,233
OF HUMAN GENES, ALL KINDS OF

1821
01:35:05,233 --> 01:35:15,643
GENES, ARE REGULATED BY

1822
01:35:16,277 --> 01:35:17,512
SO-CALLED RIBAL STRICTURES, THIS

1823
01:35:17,512 --> 01:35:19,514
EXPRESSION WOULD OCCUR.

1824
01:35:19,514 --> 01:35:22,584
HOWEVER, IF A LIGAND BINDS THAT

1825
01:35:22,584 --> 01:35:23,818
CAN CHANGE THE CONFIRMATIONAL

1826
01:35:23,818 --> 01:35:26,521
STATE, TURNING THE GENE OFF.

1827
01:35:26,521 --> 01:35:28,056
SO WE KNEW ABOUT THIS PARADIGM

1828
01:35:28,056 --> 01:35:30,658
AND WHAT WE WERE WONDERING IS,

1829
01:35:30,658 --> 01:35:31,759
WELL, MAYBE IT'S NOT SMALL

1830
01:35:31,759 --> 01:35:33,294
MOLECULE BINDING BUT MAYBE IT'S

1831
01:35:33,294 --> 01:35:35,163
THE NUMBER OF GENES REGULATING

1832
01:35:35,163 --> 01:35:38,099
THE CONFIRMATION OF THIS RNA.

1833
01:35:38,099 --> 01:35:40,735
AND SO THAT IF YOU HAVE 3 Gs,

1834
01:35:40,735 --> 01:35:42,604
THIS IS A CONFIRMATION OF

1835
01:35:42,604 --> 01:35:45,006
PREFERENTIAL TRANSLATION, IF YOU

1836
01:35:45,006 --> 01:35:46,341
HAVE 1G, FORMS A CONFIRMATION

1837
01:35:46,341 --> 01:35:48,076
THAT'S PREFERENTIAL TO THE

1838
01:35:48,076 --> 01:35:48,943
PACKAGING.

1839
01:35:48,943 --> 01:35:51,879
SO TO LOOK AT THIS, THIS WAS

1840
01:35:51,879 --> 01:35:54,215
PUBLISHED A NUMBER OF YEARS AGO,

1841
01:35:54,215 --> 01:35:56,718
I'M GOING TO SUMMARIZE.

1842
01:35:56,718 --> 01:35:59,354
WE PERFORMED RNA STRUCTURE

1843
01:35:59,354 --> 01:36:01,189
PRONING TECHNIQUE CALLED SHAPE,

1844
01:36:01,189 --> 01:36:03,191
AND WE RAN -- FIRST OF ALL I CAN

1845
01:36:03,191 --> 01:36:05,493
SHOW WHEN YOU TAKE THE 1G RNA

1846
01:36:05,493 --> 01:36:07,996
AND RUN IT ON A NATIVE GEL, WHAT

1847
01:36:07,996 --> 01:36:10,632
YOU SEE RIGHT AWAY IS THAT IT'S

1848
01:36:10,632 --> 01:36:11,833
MOSTLY DIMERIC SPECIES WITH A

1849
01:36:11,833 --> 01:36:13,668
LITTLE BIT OF MONOMER, BUT IF

1850
01:36:13,668 --> 01:36:17,071
YOU TAKE THE EXACT SAME 3G RNA,

1851
01:36:17,071 --> 01:36:19,040
FOLD IT UNDER THE SAME

1852
01:36:19,040 --> 01:36:20,908
CONDITIONS, WE SEE MAINLY

1853
01:36:20,908 --> 01:36:21,676
MONOMERIC SPECIES.

1854
01:36:21,676 --> 01:36:25,146
AND SO WE CUT OUT THESE BANDS

1855
01:36:25,146 --> 01:36:27,382
THROUGH GEL AND ASSOCIATED IN

1856
01:36:27,382 --> 01:36:28,883
CHEMICAL REAGENTS THAT PROBE RNA

1857
01:36:28,883 --> 01:36:32,854
STRUCTURE AND WHAT WE FOUND WAS

1858
01:36:32,854 --> 01:36:35,790
THAT THE 1G RNA, PRIMARILY

1859
01:36:35,790 --> 01:36:37,458
DIMERIC, HAS THIS SECONDARY

1860
01:36:37,458 --> 01:36:38,526
STRUCTURE FAIRLY SIMILAR TO WHAT

1861
01:36:38,526 --> 01:36:40,628
I SHOWED EARLIER.

1862
01:36:40,628 --> 01:36:43,831
YOU CAN SEE THE TAR CONFORMS,

1863
01:36:43,831 --> 01:36:45,867
POLY-A CONFORMS, YOU GET INTO

1864
01:36:45,867 --> 01:36:51,105
THE PRIMER BINDING SITE.

1865
01:36:51,105 --> 01:36:53,441
I'M CIRCLING THESE, CRITICAL FOR

1866
01:36:53,441 --> 01:36:54,542
GAG BINDING, PACKAGING RNA,

1867
01:36:54,542 --> 01:36:57,712
EXPOSED AND AVAILABLE IN STINGEL

1868
01:36:57,712 --> 01:37:04,385
STRANDED REGIONS.

1869
01:37:04,385 --> 01:37:07,889
IF YOU TAKE THE 3G RNA, WE'VE

1870
01:37:07,889 --> 01:37:09,857
ADDED TWO MORE Gs.

1871
01:37:09,857 --> 01:37:12,260
WHAT YOU'LL SEE HAPPEN IN THE

1872
01:37:12,260 --> 01:37:14,662
NEXT SLIDE THAT NEXT G IS PAIRED

1873
01:37:14,662 --> 01:37:20,902
WITH C, AND WHAT YOU GET, WHAT

1874
01:37:20,902 --> 01:37:25,740
WAS SURPRISING, COMPLETE

1875
01:37:25,740 --> 01:37:29,444
REARRANGEMENT OF THE RNA,

1876
01:37:29,444 --> 01:37:32,613
STEALING NUCLEOTIDE FROM POLY-A,

1877
01:37:32,613 --> 01:37:34,248
RESULTING IN UNFOLDINGS, DOWN

1878
01:37:34,248 --> 01:37:35,583
EXTREME INTERACTION, FURTHER

1879
01:37:35,583 --> 01:37:36,984
DOWNSTREAM EFFECTS, NOW Gs

1880
01:37:36,984 --> 01:37:39,287
WHICH WERE CLOSE TOGETHER AND

1881
01:37:39,287 --> 01:37:40,922
SINGLE-STRANDED HERE ARE NOW

1882
01:37:40,922 --> 01:37:44,425
FURTHER APART AND PARTIALLY

1883
01:37:44,425 --> 01:37:44,726
SEQUESTERED.

1884
01:37:44,726 --> 01:37:45,960
AND DIMERIZATION INITIATION SITE

1885
01:37:45,960 --> 01:37:50,898
IS NO LONGER IN A PALINDROMIC

1886
01:37:50,898 --> 01:37:51,232
LOOP.

1887
01:37:51,232 --> 01:37:55,837
THIS HELPED TO EXPLAIN WE THINK

1888
01:37:55,837 --> 01:38:00,108
WHY THIS IS FOR PACKAGING, HAS

1889
01:38:00,108 --> 01:38:02,944
EXPOSED DIS, HAS THE G BINDING

1890
01:38:02,944 --> 01:38:04,378
SITES FOR PACKAGING EXPOSED, AND

1891
01:38:04,378 --> 01:38:08,082
WHAT WE DIDN'T HAVE PREDICTED

1892
01:38:08,082 --> 01:38:13,221
INTACT POLY-A HAIRPIN.

1893
01:38:13,221 --> 01:38:15,523
THOSE ELEMENTS ARE MISSING IN

1894
01:38:15,523 --> 01:38:20,361
THE 3G RNA NOT PREFERENTIALLY

1895
01:38:20,361 --> 01:38:20,628
PACKAGED.

1896
01:38:20,628 --> 01:38:22,563
AGAIN, WE IDENTIFIED TWO SETS OF

1897
01:38:22,563 --> 01:38:27,468
MUTANTS THAT ELIMINATED SELECTED

1898
01:38:27,468 --> 01:38:31,072
1G RNA PACKAGING.

1899
01:38:31,072 --> 01:38:36,911
WE DESTABILIZED BOND OF POLY-A.

1900
01:38:36,911 --> 01:38:37,779
USE ARTIFICIALLY DESTABILIZED,

1901
01:38:37,779 --> 01:38:40,615
YOU SEE IN CONTRAST TO WILD TYPE

1902
01:38:40,615 --> 01:38:42,483
WITH THIS DRAMATIC DIFFERENCE IN

1903
01:38:42,483 --> 01:38:45,853
STRUCTURE OF THE 3G AND 1G AND

1904
01:38:45,853 --> 01:38:46,854
PREFERENTIAL PACKAGING.

1905
01:38:46,854 --> 01:38:49,524
1G RNA NOW YOU ELIMINATE THAT

1906
01:38:49,524 --> 01:38:51,292
PREFERENTIAL PACKAGING AND WHEN

1907
01:38:51,292 --> 01:38:55,096
WE PROBE THE STRUCTURE FOR

1908
01:38:55,096 --> 01:38:56,964
EXAMPLE ON THE NATIVE GEL NO

1909
01:38:56,964 --> 01:39:02,003
MATTER IF YOU HAVE 3G OR 1G,

1910
01:39:02,003 --> 01:39:04,205
THEY ALL LOOK 3G LIKE.

1911
01:39:04,205 --> 01:39:06,707
WE'VE ELIMINATED THAT STRUCTURAL

1912
01:39:06,707 --> 01:39:08,442
DIFFERENCE AND ELIMINATED

1913
01:39:08,442 --> 01:39:10,111
SELECTIVE PACKAGING.

1914
01:39:10,111 --> 01:39:12,847
ANOTHER SET HAD TWO NUCLEOSIDES

1915
01:39:12,847 --> 01:39:19,086
INSERTED, THIS GENE TO BASE PAIR

1916
01:39:19,086 --> 01:39:21,823
WITH THIS, ANY TWO NUCLEOSIDES

1917
01:39:21,823 --> 01:39:24,325
ELIMINATED SELECTED 1G PACKAGING

1918
01:39:24,325 --> 01:39:26,294
IN CELLS AND ALSO ELIMINATED

1919
01:39:26,294 --> 01:39:27,395
STRUCTURAL DIFFERENCES BETWEEN

1920
01:39:27,395 --> 01:39:27,728
RNAs.

1921
01:39:27,728 --> 01:39:30,131
NOW THEY ALL LOOK LIKE THEY ARE

1922
01:39:30,131 --> 01:39:36,804
1G AND THERE'S NO PREFERENTIAL

1923
01:39:36,804 --> 01:39:37,905
PACKAGING.

1924
01:39:37,905 --> 01:39:41,075
WE FOUND THAT SELECTED 1G

1925
01:39:41,075 --> 01:39:46,547
PACKAGING IS THE CONFIRMATIONAL

1926
01:39:46,547 --> 01:39:48,282
ENSEMBLES THAT HAVE DIFFERENCES,

1927
01:39:48,282 --> 01:39:52,320
IN PARTICULAR DIFFERENCES IN

1928
01:39:52,320 --> 01:39:54,422
POLY-A HAIRPIN STABILITY.

1929
01:39:54,422 --> 01:39:55,957
YOU HAVE DIS EXPOSURE, EXPOSED

1930
01:39:55,957 --> 01:39:58,893
GAG BINDING SITES, AND

1931
01:39:58,893 --> 01:40:03,931
PACKAGING, BUT IF THE POLY-I

1932
01:40:03,931 --> 01:40:05,032
HAIRPIN IS DISRUPTED

1933
01:40:05,032 --> 01:40:06,367
ARTIFICIALLY OR THROUGH

1934
01:40:06,367 --> 01:40:11,405
INTERACTION THE DIS IS

1935
01:40:11,405 --> 01:40:18,613
SEQUESTERED, DNA BINDING SITES

1936
01:40:18,613 --> 01:40:19,614
ARE SEQUESTERED.

1937
01:40:19,614 --> 01:40:21,015
GAG SHOULD BIND AND ASSOCIATE

1938
01:40:21,015 --> 01:40:24,318
WITH THE 1G RNA WHICH HAS

1939
01:40:24,318 --> 01:40:26,254
EXPOSED GAG BINDING SITE.

1940
01:40:26,254 --> 01:40:28,689
WE'VE TRIED HARD TO LOOK AT

1941
01:40:28,689 --> 01:40:29,991
DIFFERENCES IN AFFINITY, WE

1942
01:40:29,991 --> 01:40:37,431
DON'T SEE ANY DIFFERENCES..

1943
01:40:37,431 --> 01:40:40,067
GANG IS PROMISCUOUS BINDING

1944
01:40:40,067 --> 01:40:41,168
PROTEIN, HAS PREFERENCES TO

1945
01:40:41,168 --> 01:40:42,603
SINGLE-STRANDED GENE BUT IF WE

1946
01:40:42,603 --> 01:40:44,105
LOOK AT BINDING AFFINITY WE

1947
01:40:44,105 --> 01:40:45,439
DON'T SEE DIFFERENCES.

1948
01:40:45,439 --> 01:40:48,709
WHAT WE DECIDED TO DO, WE

1949
01:40:48,709 --> 01:40:50,778
PREDICT THAT GAG WOULD ASSEMBLE

1950
01:40:50,778 --> 01:40:56,384
MORE READILY ONTO THIS RNA AND

1951
01:40:56,384 --> 01:41:03,491
SO WE USED MASS PHOTOMETRY, A

1952
01:41:03,491 --> 01:41:04,392
SOLUTION-BASED MASS MEASUREMENT

1953
01:41:04,392 --> 01:41:06,894
TECHNIQUE, YOU HAVE YOUR

1954
01:41:06,894 --> 01:41:11,465
MOLECULES IN SOLUTION, SHINE

1955
01:41:11,465 --> 01:41:14,001
INCIDENT LIGHT ON THE MOLECULE.

1956
01:41:14,001 --> 01:41:15,770
THEY WILL SCATTER THE LIGHT AS

1957
01:41:15,770 --> 01:41:17,271
THEY HIT THE GLASS SURFACE SO

1958
01:41:17,271 --> 01:41:20,041
YOU CAN MEASURE THAT SCATTERING.

1959
01:41:20,041 --> 01:41:23,744
THIS IS ALL CALIBRATED SUCH THAT

1960
01:41:23,744 --> 01:41:26,047
THE SCATTERED LIGHT IS BASICALLY

1961
01:41:26,047 --> 01:41:30,618
IN RELATION TO THE MOLECULE AND

1962
01:41:30,618 --> 01:41:33,354
CALIBRATED WITH PROTEIN RNA

1963
01:41:33,354 --> 01:41:33,621
STANDARDS.

1964
01:41:33,621 --> 01:41:35,756
YOU TAKE MOVIES, BUILD UP

1965
01:41:35,756 --> 01:41:37,725
HISTOGRAMS, CAN SEE DIFFERENT

1966
01:41:37,725 --> 01:41:39,827
SIZES OF COMPLEXES FORMING.

1967
01:41:39,827 --> 01:41:43,631
WE USED OUR 1G AND 3G RNAs,

1968
01:41:43,631 --> 01:41:46,701
WHICH WE MIMICKED BY PUTTING 2G

1969
01:41:46,701 --> 01:41:48,235
AND 4G, I FORGOT TO SAY THAT.

1970
01:41:48,235 --> 01:41:55,910
AND THAT'S WHY THIS IS LABELED

1971
01:41:55,910 --> 01:41:58,079
2G, 4G, OVERLATE MASS

1972
01:41:58,079 --> 01:42:00,615
SPECTROMETRY DATA HERE, THE

1973
01:42:00,615 --> 01:42:02,783
ORANGE -- HERE WE HAVE THE COUNT

1974
01:42:02,783 --> 01:42:05,186
VERSUS MASS, EACH PEAK IS A

1975
01:42:05,186 --> 01:42:05,953
POPULATION OF MOLECULES WITH

1976
01:42:05,953 --> 01:42:06,954
THIS MASS.

1977
01:42:06,954 --> 01:42:10,124
WHAT YOU CAN SEE IS THAT IN A

1978
01:42:10,124 --> 01:42:14,595
CASE OF THE 4G, WE SEE THE RNA

1979
01:42:14,595 --> 01:42:16,497
MONOMER, RNA DIMER, AND WE ADD

1980
01:42:16,497 --> 01:42:19,100
GAG, WE ONLY SEE A LITTLE BIT OF

1981
01:42:19,100 --> 01:42:20,001
COMPLEX FORMATION.

1982
01:42:20,001 --> 01:42:21,202
BUT UNDER THE EXACT SAME

1983
01:42:21,202 --> 01:42:23,270
CONDITIONS WHEN WE LOOK AT GAG

1984
01:42:23,270 --> 01:42:25,473
INTERACTING WITH THE 2G RNA, WE

1985
01:42:25,473 --> 01:42:29,076
AGAIN SEE THE MONOMERIC AND

1986
01:42:29,076 --> 01:42:30,511
DIMERIC RNA BUT SEE BUILDUP OF

1987
01:42:30,511 --> 01:42:36,317
GAG ON RNA WHERE WE CAN MEASURE

1988
01:42:36,317 --> 01:42:39,153
1G,  2G, 3G, 4G, 5G, 6G AND

1989
01:42:39,153 --> 01:42:42,223
HIGHER ORDERS OF COMPLEXES. 

1990
01:42:42,223 --> 01:42:46,260
THIS SHOWS GAG ASSEMBLES MORE

1991
01:42:46,260 --> 01:42:47,361
FAVORABLY ON 1G.

1992
01:42:47,361 --> 01:42:49,530
THE OTHER PREDICTION FROM THIS

1993
01:42:49,530 --> 01:42:53,701
EXPERIMENT IS THAT YOU -- WE SEE

1994
01:42:53,701 --> 01:42:57,238
DIFFERENCES IN POLY-A HAIRPIN

1995
01:42:57,238 --> 01:42:58,839
STABILITY APPEAR TO IMPACT

1996
01:42:58,839 --> 01:42:59,940
PACKAGING.

1997
01:42:59,940 --> 01:43:03,677
WE WANTED TO MEASURE POLY-A

1998
01:43:03,677 --> 01:43:05,112
HAIRPIN STABILITY.

1999
01:43:05,112 --> 01:43:07,948
WE USED THERMAL MELTING

2000
01:43:07,948 --> 01:43:08,916
EXPERIMENTS, WHERE WE JUST

2001
01:43:08,916 --> 01:43:10,684
LOOKED, IF YOU'RE FAMILIAR WITH

2002
01:43:10,684 --> 01:43:12,853
THESE, YOU CAN LOOK AT THE

2003
01:43:12,853 --> 01:43:15,923
ABSORBANCE AS A FUNCTION OF

2004
01:43:15,923 --> 01:43:17,591
TEMPERATURE AND GET THESE

2005
01:43:17,591 --> 01:43:18,793
CURVES, MID-POINT IS MELTING

2006
01:43:18,793 --> 01:43:20,528
TEMPERATURE, YOU CAN TAKE THE

2007
01:43:20,528 --> 01:43:21,762
FIRST DERIVATIVE OF THIS AND IN

2008
01:43:21,762 --> 01:43:23,564
THIS CASE THE PEAK IS THE

2009
01:43:23,564 --> 01:43:24,398
MELTING TEMPERATURE.

2010
01:43:24,398 --> 01:43:27,001
I'LL BE SHOWING YOU SHE'S TYPES

2011
01:43:27,001 --> 01:43:27,868
OF CURVES.

2012
01:43:27,868 --> 01:43:31,405
AND THIS IS DONE BY MICHAEL IN

2013
01:43:31,405 --> 01:43:32,606
MY LAB, UNDERGRADUATE.

2014
01:43:32,606 --> 01:43:39,180
WHAT HE LOOKED AT WAS THEY ARE

2015
01:43:39,180 --> 01:43:40,347
-- THERMAL STABILITY OF TAR.

2016
01:43:40,347 --> 01:43:42,550
IF YOU USE THE PROGRAM LIKE RNA

2017
01:43:42,550 --> 01:43:46,253
STRUCTURE TO PREDICT THE

2018
01:43:46,253 --> 01:43:47,588
DIFFERENCES IN STABILITY

2019
01:43:47,588 --> 01:43:50,891
PRE-DISTRICT THE -- PREDICT THE

2020
01:43:50,891 --> 01:43:53,394
ENERGY OF THE 4G IS MORE STABLE.

2021
01:43:53,394 --> 01:43:55,796
WE TAKE THE FIRST DERIVATIVE,

2022
01:43:55,796 --> 01:43:58,099
SLIGHT INCREASE IN STABILITY FOR

2023
01:43:58,099 --> 01:44:01,502
THE 4G WHICH HAS THAT GC PAIR.

2024
01:44:01,502 --> 01:44:07,741
WANTED TO LOOK AT TAR POLY-A,

2025
01:44:07,741 --> 01:44:11,245
HERE IS TAR.

2026
01:44:11,245 --> 01:44:13,147
POLY-A HAIRPIN, 2G AND 4G

2027
01:44:13,147 --> 01:44:13,547
VERSIONS.

2028
01:44:13,547 --> 01:44:16,016
IF YOU USE RNA STRUCTURES IT

2029
01:44:16,016 --> 01:44:17,451
DEPREDICTS IF THESE ARE EXACTLY

2030
01:44:17,451 --> 01:44:18,519
THE SAME IN STABILITY.

2031
01:44:18,519 --> 01:44:20,387
IF YOU DO THE MEASUREMENT,

2032
01:44:20,387 --> 01:44:21,922
THAT'S NOT ALL YOU SEE.

2033
01:44:21,922 --> 01:44:25,626
WE SEE A MUCH MORE STABLE

2034
01:44:25,626 --> 01:44:27,061
OVERALL PROFILE FOR 2G, AND IF

2035
01:44:27,061 --> 01:44:29,797
YOU TAKE THE DERIVATIVE YOU CAN

2036
01:44:29,797 --> 01:44:32,766
TEASE OUT DIFFERENT MELTING

2037
01:44:32,766 --> 01:44:33,868
TEMPERATURES.

2038
01:44:33,868 --> 01:44:36,270
WE SEE TWO MELTING TEMPERATURES,

2039
01:44:36,270 --> 01:44:40,407
ONE FOR TAR, ONE FOR POLY-A.

2040
01:44:40,407 --> 01:44:43,144
AGAIN THE TAR IS MORE STABLE FOR

2041
01:44:43,144 --> 01:44:44,545
THE 4G RNA.

2042
01:44:44,545 --> 01:44:48,082
BUT IN CONTRAST TO THE POLY-A

2043
01:44:48,082 --> 01:44:50,951
HAIRPIN IS MUCH LESS STABLE FOR

2044
01:44:50,951 --> 01:44:52,586
THE 4G RNA.

2045
01:44:52,586 --> 01:44:54,488
THIS IS ABOUT A 90-DEGREE

2046
01:44:54,488 --> 01:44:55,222
DIFFERENCE IN MELTING

2047
01:44:55,222 --> 01:44:55,523
TEMPERATURE.

2048
01:44:55,523 --> 01:44:57,958
IF YOU LOOK AT THE CONTEXT OF

2049
01:44:57,958 --> 01:45:02,029
THE CELL, THE GENOME, THIS IS A

2050
01:45:02,029 --> 01:45:08,903
SITUATION WHERE WE SEE SELECTIVE

2051
01:45:08,903 --> 01:45:09,603
PACKAGING.

2052
01:45:09,603 --> 01:45:12,106
AGAIN IN COLLABORATION WITH

2053
01:45:12,106 --> 01:45:17,545
WEI-SHU AND JONATHAN WE MADE

2054
01:45:17,545 --> 01:45:18,746
DIFFERENT TRANSCRIPTION START

2055
01:45:18,746 --> 01:45:23,584
SITES, MEASURED HAIR SPIN 

2056
01:45:23,584 --> 01:45:23,851
STABILITY.

2057
01:45:23,851 --> 01:45:26,954
WHEN WE CHANGED TO A, COMPARED

2058
01:45:26,954 --> 01:45:32,326
TO NOT NOW 3G BUT GDA, WE SEE

2059
01:45:32,326 --> 01:45:33,961
SOME DIFFERENCES IN MELTING BUT

2060
01:45:33,961 --> 01:45:35,696
LOOK AT THE FIRST DERIVATIVE,

2061
01:45:35,696 --> 01:45:36,797
NOW THERE'S STILL A LITTLE

2062
01:45:36,797 --> 01:45:40,100
DIFFERENCE IN TAR BUT NOW THE

2063
01:45:40,100 --> 01:45:41,402
POLY-A HAIRPIN STABILITY IS

2064
01:45:41,402 --> 01:45:44,138
IDENTICAL, NO LONGER HAS THAT

2065
01:45:44,138 --> 01:45:44,471
DIFFERENCE.

2066
01:45:44,471 --> 01:45:46,974
AND WHEN JONATHAN LOOKED AT

2067
01:45:46,974 --> 01:45:48,609
PACKAGING CELLS, WHAT HE SAW IS

2068
01:45:48,609 --> 01:45:50,844
NO PREFERENTIAL PACKAGING OF ANY

2069
01:45:50,844 --> 01:45:53,013
OF THESE RNAs, ESPECIALLY NOT

2070
01:45:53,013 --> 01:45:55,549
OF THE 1A OR 2A.

2071
01:45:55,549 --> 01:46:00,588
AND SO BUT WE HAVE IDENTIFIED

2072
01:46:00,588 --> 01:46:02,122
ALSO A COMPENSATORY MUTATION,

2073
01:46:02,122 --> 01:46:04,391
BASICALLY IF YOU HAVE THE SAME

2074
01:46:04,391 --> 01:46:08,596
VARIANTS THAT NOW YOU INCLUDE AN

2075
01:46:08,596 --> 01:46:09,563
ADDITIONAL C57 MUTATION, NOW

2076
01:46:09,563 --> 01:46:11,098
WHEN YOU MEASURE MELTING

2077
01:46:11,098 --> 01:46:13,400
TEMPERATURE OF THE TAR POLY A

2078
01:46:13,400 --> 01:46:17,338
YOU SEE WE'VE RESTORED

2079
01:46:17,338 --> 01:46:18,973
DIFFERENCES IN POLY-A HAIRPIN

2080
01:46:18,973 --> 01:46:20,741
STABILITY AND JONATHAN MEASURES

2081
01:46:20,741 --> 01:46:21,175
STABILITY.

2082
01:46:21,175 --> 01:46:22,710
AGAIN, THIS IS CONSISTENT WITH

2083
01:46:22,710 --> 01:46:30,918
THE DIFFERENCES IN POLY-A

2084
01:46:30,918 --> 01:46:33,520
HAIRPIN STABILITY.

2085
01:46:33,520 --> 01:46:35,055
WHAT ABOUT TRANSLATION?

2086
01:46:35,055 --> 01:46:38,125
WE UNDERSTAND PACKAGING WELL,

2087
01:46:38,125 --> 01:46:39,893
WHAT ABOUT TRANSLATION?

2088
01:46:39,893 --> 01:46:41,729
WE PERFORMED IN VITRO AND

2089
01:46:41,729 --> 01:46:42,496
CELL-BASED STUDIES TO LOOK AT

2090
01:46:42,496 --> 01:46:43,264
THIS.

2091
01:46:43,264 --> 01:46:45,666
FOR THE IN VITRO STUDIES WE MADE

2092
01:46:45,666 --> 01:46:48,502
A REPORTER CONSTRUCT, WHICH HAD

2093
01:46:48,502 --> 01:46:52,673
THE FIVE PRIMES UNTRANSLATED

2094
01:46:52,673 --> 01:46:59,146
REGION, A LITTLE BIT OF GAG, AND

2095
01:46:59,146 --> 01:46:59,713
DOWNSTREAM LUCIFERASE GENE,

2096
01:46:59,713 --> 01:47:02,182
TRANSCRIBED IN THE PRESENCE OF

2097
01:47:02,182 --> 01:47:04,385
AUTHENTIC CAP ANALOG, GET BE CAP

2098
01:47:04,385 --> 01:47:04,818
RNA.

2099
01:47:04,818 --> 01:47:06,353
WE TAKE THESE RNAs AND PUT

2100
01:47:06,353 --> 01:47:12,426
THEM INTO IN VITRO TRANSLATION,

2101
01:47:12,426 --> 01:47:14,495
AND LOOK AT THE READOUT.

2102
01:47:14,495 --> 01:47:20,267
BASED ON THE READOUT WE CAN SEE

2103
01:47:20,267 --> 01:47:22,469
IN A CAP-DEPENDENT MANNER, WE

2104
01:47:22,469 --> 01:47:24,838
SEE GREATER TRANSLATION WITH THE

2105
01:47:24,838 --> 01:47:28,609
CAP 3G CONSTRUCT OVER THE CAP 1G

2106
01:47:28,609 --> 01:47:29,043
CONSTRUCT.

2107
01:47:29,043 --> 01:47:31,011
AND SO THIS IS -- THIS WAS

2108
01:47:31,011 --> 01:47:33,747
EXCITING, AND WE WANTED TO MAKE

2109
01:47:33,747 --> 01:47:36,150
SURE -- NOW WE WANT TO TEST OUR

2110
01:47:36,150 --> 01:47:37,484
MUTANTS IN THE SAME SYSTEM.

2111
01:47:37,484 --> 01:47:40,421
HERE AGAIN IS WHAT I JUST SHOWED

2112
01:47:40,421 --> 01:47:42,623
YOU, DIFFERENCE IN READOUT

2113
01:47:42,623 --> 01:47:44,558
BETWEEN THE 1G AND 3G WILD TYPE.

2114
01:47:44,558 --> 01:47:47,928
AND AGAIN YOU CAN SEE THE

2115
01:47:47,928 --> 01:47:49,730
DIFFERENCE IN STRUCTURES ON THE

2116
01:47:49,730 --> 01:47:49,997
GEL.

2117
01:47:49,997 --> 01:47:52,800
IF WE LOOK AT MELTED DIS

2118
01:47:52,800 --> 01:47:54,668
VARIANT, THIS DOES NOT DIMERIZE

2119
01:47:54,668 --> 01:47:56,970
AS EFFICIENTLY BUT HOPEFULLY

2120
01:47:56,970 --> 01:47:58,072
YOU'LL NOTICE CONFIRMATIONAL

2121
01:47:58,072 --> 01:48:03,410
DIFFERENCES IN THE MONOMERS AND

2122
01:48:03,410 --> 01:48:07,381
WITH STILL SEE WE RETRAIN

2123
01:48:07,381 --> 01:48:09,049
PREFERENTIAL TRANSLATION OF 3G

2124
01:48:09,049 --> 01:48:09,583
RNA.

2125
01:48:09,583 --> 01:48:11,985
WE LOOK AT THE MUTANTS I TOLD

2126
01:48:11,985 --> 01:48:15,322
YOU ABOUT EARLIER, SO WHAT WE

2127
01:48:15,322 --> 01:48:18,926
CALL THE GU OR ADD-GG, NOW

2128
01:48:18,926 --> 01:48:19,493
ELIMINATE THE STRUCTURAL

2129
01:48:19,493 --> 01:48:22,096
DIFFERENCES AS YOU CAN SEE ON

2130
01:48:22,096 --> 01:48:25,933
THE NATIVE GEL BETWEEN 1G AND 3G

2131
01:48:25,933 --> 01:48:26,934
REELIMINATE TRANSLATIONAL

2132
01:48:26,934 --> 01:48:27,901
DIFFERENCES, NOW TRANSLATED

2133
01:48:27,901 --> 01:48:30,738
EQUALLY EFFICIENTLY IN THIS IN

2134
01:48:30,738 --> 01:48:33,507
VITRO ASSAY. 

2135
01:48:33,507 --> 01:48:35,209
WHAT ABOUT IN CELLS?

2136
01:48:35,209 --> 01:48:39,279
I WANT TO REMIND EVERYBODY

2137
01:48:39,279 --> 01:48:40,981
TRANSLATION HAPPENS, YOU'LL BE

2138
01:48:40,981 --> 01:48:47,254
SEEING SOME LABELS ON THE GEL,

2139
01:48:47,254 --> 01:48:52,526
SMALL RIBOSOME SCANS, AUG START

2140
01:48:52,526 --> 01:48:56,330
CODON, LARGE SUBUNIT JOINS, YOU

2141
01:48:56,330 --> 01:48:57,765
GET MONOSOMES, ELONGATION

2142
01:48:57,765 --> 01:48:58,031
CONFIDENT.

2143
01:48:58,031 --> 01:49:03,237
YOU WILL START BUILDING UP AS

2144
01:49:03,237 --> 01:49:05,639
TRANSLATION OCCURS LIGHT

2145
01:49:05,639 --> 01:49:06,740
POLYSOMES, TRANSLATING mRNAs

2146
01:49:06,740 --> 01:49:10,244
WILL HAVE MEDIUM AND HEAVY

2147
01:49:10,244 --> 01:49:15,149
POLYSOMES BUILDING ON THE

2148
01:49:15,149 --> 01:49:15,816
mRNA.

2149
01:49:15,816 --> 01:49:21,622
AND SO WEI SUE'S LAB TOOK CELLS,

2150
01:49:21,622 --> 01:49:23,257
TREATED WITH CYCLOHEXIMIDE,

2151
01:49:23,257 --> 01:49:26,093
COLLECTED THE CELL LYSATE,

2152
01:49:26,093 --> 01:49:29,963
LOADED ON SUCROSE DENSITY GRADE

2153
01:49:29,963 --> 01:49:30,197
WENT.

2154
01:49:30,197 --> 01:49:39,139
FRACTIONATED THEM SO WHAT YOU

2155
01:49:39,139 --> 01:49:42,509
SEE HERE, ANALYZE THE RNA BY

2156
01:49:42,509 --> 01:49:46,046
MASS GENERATION FIVE PRIME RACE.

2157
01:49:46,046 --> 01:49:49,650
RESULTS ARE THAT IF YOU LOOK AT

2158
01:49:49,650 --> 01:49:52,052
WHAT'S IN THE TOTAL RNA AS I

2159
01:49:52,052 --> 01:49:52,953
SHOWED BEFORE IT'S MOSTLY 3G

2160
01:49:52,953 --> 01:49:55,189
WITH A LITTLE BIT OF 1G.

2161
01:49:55,189 --> 01:49:57,558
IF YOU LOOK AT THE FREE RNA

2162
01:49:57,558 --> 01:50:00,461
THAT'S NOT BEEN LOADED ON

2163
01:50:00,461 --> 01:50:01,995
POLYSOMES, YOU CAN SEE THAT THAT

2164
01:50:01,995 --> 01:50:03,197
PROFILE DOESN'T CHANGE SO THERE

2165
01:50:03,197 --> 01:50:05,165
DOESN'T SEEM TO BE PREFERENTIAL

2166
01:50:05,165 --> 01:50:07,134
LOADING OF THE 1G OR 3G.

2167
01:50:07,134 --> 01:50:10,637
AND THEN IF YOU LOOK AT THE VERY

2168
01:50:10,637 --> 01:50:13,040
FRACTION, WHAT YOU CAN SEE IS

2169
01:50:13,040 --> 01:50:17,110
THAT THE 1G RNA IS PRESENT IN

2170
01:50:17,110 --> 01:50:20,848
THE 4DF, 6 DF, EVEN THE LIGHT

2171
01:50:20,848 --> 01:50:21,281
POLYSOMES.

2172
01:50:21,281 --> 01:50:26,019
ONCE YOU GOAT MEDIUM AND HEAVY

2173
01:50:26,019 --> 01:50:28,622
IT'S SIGNIFICANTLY LESS

2174
01:50:28,622 --> 01:50:28,956
DOMINANT.

2175
01:50:28,956 --> 01:50:32,893
SO THIS SUGGESTS THE VERY

2176
01:50:32,893 --> 01:50:37,831
ACTIVELY TRANSLATING GENES ARE

2177
01:50:37,831 --> 01:50:40,634
NOT THE 1G RNA.

2178
01:50:40,634 --> 01:50:44,605
WE WANTED -- HERE IS WHAT I

2179
01:50:44,605 --> 01:50:46,807
SHOWED YOU AND QUANTIFICATION,

2180
01:50:46,807 --> 01:50:48,775
YOU CAN SEE PRESENT TO THE LIGHT

2181
01:50:48,775 --> 01:50:49,743
FRACTIONS BUT THEN NOT

2182
01:50:49,743 --> 01:50:52,379
REPRESENTED IN THE MEDIUM AND

2183
01:50:52,379 --> 01:50:53,347
HEAVY FRACTIONS.

2184
01:50:53,347 --> 01:50:57,317
WE WANT TO LOOK AT MUTANTS.

2185
01:50:57,317 --> 01:51:05,425
WHAT WE SAW WITH G MUTANT AND

2186
01:51:05,425 --> 01:51:09,363
ADD-GG, ALL LOADED EQUALLY WELL.

2187
01:51:09,363 --> 01:51:12,399
CHANGING THE STRUCTURE REALLY

2188
01:51:12,399 --> 01:51:14,701
HAS IMPACTS TRANSLATION IN

2189
01:51:14,701 --> 01:51:15,269
CELLS.

2190
01:51:15,269 --> 01:51:17,871
AND SO TO FINISH WITH THINGS

2191
01:51:17,871 --> 01:51:21,608
ONGOING STUDIES WE'RE NOW

2192
01:51:21,608 --> 01:51:25,212
WORKING ON, IF THE 1G -- SORRY,

2193
01:51:25,212 --> 01:51:31,018
THE CAP IS BOUND BY EIF BINDING

2194
01:51:31,018 --> 01:51:34,321
COMPLEX, THAT COMPLEX CONTAINS

2195
01:51:34,321 --> 01:51:35,822
EIF 4E, IN ORDER TO LOCATE THE

2196
01:51:35,822 --> 01:51:38,592
CAP IT'S BEEN SHOWN IN RECENT

2197
01:51:38,592 --> 01:51:42,863
YEARS IT REALLY NEEDS 4G

2198
01:51:42,863 --> 01:51:46,900
SCAFFOLD PROTEIN, AND THE

2199
01:51:46,900 --> 01:51:50,204
HELICASE TO BIND FOR EFFICIENT

2200
01:51:50,204 --> 01:51:52,272
LOADING OF RIBOSOMES, THE FIRST

2201
01:51:52,272 --> 01:51:53,273
STEP.

2202
01:51:53,273 --> 01:51:57,210
LOOK AT CAP BINDING OF 1G AND 3G

2203
01:51:57,210 --> 01:52:03,116
WITH THE COMPLEX, WHICH THIS IS

2204
01:52:03,116 --> 01:52:05,285
WORK DONE IN COLLABORATION,

2205
01:52:05,285 --> 01:52:07,821
PURIFYING 4G FOR US, IN INSECT

2206
01:52:07,821 --> 01:52:10,123
CELLS, YOU SEE SIMILAR BINDING

2207
01:52:10,123 --> 01:52:14,294
BETWEEN THE CAP 1G AND CAP 3G

2208
01:52:14,294 --> 01:52:17,464
RNA CONSISTENT WITH RIBOSOME

2209
01:52:17,464 --> 01:52:18,665
PROFILES, NO PREFERENTIAL

2210
01:52:18,665 --> 01:52:21,068
LOADING OF RIBOSOMES AND NO

2211
01:52:21,068 --> 01:52:23,003
PREFERENTIAL BINDING OF THE

2212
01:52:23,003 --> 01:52:24,571
COMPLEX EITHER.

2213
01:52:24,571 --> 01:52:27,140
AND SO IN COLLABORATION WE'RE

2214
01:52:27,140 --> 01:52:29,843
STARTING  SINGLE MOLECULE

2215
01:52:29,843 --> 01:52:32,312
EXPERIMENTSES TO LOOK AT

2216
01:52:32,312 --> 01:52:34,348
SCANNING OF RIBOSOMES OF THE ONE

2217
01:52:34,348 --> 01:52:36,650
RESULT HERE AGAIN HE'S LOOKING

2218
01:52:36,650 --> 01:52:39,920
AT THE FRET SYSTEM SET UP SO YOU

2219
01:52:39,920 --> 01:52:44,458
HAVE A CAP AND THE ACCEPTOR IS

2220
01:52:44,458 --> 01:52:48,562
DOWNSTREAM OF THE 5 PRIME ATR

2221
01:52:48,562 --> 01:52:52,199
WITH OLIGO, IN THE FOLDED

2222
01:52:52,199 --> 01:52:54,267
CONFIRMATION YOU'D EXPECT A HIGH

2223
01:52:54,267 --> 01:52:57,204
FRET BUT ONCE THIS COMPLEX BINDS

2224
01:52:57,204 --> 01:52:59,406
AND FACILITATES UNWINDING OF THE

2225
01:52:59,406 --> 01:53:01,708
RNA YOU WOULD EXPECT LOW FRETS.

2226
01:53:01,708 --> 01:53:04,878
WE CAN COMPARE THIS FRET

2227
01:53:04,878 --> 01:53:09,916
EFFICIENCY UPON THIS CAP BINDING

2228
01:53:09,916 --> 01:53:16,023
COMPLEX BY NAME, TO ABATE A

2229
01:53:16,023 --> 01:53:17,124
GLOBIN UNSTRUCTURED RNA.

2230
01:53:17,124 --> 01:53:21,828
WE START WITH A HIGH FRET, AS

2231
01:53:21,828 --> 01:53:26,533
YOU ADD THE EIF 4 COMPLEX YOU

2232
01:53:26,533 --> 01:53:27,567
SAY TRANSITION TO COMPLEX IN THE

2233
01:53:27,567 --> 01:53:29,870
LOW FRET STATE.

2234
01:53:29,870 --> 01:53:32,472
HOWEVER WITH 1G AND 3G RNA

2235
01:53:32,472 --> 01:53:34,741
HIGHLY STRUCTURED YOU SEE MORE

2236
01:53:34,741 --> 01:53:37,377
HETEROGENEITY BUT ALSO MUCH

2237
01:53:37,377 --> 01:53:38,945
SLOWER UNWINDING OF THESE RNAs

2238
01:53:38,945 --> 01:53:41,114
AS EXPECTED.

2239
01:53:41,114 --> 01:53:42,082
AGAIN, WE SEE VARIABLES,

2240
01:53:42,082 --> 01:53:43,517
DIFFERENCE BETWEEN ABILITY OF

2241
01:53:43,517 --> 01:53:46,586
THIS COMPLEX TO UNWIND THE 1G

2242
01:53:46,586 --> 01:53:47,421
AND 3G.

2243
01:53:47,421 --> 01:53:49,756
THEY BOTH TAKE 20 OR 30 SECONDS

2244
01:53:49,756 --> 01:53:50,857
TO BE UNWOUND.

2245
01:53:50,857 --> 01:53:52,192
WE'RE INTERESTED IN LOOKING AT

2246
01:53:52,192 --> 01:53:52,626
SCANNING.

2247
01:53:52,626 --> 01:53:55,796
I DON'T HAVE DATA BUT HYPOTHESIS

2248
01:53:55,796 --> 01:53:59,299
BASED ON THE DATA SO FAR IS THAT

2249
01:53:59,299 --> 01:54:05,138
BOTH RNAs CAN LOAD THE CAP

2250
01:54:05,138 --> 01:54:15,048
BINDING COMPLEX AND 4 DIS

2251
01:54:15,048 --> 01:54:15,449
RIBOSOMES.

2252
01:54:15,449 --> 01:54:19,453
RESULTING IN EFFICIENT, YOU

2253
01:54:19,453 --> 01:54:21,188
KNOW, AN EFFICIENT TRANSLATION,

2254
01:54:21,188 --> 01:54:24,357
THOUGH TRANSLATION DOES OCCUR.

2255
01:54:24,357 --> 01:54:28,829
YOU HAVE MORE ON THE LEFT

2256
01:54:28,829 --> 01:54:29,496
STRUCTURED 3G RNA.

2257
01:54:29,496 --> 01:54:31,131
AND SO THAT'S ALL THE SCIENCE I

2258
01:54:31,131 --> 01:54:33,467
WANTED TO TELL YOU ABOUT.

2259
01:54:33,467 --> 01:54:35,769
AGAIN, I MENTIONED A LOT OF

2260
01:54:35,769 --> 01:54:36,837
THESE PEOPLE, SHOWED PICTURES ON

2261
01:54:36,837 --> 01:54:38,472
THE FIRST SLIDE.

2262
01:54:38,472 --> 01:54:40,340
I WON'T BELABOR THIS TO GET TO

2263
01:54:40,340 --> 01:54:41,975
THE IMPORTANT PART.

2264
01:54:41,975 --> 01:54:42,676
THAT'S ALAN.

2265
01:54:42,676 --> 01:54:44,845
A FEW THINGS I'VE LEARNED FROM

2266
01:54:44,845 --> 01:54:46,480
ALAN OVER THE YEARS, NOT A TOP

2267
01:54:46,480 --> 01:54:49,783
TEN BUT A TOP FIVE LIST.

2268
01:54:49,783 --> 01:54:52,519
TOP FIVE, PUT YOUR BOOK DOWN IN

2269
01:54:52,519 --> 01:54:53,386
GRACE AUDITORIUM BEFORE

2270
01:54:53,386 --> 01:54:53,653
BREAKFAST.

2271
01:54:53,653 --> 01:54:55,021
THIS WAS LITERALLY THE FIRST

2272
01:54:55,021 --> 01:54:57,324
THING I LEARNED FROM ALAN.

2273
01:54:57,324 --> 01:55:00,193
AND THE BEST PLACE IS RIGHT

2274
01:55:00,193 --> 01:55:00,427
THERE.

2275
01:55:00,427 --> 01:55:01,495
AND ALAN DOESN'T SIT IN THE

2276
01:55:01,495 --> 01:55:02,162
FRONT ROW.

2277
01:55:02,162 --> 01:55:03,263
HE SITS ABOUT FOUR BACK.

2278
01:55:03,263 --> 01:55:06,199
IF YOU LOOK AT THAT PICTURE THAT

2279
01:55:06,199 --> 01:55:07,400
SOMEONE SHOWED ALREADY THAT'S

2280
01:55:07,400 --> 01:55:09,703
EXACTLY WHERE I WAS SITTING.

2281
01:55:09,703 --> 01:55:11,671
I ALWAYS LIKED SITTING CLOSE TO

2282
01:55:11,671 --> 01:55:14,641
ALAN ESPECIALLY WHEN I FIRST

2283
01:55:14,641 --> 01:55:24,084
STARTED, I'M NOT TRAINED AS A

2284
01:55:24,084 --> 01:55:24,518
VIROLOGISTS.

2285
01:55:24,518 --> 01:55:28,221
I WOULD SAY DO YOU BELIEVE THAT?

2286
01:55:28,221 --> 01:55:31,291
AND WE WOULD HAVE QUIET

2287
01:55:31,291 --> 01:55:31,958
CONVERSATIONS.

2288
01:55:31,958 --> 01:55:33,360
NEXT IS ASK SIMPLE QUESTIONS.

2289
01:55:33,360 --> 01:55:34,394
IMPORTANT THOUGH.

2290
01:55:34,394 --> 01:55:35,796
THESE PAPERS WERE HIGHLIGHTED

2291
01:55:35,796 --> 01:55:39,065
HERE TODAY. 

2292
01:55:39,065 --> 01:55:42,469
WE ALL ADMIRE THE PAPERS. 

2293
01:55:42,469 --> 01:55:46,139
THE QUESTION IS WHAT DO

2294
01:55:46,139 --> 01:55:47,140
RETROVIRAL PACKAGES?

2295
01:55:47,140 --> 01:55:51,278
DELPHINE SHOWED RIBOSOMES,

2296
01:55:51,278 --> 01:55:52,279
SIMPLE QUESTION, AMAZING ANSWER.

2297
01:55:52,279 --> 01:55:57,851
WHAT IS THE KEY COMPONENT OF

2298
01:55:57,851 --> 01:56:01,388
RETROVIRAL ASSEMBLY IN VITRO?

2299
01:56:01,388 --> 01:56:03,990
ERIC OR SOMEONE MENTIONED

2300
01:56:03,990 --> 01:56:04,925
iNOSITOL PHOSPHATES, NOW WE

2301
01:56:04,925 --> 01:56:07,828
KNOW HOW IMPORTANT THEY ARE FOR

2302
01:56:07,828 --> 01:56:11,431
ASSEMBLY IN CELLS.

2303
01:56:11,431 --> 01:56:12,632
BE SKEPTICAL.

2304
01:56:12,632 --> 01:56:13,400
THERE'S A DIFFERENCE BETWEEN

2305
01:56:13,400 --> 01:56:15,735
WHAT YOU KNOW AND WHAT YOU THINK

2306
01:56:15,735 --> 01:56:16,903
YOU KNOW.

2307
01:56:16,903 --> 01:56:19,406
AND RELATED TO THAT, DON'T

2308
01:56:19,406 --> 01:56:19,873
FORGET HISTORY.

2309
01:56:19,873 --> 01:56:23,610
IF YOU WEREN'T THERE FOR ALAN'S

2310
01:56:23,610 --> 01:56:25,478
COLD SPRING HARBOR KEYNOTE TALK

2311
01:56:25,478 --> 01:56:27,981
IN 2016, FIND IT ON YouTube

2312
01:56:27,981 --> 01:56:30,383
AND LISTEN TO THAT, I LEARNED A

2313
01:56:30,383 --> 01:56:31,151
LOT FROM IT.

2314
01:56:31,151 --> 01:56:32,552
AND THEN THIS ONE.

2315
01:56:32,552 --> 01:56:34,554
IT'S OKAY TO JUST LOOK OUT THE

2316
01:56:34,554 --> 01:56:34,821
WINDOW.

2317
01:56:34,821 --> 01:56:36,623
THIS IS RELATED TO ONE TIME ALAN

2318
01:56:36,623 --> 01:56:39,125
AND I HAD BEEN AT MANY

2319
01:56:39,125 --> 01:56:40,760
CONFERENCES, MANY TIMES IN

2320
01:56:40,760 --> 01:56:43,930
EUROPE OR WHATEVER.

2321
01:56:43,930 --> 01:56:48,101
YEAH, I DON'T MIND LONG FLIGHTS,

2322
01:56:48,101 --> 01:56:50,203
GET WORK DONE, WRITE PAPERS.

2323
01:56:50,203 --> 01:56:52,572
HE SAID, YOU KNOW, I JUST LIKE

2324
01:56:52,572 --> 01:56:54,507
TO LOOK OUT THE WINDOW.

2325
01:56:54,507 --> 01:56:56,009
THIS IS A PICTURE I TOOK LAST

2326
01:56:56,009 --> 01:56:58,745
YEAR ON THE WAY TO COLD SPRING

2327
01:56:58,745 --> 01:57:02,582
HARBOR, A BEAUTIFUL DAY, I

2328
01:57:02,582 --> 01:57:05,318
SHOULD CAPTURE THAT.

2329
01:57:05,318 --> 01:57:07,921
OVER THE YEARS WE HAD PUBLISHED

2330
01:57:07,921 --> 01:57:09,923
EIGHT MANUSCRIPTS AND ARTICLES

2331
01:57:09,923 --> 01:57:12,626
OF VARIOUS KINDS TOGETHER.

2332
01:57:12,626 --> 01:57:14,160
WE SHARED COMMON INTERESTS, WHEN

2333
01:57:14,160 --> 01:57:16,897
WE HAD AN OPPORTUNITY WE TRIED

2334
01:57:16,897 --> 01:57:17,898
TO COLLABORATE TOGETHER, AND

2335
01:57:17,898 --> 01:57:19,399
IT'S BEEN A LOT OF FUN.

2336
01:57:19,399 --> 01:57:21,401
WE HAVE A LOT OF FOND MEMORIES

2337
01:57:21,401 --> 01:57:25,538
LIKE I SAID ALL OVER EUROPE AND

2338
01:57:25,538 --> 01:57:26,106
EVERYWHERE.

2339
01:57:26,106 --> 01:57:28,275
BUT THIS WAS THE BOSTON DUCK

2340
01:57:28,275 --> 01:57:32,245
BOAT TOUR IN 2019 AT THE

2341
01:57:32,245 --> 01:57:33,747
RETROVIRAL ASSEMBLY MEETING.

2342
01:57:33,747 --> 01:57:36,316
AND WE WERE SITTING RIGHT BEHIND

2343
01:57:36,316 --> 01:57:38,118
I THINK KATHY AND BEN, THAT WAS

2344
01:57:38,118 --> 01:57:38,451
FUN.

2345
01:57:38,451 --> 01:57:46,226
LOTS OF FUN DINNERS TOGETHER.

2346
01:57:46,226 --> 01:57:48,862
THIS WAS AFTER THE MEETING WITH

2347
01:57:48,862 --> 01:57:52,265
OUR MUTUAL AND DEAR FRIEND

2348
01:57:52,265 --> 01:57:54,167
JUDITH.

2349
01:57:54,167 --> 01:57:56,536
WE INTERACTED A LOT, COLD SPRING

2350
01:57:56,536 --> 01:57:58,471
HARBOR, HERE AT MY SABBATICAL

2351
01:57:58,471 --> 01:57:58,939
AND MEETINGS.

2352
01:57:58,939 --> 01:58:00,807
THIS IS ONE MEETING I DIDN'T GO

2353
01:58:00,807 --> 01:58:02,909
TO BECAUSE I CAN'T FIND MYSELF

2354
01:58:02,909 --> 01:58:05,445
BUT I FOUND ALAN, YOU'LL NOTICE

2355
01:58:05,445 --> 01:58:07,614
IN THE CENTER.

2356
01:58:07,614 --> 01:58:09,683
SO YOU KNOW HOW THEY SAY LET'S

2357
01:58:09,683 --> 01:58:12,886
GET UP, GET A PICTURE, WE

2358
01:58:12,886 --> 01:58:13,353
ASSEMBLE.

2359
01:58:13,353 --> 01:58:16,256
EVERY PHOTO I FOUND, YOU SEE HIM

2360
01:58:16,256 --> 01:58:18,024
ON THE BOTTOM HERE?

2361
01:58:18,024 --> 01:58:20,093
IN THE CENTER AGAIN.

2362
01:58:20,093 --> 01:58:21,728
THIS IS MINNEAPOLIS MEETING.

2363
01:58:21,728 --> 01:58:24,030
HERE'S A COUPLE MORE.

2364
01:58:24,030 --> 01:58:26,199
THIS ONE HE'S MAYBE OFF CENTER

2365
01:58:26,199 --> 01:58:30,470
BUT PRETTY CLOSE, A STRANGE

2366
01:58:30,470 --> 01:58:31,338
L-SHAPED ROOM.

2367
01:58:31,338 --> 01:58:34,074
LAST ONE, BOSTON, THERE HE IS.

2368
01:58:34,074 --> 01:58:37,911
TO THE PERSON, I FOUND IT.

2369
01:58:37,911 --> 01:58:39,779
THIS REALLY IS BECAUSE HE'S A

2370
01:58:39,779 --> 01:58:41,648
CENTRAL IF NOT THE CENTRAL

2371
01:58:41,648 --> 01:58:45,251
PLAYER IN THIS FIELD OF

2372
01:58:45,251 --> 01:58:47,988
RETROVIRAL ASSEMBLY SO IT'S

2373
01:58:47,988 --> 01:58:48,521
APPROPRIATE.

2374
01:58:48,521 --> 01:58:51,591
I COULDN'T FIND THE LAST PHOTO

2375
01:58:51,591 --> 01:58:52,993
FROM THE MEETING, MANY OF YOU

2376
01:58:52,993 --> 01:58:55,628
WERE THERE IN SALT LAKE CITY.

2377
01:58:55,628 --> 01:58:58,164
I E-MAILED, DO YOU HAVE THE

2378
01:58:58,164 --> 01:58:58,832
GROUP PHOTO?

2379
01:58:58,832 --> 01:59:00,233
WE FORGOT TO TAKE ONE.

2380
01:59:00,233 --> 01:59:03,636
THEN I REMEMBERED FOR THOSE THAT

2381
01:59:03,636 --> 01:59:07,907
WERE THERE, THIS PHYSICIST HAS

2382
01:59:07,907 --> 01:59:10,443
CONNECTIONS WITH ASTRONOMERS.

2383
01:59:10,443 --> 01:59:11,745
AFTER THE POSTER SESSION,

2384
01:59:11,745 --> 01:59:16,783
ASTRONOMY SET UP TELESCOPES IN

2385
01:59:16,783 --> 01:59:22,022
PITCH DARK UTAH AND LOOKED UP AT

2386
01:59:22,022 --> 01:59:22,555
CELESTIAL BODIES.

2387
01:59:22,555 --> 01:59:23,656
THIS IS APPROXIMATELY WHAT WE

2388
01:59:23,656 --> 01:59:23,823
SAW.

2389
01:59:23,823 --> 01:59:27,994
IN THE CENTER WE ALSO HAVE ALAN.

2390
01:59:27,994 --> 01:59:30,563
[LAUGHTER] 

2391
01:59:30,563 --> 01:59:36,236
SO,  ALAN, THANK YOU FOR BEING

2392
01:59:36,236 --> 01:59:38,438
THE CENTER OF OUR UNIVERSE AND

2393
01:59:38,438 --> 01:59:40,673
THE RETROVIRUS FIELD, AND

2394
01:59:40,673 --> 01:59:50,984
CONGRATULATIONS. 

2395
01:59:56,823 --> 01:59:59,025
>> SO YOUR COMMENTS ABOUT
HIV,

2396
01:59:59,025 --> 02:00:01,761
HAVE YOU EVER BEEN ABLE TO

2397
02:00:01,761 --> 02:00:04,397
MEASURE EFFECTS OF, SAY, THESE

2398
02:00:04,397 --> 02:00:09,636
MUTATIONS ON VIRUS REPLICATION? 

2399
02:00:09,636 --> 02:00:09,903
>> YES.

2400
02:00:09,903 --> 02:00:11,604
SO, WEI-SUH CAN ANSWER BETTER

2401
02:00:11,604 --> 02:00:17,844
THAN I CAN BUT BASICALLY IF YOU

2402
02:00:17,844 --> 02:00:22,315
HAVE -- IF YOU ALTER DISTANCE,

2403
02:00:22,315 --> 02:00:25,618
HAVE BEEN ABLE TO MAKE A 1G ONLY

2404
02:00:25,618 --> 02:00:29,856
VIRUS AND 3G ONLY VIRUS, ONLY

2405
02:00:29,856 --> 02:00:31,057
EXPRESS AND ARE REPLICATION

2406
02:00:31,057 --> 02:00:31,324
COMPETENT.

2407
02:00:31,324 --> 02:00:33,326
IF YOU COMPETE THEM WITH THE

2408
02:00:33,326 --> 02:00:34,461
WILD TYPE, THE WILD TYPE ALWAYS

2409
02:00:34,461 --> 02:00:35,895
WINS OUT.

2410
02:00:35,895 --> 02:00:39,833
THERE'S A FITNESS DEFECT.

2411
02:00:39,833 --> 02:00:45,405
AND SO, YEAH, ANYTHING ELSE,

2412
02:00:45,405 --> 02:00:46,272
WEI-SUH?

2413
02:00:46,272 --> 02:00:48,241
OKAY.

2414
02:00:48,241 --> 02:00:51,077
THE VIRUS HAS OPTIMIZED -- TAKEN

2415
02:00:51,077 --> 02:00:53,713
ADVANTAGE OF THE SLOPPINESS OF

2416
02:00:53,713 --> 02:00:55,148
THE HOST RNA POLYMERASE.

2417
02:00:55,148 --> 02:00:56,483
I GUESS THAT'S STILL A

2418
02:00:56,483 --> 02:01:01,054
POSSIBILITY BUT WE DON'T KNOW

2419
02:01:01,054 --> 02:01:02,155
THAT.

2420
02:01:02,155 --> 02:01:06,693
ADAM?

2421
02:01:06,693 --> 02:01:11,798
>> REAL BEAUTIFUL WORK,

2422
02:01:11,798 --> 02:01:18,037
REMARKABLE PHOTOGRAPH. 

2423
02:01:18,037 --> 02:01:21,641
IS THERE A WAY YOU CAN

2424
02:01:21,641 --> 02:01:22,108
MANIPULATE IT?

2425
02:01:22,108 --> 02:01:24,611
>> I BET YOU COULD.

2426
02:01:24,611 --> 02:01:27,914
WE HAVEN'T TRIED THAT. 

2427
02:01:27,914 --> 02:01:30,884
DIMERIZATION, THE VARIANT THAT

2428
02:01:30,884 --> 02:01:32,785
DOESN'T -- IT STILL FORMS A

2429
02:01:32,785 --> 02:01:33,686
DIMER, STILL PACKAGES

2430
02:01:33,686 --> 02:01:35,955
SELECTIVELY AND EVERYTHING BUT

2431
02:01:35,955 --> 02:01:39,459
WE DO THINK DIMERIZATION IS

2432
02:01:39,459 --> 02:01:40,326
IMPORTANT.

2433
02:01:40,326 --> 02:01:45,498
WE'VE NEVER TRIED THAT

2434
02:01:45,498 --> 02:01:47,567
SPECIFICALLY.

2435
02:01:47,567 --> 02:01:48,334
>> (INAUDIBLE).

2436
02:01:48,334 --> 02:01:50,970
>> UH-HUH, YEAH.

2437
02:01:50,970 --> 02:01:53,706
>> WHAT WAS THE RATIO OF

2438
02:01:53,706 --> 02:01:56,943
INVESTIGATORS TO AMINO ACIDS AT

2439
02:01:56,943 --> 02:02:00,947
THE MEETING?

2440
02:02:00,947 --> 02:02:03,016
[LAUGHTER]

2441
02:02:03,016 --> 02:02:03,650
>> ABOUT TWO INVESTIGATORS
FOR

2442
02:02:03,650 --> 02:02:04,751
EVERY AMINO ACID.

2443
02:02:04,751 --> 02:02:09,022
I SHOULD SAY MAYBE ERIC WOULD

2444
02:02:09,022 --> 02:02:11,090
HAVE A SLIDE, THE NEXT ONE IS

2445
02:02:11,090 --> 02:02:15,061
GOING TO BE IN PRAGUE, END OF I

2446
02:02:15,061 --> 02:02:16,896
BELIEVE -- IS IT MIDDLE OR END

2447
02:02:16,896 --> 02:02:17,430
OF SEPTEMBER?

2448
02:02:17,430 --> 02:02:19,432
>> THIRD WEEK.

2449
02:02:19,432 --> 02:02:20,533
>> THIRD WEEK IN
SEPTEMBER,

2450
02:02:20,533 --> 02:02:20,833
2025.

2451
02:02:20,833 --> 02:02:23,903
WE'LL GET AN E-MAIL OUT,

2452
02:02:23,903 --> 02:02:24,771
ORGANIZING IT, HOPEFULLY NEXT

2453
02:02:24,771 --> 02:02:29,809
WEEK FOR THOSE ON THE LIST.

2454
02:02:29,809 --> 02:02:31,144
>> BEAUTIFUL.

2455
02:02:31,144 --> 02:02:36,716
I'M TRYING TO REMEMBER ABOUT

2456
02:02:36,716 --> 02:02:38,384
PACKAGING, MLBs, DO OTHER

2457
02:02:38,384 --> 02:02:39,586
VIRUSES DO SELECTIVE PACKAGE?

2458
02:02:39,586 --> 02:02:43,089
>> THERE'S ALSO GOING TO
BE TWO

2459
02:02:43,089 --> 02:02:44,624
PULLS, AND WITH HIV IT WAS NEVER

2460
02:02:44,624 --> 02:02:47,026
THOUGHT TO BE THE CASE.

2461
02:02:47,026 --> 02:02:49,629
BUT IN RECENT YEARS IT'S

2462
02:02:49,629 --> 02:02:50,930
SHOWN -- THE MECHANISM IS

2463
02:02:50,930 --> 02:02:53,233
DIFFERENT IN MLV.

2464
02:02:53,233 --> 02:02:54,667
WE DON'T THINK IT'S RELATED TO

2465
02:02:54,667 --> 02:02:55,868
THE FIVE PRIME END.

2466
02:02:55,868 --> 02:03:04,177
YES, IT'S BEEN SHOWN IN OTHER

2467
02:03:04,177 --> 02:03:06,879
VIRUSES.

2468
02:03:06,879 --> 02:03:07,480
DELPHINE?

2469
02:03:07,480 --> 02:03:12,285
>> I WAS WONDERING IF YOU
HAVE

2470
02:03:12,285 --> 02:03:18,858
DONE OR INTEND TO DO STRUCTURE

2471
02:03:18,858 --> 02:03:19,959
WITH SENTAG DYNAMIC OF

2472
02:03:19,959 --> 02:03:21,794
TRANSLATION IN THE CELL.

2473
02:03:21,794 --> 02:03:22,695
>> YES.

2474
02:03:22,695 --> 02:03:28,334
>> AND YOU CAN SEE MAYBE
IT'S --

2475
02:03:28,334 --> 02:03:33,573
>> IT'S IN MY PROPOSAL.

2476
02:03:33,573 --> 02:03:34,807
YEAH, NO, THOSE ARE EXACTLY THE

2477
02:03:34,807 --> 02:03:36,342
THINGS WE WANT TO DO.

2478
02:03:36,342 --> 02:03:37,543
WE UNDERSTAND THE BASICS.

2479
02:03:37,543 --> 02:03:40,847
WHAT ARE THE DIFFERENCES IN THE

2480
02:03:40,847 --> 02:03:44,584
DYNAMICS, KINETICS, PROTEIN

2481
02:03:44,584 --> 02:03:45,018
BINDING DIFFERENCES,

2482
02:03:45,018 --> 02:03:45,885
METHYLATION, MODIFICATION, WE

2483
02:03:45,885 --> 02:03:49,055
HAVE THE TOOLS BUT THE 1G,

2484
02:03:49,055 --> 02:03:53,126
3G-ONLY VIRUS, TO LOOK AT THAT.

2485
02:03:53,126 --> 02:03:59,232
>> YOU COULD SEE THE RNA
-- I

2486
02:03:59,232 --> 02:04:01,334
MEAN TRANSLATION, BUT THEN DO

2487
02:04:01,334 --> 02:04:03,303
YOU THINK THIS GAG WOULD BE THE

2488
02:04:03,303 --> 02:04:06,339
CHAPERONE OR ONE OR THE OTHER OF

2489
02:04:06,339 --> 02:04:09,642
THE SPECIES OVER TIME?

2490
02:04:09,642 --> 02:04:10,410
>> A TRANSLATION
CHAPERONE?

2491
02:04:10,410 --> 02:04:12,145
WHAT DO YOU MEAN?

2492
02:04:12,145 --> 02:04:13,780
>> WE KNOW GAG, LIKE I
SAID IN

2493
02:04:13,780 --> 02:04:16,182
VITRO, WE ALSO WANT TO LOOK AT

2494
02:04:16,182 --> 02:04:18,351
ANY IN CELLS BUT DEFINITELY

2495
02:04:18,351 --> 02:04:20,586
ASSEMBLE MORE READILY IN THE

2496
02:04:20,586 --> 02:04:22,555
FIVE PRIME OF THE 1G RNA, I

2497
02:04:22,555 --> 02:04:26,693
IMAGINE THE SAME THING IS TRUE

2498
02:04:26,693 --> 02:04:28,127
IN CELLS.

2499
02:04:28,127 --> 02:04:28,594
WE HAVEN'T LOOKED-IT.

2500
02:04:28,594 --> 02:04:30,797
I DON'T THINK IT'S A SWITCH.

2501
02:04:30,797 --> 02:04:33,299
I THINK IT'S TWO SPECIES, RIGHT?

2502
02:04:33,299 --> 02:04:34,734
ONE OF THEM, EXACTLY, YOU KNOW,

2503
02:04:34,734 --> 02:04:37,270
ONE OF THEM IS BOUND BY GAG

2504
02:04:37,270 --> 02:04:39,405
READILY AND GUIDED TO THE

2505
02:04:39,405 --> 02:04:41,441
MEMBRANE FOR ASSEMBLY.

2506
02:04:41,441 --> 02:04:46,746
THE OTHER ONE -- BOTH LOADED BY

2507
02:04:46,746 --> 02:04:49,148
RIBOSOMES. 

2508
02:04:49,148 --> 02:04:57,023
THE 1G IS NOT AS EFFECTIVE, IT'S

2509
02:04:57,023 --> 02:05:01,627
A DISTRIBUTION OF  ENSEMBLES OF

2510
02:05:01,627 --> 02:05:02,595
RNA STRUCTURES, DISTRIBUTION

2511
02:05:02,595 --> 02:05:08,067
SHIFTED AND FOR THESE DIFFERENT

2512
02:05:08,067 --> 02:05:08,568
FUNCTIONS.

2513
02:05:08,568 --> 02:05:12,238
>> I KNOW THERE'S
DIFFERENCES IN

2514
02:05:12,238 --> 02:05:16,609
CAP BINDING COMPLEXES.

2515
02:05:16,609 --> 02:05:16,943
>> YES.

2516
02:05:16,943 --> 02:05:22,949
>> SO IS ANYTHING KNOWN IN
T

2517
02:05:22,949 --> 02:05:24,317
CELLS SPECIFICALLY ABOUT --

2518
02:05:24,317 --> 02:05:27,553
>> EXACTLY WHAT THOSE ARE
IN T

2519
02:05:27,553 --> 02:05:27,754
CELLS?

2520
02:05:27,754 --> 02:05:29,288
I DON'T KNOW.

2521
02:05:29,288 --> 02:05:31,023
I THOUGHT YOU WERE GOING ASK

2522
02:05:31,023 --> 02:05:34,594
ABOUT THE TRIMETHYL G CAP.

2523
02:05:34,594 --> 02:05:36,662
IT'S BEEN PROPOSED AND SHOWN

2524
02:05:36,662 --> 02:05:38,931
PRETTY CLEARLY TO BE SOMETIMES

2525
02:05:38,931 --> 02:05:42,769
GIVE US TRIMETHYL G CAP AND BIND

2526
02:05:42,769 --> 02:05:44,103
ALTERNATIVE COMPLEX AND WE'RE

2527
02:05:44,103 --> 02:05:46,305
LOOKING AT THAT BUT IN TERMS OF

2528
02:05:46,305 --> 02:05:52,845
IN T CELLS WHAT THE ISOFORMS ARE

2529
02:05:52,845 --> 02:05:54,380
WE HAVEN'T LOOKED.

2530
02:05:54,380 --> 02:06:04,791
I I SHOULD STOP THERE. 

2531
02:06:06,726 --> 02:06:16,702
 

2532
02:06:22,975 --> 02:06:25,111
>> THANK YOU FOR A
BEAUTIFUL

2533
02:06:25,111 --> 02:06:25,678
TALK, KARIN.

2534
02:06:25,678 --> 02:06:28,114
WE'VE HEARD THE FIRST PART OF MY

2535
02:06:28,114 --> 02:06:30,082
INTRODUCTION OF THE NEXT

2536
02:06:30,082 --> 02:06:35,588
SPEAKER, BY MISTAKE, BUT THE

2537
02:06:35,588 --> 02:06:37,190
NEXT SPEAKER IS MAURICIO

2538
02:06:37,190 --> 02:06:39,358
COMAS-GARCIA, WHO WAS ALSO A

2539
02:06:39,358 --> 02:06:44,397
POSTDOC IN MY LAB, ACTUALLY

2540
02:06:44,397 --> 02:06:48,701
PRETTY RECENTLY. 

2541
02:06:48,701 --> 02:06:51,971
HE CAME FROM A LAB REALLY FROM A

2542
02:06:51,971 --> 02:06:54,507
PHYSICS BACKGROUND, FROM A

2543
02:06:54,507 --> 02:06:58,110
COMMUNITY OF PHYSICISTS WHO HAD

2544
02:06:58,110 --> 02:07:01,280
DECIDED TO STUDY VIRUSES.

2545
02:07:01,280 --> 02:07:06,185
CENTERED AROUND UCLA, WHO WAS

2546
02:07:06,185 --> 02:07:07,220
HIS THESIS ADVISER, MAURICIO

2547
02:07:07,220 --> 02:07:10,723
CAME TO MY LAB AND REALLY WORKED

2548
02:07:10,723 --> 02:07:13,893
OUT THE TECHNIQUES TO MEASURE

2549
02:07:13,893 --> 02:07:18,698
BINDING OF GAG TO DIFFERENT

2550
02:07:18,698 --> 02:07:20,433
RNAs, UNDER DIFFERENT

2551
02:07:20,433 --> 02:07:21,400
CONDITIONS.

2552
02:07:21,400 --> 02:07:23,169
I THINK HE DID BEAUTIFUL WORK

2553
02:07:23,169 --> 02:07:28,107
FOLLOWING THE FOOT STEPS OF WHAT

2554
02:07:28,107 --> 02:07:29,842
KARIN HAD ALREADY PUBLISHED,

2555
02:07:29,842 --> 02:07:33,913
EXPANDED THE VIEW WITH

2556
02:07:33,913 --> 02:07:34,780
FASCINATING RESULTS.

2557
02:07:34,780 --> 02:07:36,949
I WON'T TALK ABOUT THAT, I'LL

2558
02:07:36,949 --> 02:07:40,887
JUST SAY HE DID BEAUTIFUL WORK

2559
02:07:40,887 --> 02:07:44,524
IN MY LAB AND WAS DELIGHTED TO

2560
02:07:44,524 --> 02:07:47,593
GET A NICE JOB IN HIS HOME

2561
02:07:47,593 --> 02:07:51,397
COUNTRY, WHICH IS MEXICO, AND SO

2562
02:07:51,397 --> 02:07:59,839
HE'S NOW PROFESSOR IN SON SAN

2563
02:07:59,839 --> 02:08:00,873
LUIS POTOSI CONTINUING TO DO

2564
02:08:00,873 --> 02:08:02,675
GREAT WORK IN A NUMBER OF

2565
02:08:02,675 --> 02:08:03,242
DIRECTIONS.

2566
02:08:03,242 --> 02:08:04,176
SORRY HE WOULDN'T BE HERE IN

2567
02:08:04,176 --> 02:08:07,914
PERSON BUT THERE HE IS.

2568
02:08:07,914 --> 02:08:18,391
GOOD MORNING,  BUENOS DIAS.

2569
02:08:18,658 --> 02:08:23,195
LOOK FORWARD TO YOUR TOOK.

2570
02:08:23,195 --> 02:08:25,498
>> IT'S A SHAME UNITED
AIRLINES

2571
02:08:25,498 --> 02:08:29,201
DIDN'T GET IT TOGETHER.

2572
02:08:29,201 --> 02:08:33,472
AND NOW I GUESS YOU CAN SEE MY

2573
02:08:33,472 --> 02:08:38,945
SCREEN RIGHT NOW.

2574
02:08:38,945 --> 02:08:39,912
OKAY.

2575
02:08:39,912 --> 02:08:42,114
SO, FIRST THIS IS ALAN WHEN HE

2576
02:08:42,114 --> 02:08:51,424
CAME TO VISIT AFTER I MOVED.

2577
02:08:51,424 --> 02:08:52,625
SIX MONTHS.

2578
02:08:52,625 --> 02:08:54,794
I'M DEEPLY SADDENED NOT ABLE TO

2579
02:08:54,794 --> 02:08:56,996
ATTEND TO THE MEETING IN PERSON.

2580
02:08:56,996 --> 02:09:00,466
BUT WE'RE HERE TO CELEBRATE YOU,

2581
02:09:00,466 --> 02:09:02,134
YOUR REMARKABLE SCIENTIFIC

2582
02:09:02,134 --> 02:09:03,002
CAREER, MOST IMPORTANTLY I'M

2583
02:09:03,002 --> 02:09:06,072
HERE TO THANK YOU FOR EVERYTHING

2584
02:09:06,072 --> 02:09:07,506
YOU'VE SAID TO ME.

2585
02:09:07,506 --> 02:09:10,343
I'M SORRY I WON'T BE TALKING

2586
02:09:10,343 --> 02:09:11,310
ABOUT HIV.

2587
02:09:11,310 --> 02:09:13,412
BUT BE SURE EVERYTHING I WORKED

2588
02:09:13,412 --> 02:09:17,016
ON IS A SEQUENCE OF YOUR

2589
02:09:17,016 --> 02:09:18,351
PREVIOUS RESEARCH.

2590
02:09:18,351 --> 02:09:20,620
YOU TAUGHT ME TO BE KIND,

2591
02:09:20,620 --> 02:09:22,388
POLITE, PASSIONATE.

2592
02:09:22,388 --> 02:09:24,790
YOU TAUGHT ME THAT WORDS MATTER,

2593
02:09:24,790 --> 02:09:28,527
ONE SHOULD BE CAREFUL WHEN

2594
02:09:28,527 --> 02:09:30,796
WRITING, HAVING SCIENTIFIC

2595
02:09:30,796 --> 02:09:31,097
DISCUSSIONS.

2596
02:09:31,097 --> 02:09:32,865
BUT HE ALSO TAUGHT ME TO BE

2597
02:09:32,865 --> 02:09:33,766
VERY, VERY SKEPTICAL.

2598
02:09:33,766 --> 02:09:36,402
YOU TAUGHT ME IT'S OKAY TO SAY I

2599
02:09:36,402 --> 02:09:37,370
DON'T KNOW.

2600
02:09:37,370 --> 02:09:39,238
YOU TAUGHT ME THAT ONE OF THE

2601
02:09:39,238 --> 02:09:43,509
GREATEST PLEASURES IN LIFE IS TO

2602
02:09:43,509 --> 02:09:45,478
DEEPLY UNDERSTAND WHAT YOU

2603
02:09:45,478 --> 02:09:46,879
STARTED, WHAT YOU THINK IS

2604
02:09:46,879 --> 02:09:47,179
INTERESTING.

2605
02:09:47,179 --> 02:09:49,849
IF I HAD TO CHOOSE MY CAREER

2606
02:09:49,849 --> 02:09:52,151
PATH AGAIN, FOR SURE I WOULD

2607
02:09:52,151 --> 02:09:54,587
CHOOSE WORKING WITH YOU.

2608
02:09:54,587 --> 02:09:56,122
HOWEVER, I MIGHT HAVE FOUND A

2609
02:09:56,122 --> 02:09:59,725
WAY TO STAY LONGER IN YOUR LAB.

2610
02:09:59,725 --> 02:10:02,128
IN FACT A COUPLE WEEKS AGO, I

2611
02:10:02,128 --> 02:10:04,296
SAW BILL, HE TOLD ME AFTER

2612
02:10:04,296 --> 02:10:06,065
SPENDING A FEW DAYS WORKING

2613
02:10:06,065 --> 02:10:07,833
TOGETHER, I'M GLAD YOU WENT TO

2614
02:10:07,833 --> 02:10:09,769
WORK WITH ALAN.

2615
02:10:09,769 --> 02:10:12,705
NOW YOU ARE REAL VIROLOGIST,

2616
02:10:12,705 --> 02:10:13,839
WHATEVER THAT MEANS, I DON'T

2617
02:10:13,839 --> 02:10:15,274
KNOW WHAT THAT MEANS.

2618
02:10:15,274 --> 02:10:17,109
ALAN, THERE ARE NO WORDS TO

2619
02:10:17,109 --> 02:10:20,613
EXPRESS HOW MUCH I ADMIRE YOU, I

2620
02:10:20,613 --> 02:10:23,649
OWE YOU MORE THAN I COULD EVER

2621
02:10:23,649 --> 02:10:25,751
GIVE YOU BACK.

2622
02:10:25,751 --> 02:10:29,689
ONE OF MY FAVORITE MOVIES, OH

2623
02:10:29,689 --> 02:10:31,757
CAPTAIN, MY CAPTAIN, YOU HAVE A

2624
02:10:31,757 --> 02:10:33,392
FRIEND FOR LIFE.

2625
02:10:33,392 --> 02:10:37,129
SO I WANT TO FIRST -- I WROTE A

2626
02:10:37,129 --> 02:10:43,035
FEW WEEKS AGO, I'M SORRY, I

2627
02:10:43,035 --> 02:10:49,508
WON'T TALK ABOUT HIV, BUT I'LL

2628
02:10:49,508 --> 02:10:51,377
TALK ABOUT ALPHAVIRUSES.

2629
02:10:51,377 --> 02:10:54,413
AND ALPHAVIRUSES ARE VERY, VERY

2630
02:10:54,413 --> 02:10:56,082
INTERESTING BECAUSE THEY ALLOW

2631
02:10:56,082 --> 02:10:57,483
ME TO KEEP STUDYING ASSEMBLY IN

2632
02:10:57,483 --> 02:10:59,351
A VERY DIFFERENT WAY FROM WHAT

2633
02:10:59,351 --> 02:11:03,522
DID I WITH ALAN.

2634
02:11:03,522 --> 02:11:05,391
THESE VIRUSES ARE AMAZING, TWO

2635
02:11:05,391 --> 02:11:08,427
LAYERS, SEPARATED BY A LIPID

2636
02:11:08,427 --> 02:11:09,195
BILAYER.

2637
02:11:09,195 --> 02:11:13,365
YOU CAN SEE HERE THE CORE

2638
02:11:13,365 --> 02:11:15,868
CONTAINS NUCLEIC ACID, 240

2639
02:11:15,868 --> 02:11:18,304
COPIES OF THE CAPSID PROTEIN.

2640
02:11:18,304 --> 02:11:19,872
OUTER LAYER CON TAINTS 80

2641
02:11:19,872 --> 02:11:22,241
TRIMERS OF HETERODIMERS OF THE

2642
02:11:22,241 --> 02:11:22,808
GLYCOPROTEINS.

2643
02:11:22,808 --> 02:11:24,210
TODAY I'M GOING TO TELL YOU

2644
02:11:24,210 --> 02:11:26,112
ABOUT A SMALL PROJECT TO

2645
02:11:26,112 --> 02:11:27,179
UNDERSTAND THE ASSEMBLY OF THE

2646
02:11:27,179 --> 02:11:31,550
CORE BECAUSE WHEN YOU TAKE THE

2647
02:11:31,550 --> 02:11:34,520
CURRENT DATA AND REMOVE DENSITY

2648
02:11:34,520 --> 02:11:37,022
FOR THE PLASMA MEMBRANE, WELL

2649
02:11:37,022 --> 02:11:39,859
VIRAL MEMBRANE AND GLYCOPROTEINS

2650
02:11:39,859 --> 02:11:45,998
YOU SEE THESE WIFFLE BALLS WITH

2651
02:11:45,998 --> 02:11:47,767
LARGE HOLES AND DISTANCES WHICH

2652
02:11:47,767 --> 02:11:48,400
BAFFLES ME.

2653
02:11:48,400 --> 02:11:53,005
TWENTY YEARS AGO OR MORE,

2654
02:11:53,005 --> 02:11:56,742
ANOTHER PERSON I ADMIRE A LOT

2655
02:11:56,742 --> 02:12:00,813
PUBLISHED THIS PAPER, SHE WAS

2656
02:12:00,813 --> 02:12:03,983
TRYING TO RECONSTRUCT AGAIN THE

2657
02:12:03,983 --> 02:12:05,618
CORE OF TWO ALPHAVIRUSES.

2658
02:12:05,618 --> 02:12:07,253
THIS IS AMAZINGLY SHE WAS ABLE

2659
02:12:07,253 --> 02:12:09,789
TO ABLE BECAUSE SHE COULDN'T DO

2660
02:12:09,789 --> 02:12:14,593
IT TO RESOLUTION BETTER THAN 30

2661
02:12:14,593 --> 02:12:19,732
ANGSTROMS, AT THAT TIME IT WAS 8

2662
02:12:19,732 --> 02:12:20,432
ANGSTROMS.

2663
02:12:20,432 --> 02:12:23,803
OFTEN SHE WAS COMPLETELY RIGHT,

2664
02:12:23,803 --> 02:12:25,871
INABILITY TO IMPROVE RESOLUTION

2665
02:12:25,871 --> 02:12:27,506
SUGGESTS CORE-LIKER PARTICLES

2666
02:12:27,506 --> 02:12:29,141
VARY SLIGHTLY IN STRUCTURE AMONG

2667
02:12:29,141 --> 02:12:36,816
THEMES SELVES AND MAY DEVIATE

2668
02:12:36,816 --> 02:12:38,250
FROM ACT ICOSAHEDRAL SYMMETRY.

2669
02:12:38,250 --> 02:12:43,956
IF YOU REMOVE THE GLYCOPROTEINS,

2670
02:12:43,956 --> 02:12:48,994
ISOLATE BEFORE THEY TARGET

2671
02:12:48,994 --> 02:12:50,429
GLYCOPROTEINS HOW THAT STRUCTURE

2672
02:12:50,429 --> 02:12:52,698
IS NOT SYMMETRIC?

2673
02:12:52,698 --> 02:12:54,900
SHE SHOWED A FEW YEARS LATER

2674
02:12:54,900 --> 02:12:59,171
PARTICLES ARE NOT EVENLY CLOSE

2675
02:12:59,171 --> 02:13:01,874
TO RESEMBLE SPEARICAL PARTICLES.

2676
02:13:01,874 --> 02:13:04,643
HIV MAKES A LOT OF MISTAKES, I

2677
02:13:04,643 --> 02:13:07,279
WONDER ARE THESE VIRUSES MAKING

2678
02:13:07,279 --> 02:13:08,147
MISTAKES LIKE HIV?

2679
02:13:08,147 --> 02:13:12,318
SO HERE IS A CORE WE'RE

2680
02:13:12,318 --> 02:13:13,285
STUDYING.

2681
02:13:13,285 --> 02:13:16,021
WE'RE ASSUMING FIRST IN HEXAMERS

2682
02:13:16,021 --> 02:13:26,465
AND PENTAMERS, THERE ARE A

2683
02:13:27,399 --> 02:13:29,268
COUPLE INTERACTIONS, INTERNAL

2684
02:13:29,268 --> 02:13:37,142
INTERACTIONS, AND IN THE HEXAMER

2685
02:13:37,142 --> 02:13:38,110
BECAUSE IT'S ASYMMETRIC, THE

2686
02:13:38,110 --> 02:13:39,879
INTERACTION IS NOT AT THE SAME

2687
02:13:39,879 --> 02:13:40,312
DISTANCE AS THIS.

2688
02:13:40,312 --> 02:13:42,748
WHEN YOU LOOK AT THE PENTAMER,

2689
02:13:42,748 --> 02:13:44,183
SAME PRINCIPLE, YOU SEE

2690
02:13:44,183 --> 02:13:45,584
INTERACTIONS ARE SIMILAR TO EACH

2691
02:13:45,584 --> 02:13:49,321
OTHER AND AGAIN THE SAME BASIC

2692
02:13:49,321 --> 02:13:55,127
AND AMINO ACIDS SEEM TO BE

2693
02:13:55,127 --> 02:13:57,096
PARTICIPATING IN THE ASSEMBLY OF

2694
02:13:57,096 --> 02:14:02,701
THE CAPSOMER, WE WANTED TO KNOW

2695
02:14:02,701 --> 02:14:05,137
IF THESE -- WHICH IT MAKES

2696
02:14:05,137 --> 02:14:08,040
SENSE, WE WANT SOMETHING STABLE,

2697
02:14:08,040 --> 02:14:10,976
NEUTRAL PH, BUT IT CAN FALL

2698
02:14:10,976 --> 02:14:14,280
APART WHEN IT BECOMES ACIDIC.

2699
02:14:14,280 --> 02:14:16,348
OUR IDEA, BECAUSE ALAN SAID IT

2700
02:14:16,348 --> 02:14:21,720
RIGHT, I WAS A PHYSICIST, FISCAL

2701
02:14:21,720 --> 02:14:26,292
'DEMIST, WE WANTED TO STUDY

2702
02:14:26,292 --> 02:14:30,029
INTERACTIONS RESPONSIBLE FOR

2703
02:14:30,029 --> 02:14:31,764
ASSEMBLY OF CAPSOMER.

2704
02:14:31,764 --> 02:14:40,673
BEFORE DOING MUTATIONS, WE

2705
02:14:40,673 --> 02:14:45,444
LOOKED AT OVER 1800 UNIQUE

2706
02:14:45,444 --> 02:14:49,148
SEQUENCES AND THIS IS AN RNA

2707
02:14:49,148 --> 02:14:50,983
VIRUS, QUITE SURPRISING,

2708
02:14:50,983 --> 02:14:53,118
CONSERVED IN 99.4% OF ALL

2709
02:14:53,118 --> 02:14:54,386
SEQUENCES FOR PATIENTS.

2710
02:14:54,386 --> 02:14:56,922
SO I THOUGHT, OKAY, THAT'S FINE.

2711
02:14:56,922 --> 02:14:57,589
HIGHLY CONSERVED.

2712
02:14:57,589 --> 02:15:01,527
THAT'S A GOOD STARTING POINT TO

2713
02:15:01,527 --> 02:15:02,428
DOING MUTATIONS.

2714
02:15:02,428 --> 02:15:05,130
BUT I'M ALWAYS SECOND GUESSING

2715
02:15:05,130 --> 02:15:05,798
MYSELF.

2716
02:15:05,798 --> 02:15:07,266
AND THEN, OKAY, WAIT, BEFORE

2717
02:15:07,266 --> 02:15:09,668
DOING ANY MUTATION LOOK AT

2718
02:15:09,668 --> 02:15:13,839
DIFFERENT ALPHAVIRUSES, THIS IS

2719
02:15:13,839 --> 02:15:22,781
JUST A LIST OF PHYLOGENETIC

2720
02:15:22,781 --> 02:15:23,482
TREE.

2721
02:15:23,482 --> 02:15:24,650
CryoEM OR X-RAY STRUCTURES.

2722
02:15:24,650 --> 02:15:27,953
LEFT-HAND SIDE IS ONE OF THE

2723
02:15:27,953 --> 02:15:29,688
CryoEM STRUCTURES, CAPSID

2724
02:15:29,688 --> 02:15:30,889
PROTEIN, I JUST HIGHLIGHTED FOUR

2725
02:15:30,889 --> 02:15:32,191
AMINO ACIDS I'M TALKING ABOUT

2726
02:15:32,191 --> 02:15:32,524
TODAY.

2727
02:15:32,524 --> 02:15:33,959
THERE ARE ANOTHER FOUR THAT HAVE

2728
02:15:33,959 --> 02:15:35,594
SOMETHING ELSE TO DO BUT I WON'T

2729
02:15:35,594 --> 02:15:36,695
TALK ABOUT TODAY.

2730
02:15:36,695 --> 02:15:39,198
AND THEN ON THE RIGHT IT'S AN

2731
02:15:39,198 --> 02:15:43,369
OVERLAP OF THE STRUCTURAL FOR

2732
02:15:43,369 --> 02:15:46,672
THE CAPSID PROTEIN OF CHIKV, AND

2733
02:15:46,672 --> 02:15:49,074
THEY ARE CLOSE TO EACH OTHER

2734
02:15:49,074 --> 02:15:50,809
GENETICALLY, THERE'S A

2735
02:15:50,809 --> 02:15:52,344
PERFECT -- ALMOST PERFECT MATCH

2736
02:15:52,344 --> 02:15:53,112
IN STRUCTURE.

2737
02:15:53,112 --> 02:15:55,647
WHEN YOU SEE BLUE AMINO ACID YOU

2738
02:15:55,647 --> 02:15:57,816
SEE BLUE AMINO ACID OR REGULAR

2739
02:15:57,816 --> 02:15:58,584
RED.

2740
02:15:58,584 --> 02:16:01,186
NOW WE HAVE STRUCTURE FOR EIGHT

2741
02:16:01,186 --> 02:16:04,823
OR NINE -- I FORGET HOW MANY

2742
02:16:04,823 --> 02:16:06,658
ALPHAVIRUSES.

2743
02:16:06,658 --> 02:16:08,594
WE USE ALPHAFOLD 2 AND 3 TO

2744
02:16:08,594 --> 02:16:11,163
PREDICT THE REST OF THE

2745
02:16:11,163 --> 02:16:15,968
STRUCTURE, ONE OF THE MOST

2746
02:16:15,968 --> 02:16:17,469
DISTANT ALPHAVIRUSES, AND

2747
02:16:17,469 --> 02:16:22,875
THERE'S SOME CHANGES FOR EXAMPLE

2748
02:16:22,875 --> 02:16:25,411
LYSINE, HERE HIGHLY CONSERVED IN

2749
02:16:25,411 --> 02:16:28,714
OLD WORLD ALPHAVIRUSES, NOW

2750
02:16:28,714 --> 02:16:30,215
INSTEAD OF BEING POSITIVE, IT'S

2751
02:16:30,215 --> 02:16:30,649
NEGATIVE.

2752
02:16:30,649 --> 02:16:33,886
WE START USING THIS APPROACH OF

2753
02:16:33,886 --> 02:16:34,820
MATCHING STRUCTURES, BECAUSE IF

2754
02:16:34,820 --> 02:16:38,924
YOU LOOK AT THE PRIMARY SEQUENCE

2755
02:16:38,924 --> 02:16:40,192
OF CAPSID PROTEINS THEY ARE

2756
02:16:40,192 --> 02:16:44,229
DIFFERENT FROM EACH OTHER.

2757
02:16:44,229 --> 02:16:46,098
WE STARTED OVERLAPPING ALL THE

2758
02:16:46,098 --> 02:16:47,399
STRUCTURES SO WE COULD USE THEM

2759
02:16:47,399 --> 02:16:52,571
AS PINPOINTS TO BE ABLE TO DO A

2760
02:16:52,571 --> 02:16:53,439
PHYLOGENETIC ANALYSIS THAT I

2761
02:16:53,439 --> 02:16:55,307
WON'T TALK ABOUT TODAY.

2762
02:16:55,307 --> 02:16:56,608
ALPHAVIRUSS, YOU CAN DIVIDE THEM

2763
02:16:56,608 --> 02:17:00,679
AS OLD WORLD OR NEW WORLD.

2764
02:17:00,679 --> 02:17:05,050
FOR THOSE WHO WE KNOW THE

2765
02:17:05,050 --> 02:17:07,553
MAMMALIAN HOST, BY MOSQUITOES,

2766
02:17:07,553 --> 02:17:08,654
INVERTEBRATES, VERTEBRATES.

2767
02:17:08,654 --> 02:17:11,723
OLD WORLD MEANS ANY ALPHAVIRUS

2768
02:17:11,723 --> 02:17:13,358
THAT FITS THE WINDOW OF

2769
02:17:13,358 --> 02:17:14,660
MAMMALIAN HOST.

2770
02:17:14,660 --> 02:17:17,930
THAT IS FOUND MANY CONTINENT

2771
02:17:17,930 --> 02:17:21,233
EXCEPT AMERICAS, NEW WORLD JUST

2772
02:17:21,233 --> 02:17:22,201
RESTRICTED TO AMERICAS.

2773
02:17:22,201 --> 02:17:25,804
WE HAVE A FEW ALPHAVIRUSES FOUND

2774
02:17:25,804 --> 02:17:28,440
IN MOSQUITOES, NO IDEA THE

2775
02:17:28,440 --> 02:17:32,911
VERTEBRATE HOST. 

2776
02:17:32,911 --> 02:17:33,912
ALSO ALPHAVIRUSES THAT AFFECT

2777
02:17:33,912 --> 02:17:36,415
FISH AND SEA LIONS.

2778
02:17:36,415 --> 02:17:39,818
WHAT'S SURPRISING FOR OLD WORLD,

2779
02:17:39,818 --> 02:17:41,453
100 CONSERVED, DOESN'T MATTER IF

2780
02:17:41,453 --> 02:17:46,058
THE VIRUS IS FROM AFRICA OR

2781
02:17:46,058 --> 02:17:48,260
NETHERLANDS OR NEW ZEALAND,

2782
02:17:48,260 --> 02:17:50,095
THESE FOUR AMINO ACIDS ARE

2783
02:17:50,095 --> 02:17:50,462
CONSERVED.

2784
02:17:50,462 --> 02:17:52,965
IF YOU TAKE NEW WORLD

2785
02:17:52,965 --> 02:17:57,636
ALPHAVIRUS, IF TAMPA, AND FROM

2786
02:17:57,636 --> 02:18:03,575
VENEZUELA, THEN THESE ARE

2787
02:18:03,575 --> 02:18:06,445
INTERACTIONS OF THE CAPSOMER NOT

2788
02:18:06,445 --> 02:18:06,879
CONSERVED.

2789
02:18:06,879 --> 02:18:09,581
THOSE WHO DON'T KNOW THE

2790
02:18:09,581 --> 02:18:14,853
VERTEBRATE HOST, THREE OF FOUR

2791
02:18:14,853 --> 02:18:16,188
CON SEARCHED.

2792
02:18:16,188 --> 02:18:20,225
SUGGESTING STRONG EVOLUTIONARY

2793
02:18:20,225 --> 02:18:23,195
PRESSURE ON INTERACTIONS, WHY

2794
02:18:23,195 --> 02:18:26,598
ARE NOT INNER INTERACTIONS

2795
02:18:26,598 --> 02:18:27,900
HIGHLY CONSERVED?

2796
02:18:27,900 --> 02:18:30,068
WE FOUND HIGHLY AMOUNT OF

2797
02:18:30,068 --> 02:18:34,339
CONSERVATION, THEN I WAS BRAVE

2798
02:18:34,339 --> 02:18:37,743
ENOUGH TO START DOING

2799
02:18:37,743 --> 02:18:43,115
MUTAGENESIS THANKS TO MITCH IN

2800
02:18:43,115 --> 02:18:48,253
ALAN'S LAB, HEAD OF MOLECULAR

2801
02:18:48,253 --> 02:18:50,222
CLONING.

2802
02:18:50,222 --> 02:18:52,090
WE CONSTRUCTED A PLASMATE, WE

2803
02:18:52,090 --> 02:18:54,826
WAIT 24 HOURS, WATCH THEM WELL,

2804
02:18:54,826 --> 02:18:58,263
WAIT ANOTHER 24 HOURS, TAKE THE

2805
02:18:58,263 --> 02:19:00,132
SUPERNATANT AND FREEZE IT, TAKE

2806
02:19:00,132 --> 02:19:03,235
IT AND INFECT NAIVE CELLS, DO

2807
02:19:03,235 --> 02:19:05,370
THESE IN 48 WELL PLATES.

2808
02:19:05,370 --> 02:19:08,974
AND WE'RE LUCKY TO HAVE THESE

2809
02:19:08,974 --> 02:19:10,943
AUTOMATIC MICROSCOPES BECAUSE

2810
02:19:10,943 --> 02:19:12,044
IT'S FLUORESCENTS MICROSCOPE,

2811
02:19:12,044 --> 02:19:16,315
BECAUSE MUCH FLUORESCENT GENE WE

2812
02:19:16,315 --> 02:19:22,788
CAN DO SEVERAL RAL REPLICATES FR

2813
02:19:22,788 --> 02:19:24,189
EACH EXPERIMENT.

2814
02:19:24,189 --> 02:19:26,491
WHATEVER WE FIND INTERESTING WE

2815
02:19:26,491 --> 02:19:34,499
TAKE TO ELECTRON MICROSCOPE.

2816
02:19:34,499 --> 02:19:36,468
SO THIS IS INTERACTION BETWEEN

2817
02:19:36,468 --> 02:19:41,106
AMINO ACID HERE CONSERVED IN

2818
02:19:41,106 --> 02:19:46,211
93%, THIS ONE 98%, WE CHANGE

2819
02:19:46,211 --> 02:19:50,582
BASIC AMINO ACID OR REVERSE THE

2820
02:19:50,582 --> 02:19:52,884
CHARGES SO THERE'S NO

2821
02:19:52,884 --> 02:19:54,319
ELECTROSTATIC REPULSION BUT NOW

2822
02:19:54,319 --> 02:19:57,256
DON'T HAVE POSITIVE AND NEGATIVE

2823
02:19:57,256 --> 02:19:57,522
INTERFACE.

2824
02:19:57,522 --> 02:20:02,060
ALL THIS WORK HAS BEEN DONE SO

2825
02:20:02,060 --> 02:20:12,604
FAR, MOST BY UNDERGRAD STUDENTS.

2826
02:20:16,174 --> 02:20:17,976
IT WAS VERY INTERESTING BECAUSE

2827
02:20:17,976 --> 02:20:20,045
HERE FOR THE MUTANTS YOU SEE

2828
02:20:20,045 --> 02:20:23,115
REPULSION, SO THAT MAKES SENSE.

2829
02:20:23,115 --> 02:20:24,416
HERE YOU SEE REPULSION.

2830
02:20:24,416 --> 02:20:25,284
MAKES SENSE.

2831
02:20:25,284 --> 02:20:29,354
BUT HERE THERE'S NO REPULSION

2832
02:20:29,354 --> 02:20:31,423
BUT NOW WE DESTROY ABILITY OF

2833
02:20:31,423 --> 02:20:40,732
THE CAPSID PROTEIN TO BEHAVE

2834
02:20:40,732 --> 02:20:43,669
LIKE A DIPOLS, DEPENDS ON THE

2835
02:20:43,669 --> 02:20:44,970
ANGLE, VERY IMPORTANT FOR PART

2836
02:20:44,970 --> 02:20:46,305
OF THE STORY.

2837
02:20:46,305 --> 02:20:48,807
THEN WE SAY, OKAY, WHAT'S WRONG

2838
02:20:48,807 --> 02:20:49,808
WITH THESE VIRUSES?

2839
02:20:49,808 --> 02:20:52,010
WITHIN THE LAB, THANKS TO

2840
02:20:52,010 --> 02:20:58,283
TRAINING I RECEIVED AT NCI.

2841
02:20:58,283 --> 02:21:03,088
YOU CAN SEE BETWEEN TWO CELLS,

2842
02:21:03,088 --> 02:21:03,855
BUDDING PARTICLES.

2843
02:21:03,855 --> 02:21:06,591
AND THE COOL THING ABOUT ALPHA

2844
02:21:06,591 --> 02:21:10,963
VIRUSES, HOW CAN I FIND A CELL

2845
02:21:10,963 --> 02:21:12,264
THAT IS INFECTED?

2846
02:21:12,264 --> 02:21:19,037
THINGS IN THE CIRCLES ARE TYPE 2

2847
02:21:19,037 --> 02:21:21,239
WHERE THE GENOME IS REPLICATED,

2848
02:21:21,239 --> 02:21:26,178
ISOLATED FROM THE REST OF THE

2849
02:21:26,178 --> 02:21:28,146
CELL, SO CGAS CANNOT DETECT OR

2850
02:21:28,146 --> 02:21:30,882
STING, I DON'T REMEMBER, DOESN'T

2851
02:21:30,882 --> 02:21:32,851
MATTER, BUT FOUND THE VIRAL RNA.

2852
02:21:32,851 --> 02:21:36,121
IN THIS USEFUL BECAUSE WE SEE OR

2853
02:21:36,121 --> 02:21:38,323
LOOK FOR CELLS THAT HAVE THESE

2854
02:21:38,323 --> 02:21:41,159
REPLICATION SITES AND WE LOOK TO

2855
02:21:41,159 --> 02:21:43,562
THE -- MOVE AND LOOK TO THE

2856
02:21:43,562 --> 02:21:45,197
PLASMA MEMBRANE TO SEE IF WE CAN

2857
02:21:45,197 --> 02:21:46,431
FIND VIRUSES.

2858
02:21:46,431 --> 02:21:49,267
FOR THE MUTANTS WHERE WE CHANGE

2859
02:21:49,267 --> 02:21:52,871
POSITIVE FOR NEGATIVE AMINO

2860
02:21:52,871 --> 02:21:54,272
ACID, OUTER INTERACTION, WE HAD

2861
02:21:54,272 --> 02:21:58,810
TO LOOK FOR A LOT OF CELLS UNTIL

2862
02:21:58,810 --> 02:22:02,614
WE FOUND CYTOPATHIC VESICLES,

2863
02:22:02,614 --> 02:22:04,716
THIS IS REALLY BEAUTIFUL.

2864
02:22:04,716 --> 02:22:09,321
LARSON IN SWEDEN HAS DONE

2865
02:22:09,321 --> 02:22:10,222
BEAUTIFUL WORK.

2866
02:22:10,222 --> 02:22:11,623
OUR SURPRISE WE FOUND TUBES,

2867
02:22:11,623 --> 02:22:12,524
LONG TUBES.

2868
02:22:12,524 --> 02:22:16,228
THESE TUBES ARE NOT PRESENT IN

2869
02:22:16,228 --> 02:22:17,963
UNINFECTED CELLS, OR EVEN

2870
02:22:17,963 --> 02:22:20,499
INFECTED CELL WITH WILD TYPE,

2871
02:22:20,499 --> 02:22:22,467
TUBES ARE REALLY COOL BECAUSE --

2872
02:22:22,467 --> 02:22:29,341
THIS GOES BACK TO CASPER, WELL,

2873
02:22:29,341 --> 02:22:36,281
GOES BACK TO IF YOU WANT TO MAKE

2874
02:22:36,281 --> 02:22:42,053
THIS, YOU ONLY NEED 12 PENTERS

2875
02:22:42,053 --> 02:22:42,554
PENTAMERS.

2876
02:22:42,554 --> 02:22:46,758
IF YOU WANT TO MAKE LARGER YOU

2877
02:22:46,758 --> 02:22:48,026
NEED HEXAMERS, A DISCRETE

2878
02:22:48,026 --> 02:22:50,662
NUMBER, I WON'T DESCRIBE HOW

2879
02:22:50,662 --> 02:22:51,263
THAT HAPPENS.

2880
02:22:51,263 --> 02:22:54,966
IF YOU ONLY HAVE HEXAMERS YOU

2881
02:22:54,966 --> 02:22:57,803
GET TWO POSSIBLE STRUCTURES,

2882
02:22:57,803 --> 02:23:00,338
PLANAR SHEETS OR TUBES, LIKE

2883
02:23:00,338 --> 02:23:01,640
CARBON NANOTUBES.

2884
02:23:01,640 --> 02:23:04,843
SO OUR HYPOTHESIS NOW WE'RE

2885
02:23:04,843 --> 02:23:10,949
DOING SIMULATIONS WITH ROY AND

2886
02:23:10,949 --> 02:23:15,654
VICTOR, IN MADRID, TRYING TO SEE

2887
02:23:15,654 --> 02:23:18,723
IF WE CAN SIMULATE ASSEMBLY

2888
02:23:18,723 --> 02:23:20,025
SUGGESTING MUTATION HERE BY

2889
02:23:20,025 --> 02:23:22,561
CHANGING POSITIVE FOR NEGATIVE

2890
02:23:22,561 --> 02:23:23,862
AMINO ACID IS DETABLIZING

2891
02:23:23,862 --> 02:23:30,235
ABILITY OF THE PROTEIN TO FORM

2892
02:23:30,235 --> 02:23:32,304
PENTAMERS BUT NOT HEXAMERS.

2893
02:23:32,304 --> 02:23:37,876
THIS IS A SIMULATION FROM ORAZIO

2894
02:23:37,876 --> 02:23:41,480
VICTOR, ALL SIMULATIONS,

2895
02:23:41,480 --> 02:23:42,347
PRELIMINARY RESULTS.

2896
02:23:42,347 --> 02:23:44,883
AND THE PENTAMER IS NOT -- WILD

2897
02:23:44,883 --> 02:23:47,285
TYPE IS NOT VERY STABLE.

2898
02:23:47,285 --> 02:23:49,454
THAT HAS TO DO WITH ARTIFACTS

2899
02:23:49,454 --> 02:23:52,591
THAT I DON'T HAVE THE TIME TO GO

2900
02:23:52,591 --> 02:23:54,359
OVER THAT.

2901
02:23:54,359 --> 02:23:57,362
WHEN YOU LOOK AT THE MUTANT, IT

2902
02:23:57,362 --> 02:23:58,797
IMMEDIATELY FALLS APART.

2903
02:23:58,797 --> 02:24:02,067
YOU CAN SEE IT FALLING APART,

2904
02:24:02,067 --> 02:24:05,003
CONFIRMING THAT THE PENTAMER

2905
02:24:05,003 --> 02:24:07,439
UNDER THIS SIMULATION IS

2906
02:24:07,439 --> 02:24:08,507
UNSTABLE.

2907
02:24:08,507 --> 02:24:14,312
STILL WAITING FOR HORACI,  TO

2908
02:24:14,312 --> 02:24:15,614
HAVE RESULT FOR HEXAMERS AND

2909
02:24:15,614 --> 02:24:19,017
THEN DO SIMULATION TO UNDERSTAND

2910
02:24:19,017 --> 02:24:20,118
HOW THIS FORMS HEXAMERS.

2911
02:24:20,118 --> 02:24:21,119
IT MAKES SENSE.

2912
02:24:21,119 --> 02:24:22,954
REMEMBER, THIS AMINO ACID IS

2913
02:24:22,954 --> 02:24:23,655
HIGHLY CONSERVED.

2914
02:24:23,655 --> 02:24:26,024
SO THAT'S WHY YOU DON'T SEE

2915
02:24:26,024 --> 02:24:27,325
ANYTIME NATURE, THESE MUTATIONS.

2916
02:24:27,325 --> 02:24:30,061
NOW WHEN WE LOOK AT THE PARTNER

2917
02:24:30,061 --> 02:24:37,068
OF 172 AND CHANGE THAT'S  ACID R

2918
02:24:37,068 --> 02:24:40,005
A BASE, ANOTHER PROJECT WE'RE

2919
02:24:40,005 --> 02:24:42,541
STUDYING BUDDING WITH U.C.

2920
02:24:42,541 --> 02:24:44,943
RIVERSIDE, SO WE HAVE

2921
02:24:44,943 --> 02:24:48,313
CHARACTERIZED BUDDING OF THE

2922
02:24:48,313 --> 02:24:49,414
VIRUS QUITE EXTENSIVELY AND HAVE

2923
02:24:49,414 --> 02:24:53,785
PAPERS SOON TO BE REVIEWED SOON.

2924
02:24:53,785 --> 02:24:59,491
WHAT WE FIND THE CORES STOP AT

2925
02:24:59,491 --> 02:25:04,629
PLASMA MEMBRANE, TURNS OUT THE

2926
02:25:04,629 --> 02:25:06,464
LAB IN SANFORD SHOWED THESE HERE

2927
02:25:06,464 --> 02:25:13,939
AND HERE, IN THE CYTOPLASM, NOT

2928
02:25:13,939 --> 02:25:21,279
FORMING  HEXAGONAL SHEETS.

2929
02:25:21,279 --> 02:25:28,620
WHEN THEY INTERACT THE CORE --

2930
02:25:28,620 --> 02:25:30,055
IT'S WELL KNOWN THAT

2931
02:25:30,055 --> 02:25:34,125
GLYCOPROTEINS IN ORDER TO --

2932
02:25:34,125 --> 02:25:35,527
THEY ARE TOUCHING THE CAPSID

2933
02:25:35,527 --> 02:25:39,497
PROTEIN AND CORE VERY STRONGLY,

2934
02:25:39,497 --> 02:25:42,968
THEY NEED TO BIND.

2935
02:25:42,968 --> 02:25:46,705
SO WHILE WE'RE ASSUMING, THESE

2936
02:25:46,705 --> 02:25:56,114
CORES THAT WE'RE FORMING CANNOT

2937
02:25:56,114 --> 02:25:57,215
REANEAL THEMSELVES.

2938
02:25:57,215 --> 02:26:01,720
WE LOOK AT SOME MUTATIONS IN THE

2939
02:26:01,720 --> 02:26:04,689
PENTAMERS AND HEXAMERS, STILL

2940
02:26:04,689 --> 02:26:07,525
MISSING A FEW MUTATIONS, LYSINE

2941
02:26:07,525 --> 02:26:09,728
TO USINE IS NOT BAD, WE FIND

2942
02:26:09,728 --> 02:26:14,766
THIS IN NEW WORLD ALPHAVIRUSES,

2943
02:26:14,766 --> 02:26:16,501
SURPRISED CHANGING BASIC FOR

2944
02:26:16,501 --> 02:26:18,470
ACIDIC WAS NOT THAT BAD.

2945
02:26:18,470 --> 02:26:22,974
BUT IF YOU LOOK, IF WE DO THE

2946
02:26:22,974 --> 02:26:27,345
OPPOSITE CHANGE IN 184, CHANGE

2947
02:26:27,345 --> 02:26:28,747
ACID FOR HYDROPHOBIC, COULD BE

2948
02:26:28,747 --> 02:26:30,949
LOSING A PROTEIN, THE VIRUS IS

2949
02:26:30,949 --> 02:26:31,916
PRETTY MUCH DEAD.

2950
02:26:31,916 --> 02:26:33,351
SEEMS TO ME THAT ONE AMINO ACID

2951
02:26:33,351 --> 02:26:35,086
IS MORE CRITICAL THAN THE OTHER,

2952
02:26:35,086 --> 02:26:42,661
AND WE HAVE NO IDEA WHAT THAT

2953
02:26:42,661 --> 02:26:42,994
MEANS.

2954
02:26:42,994 --> 02:26:44,529
SO, WE'RE THINKING OF WHAT'S

2955
02:26:44,529 --> 02:26:47,298
GOING ON, AND WE SAY, WELL, THIS

2956
02:26:47,298 --> 02:26:48,900
IS A PHYSICAL CHEMISTRY PROJECT,

2957
02:26:48,900 --> 02:26:50,435
BUT THESE ARE VIRUSES SO LET'S

2958
02:26:50,435 --> 02:26:51,870
THINK ABOUT THE HOST.

2959
02:26:51,870 --> 02:27:00,812
IT TURNS OUT THE OLD WORLD

2960
02:27:00,812 --> 02:27:02,247
ALPHAVIRUSES ARE NON-LETHAL.

2961
02:27:02,247 --> 02:27:04,549
THE DISEASE IS NASTY BUT YOU

2962
02:27:04,549 --> 02:27:06,851
WON'T DIE FROM CHIKUNGUNYA.

2963
02:27:06,851 --> 02:27:08,286
IF YOU'RE INFECTED, A HORSE

2964
02:27:08,286 --> 02:27:10,255
INFECTED WITH A NEW WORLD

2965
02:27:10,255 --> 02:27:13,425
ALPHAVIRUS, YOU MIGHT DIE MOST

2966
02:27:13,425 --> 02:27:15,493
LIKELY FROM ENCEPHALITIS, HIGHLY

2967
02:27:15,493 --> 02:27:15,727
LETHAL.

2968
02:27:15,727 --> 02:27:18,997
THEY ARE DIFFERENT IN THE WAY

2969
02:27:18,997 --> 02:27:21,633
THEY BEHAVE IN MAMMALS.

2970
02:27:21,633 --> 02:27:30,408
SO A COUPLE YEARS AGO KAEBLER AT

2971
02:27:30,408 --> 02:27:33,244
GUTTERS ARE DID CryoEM OF

2972
02:27:33,244 --> 02:27:36,748
CHIKUNGUNYA, AND HE WAS AN

2973
02:27:36,748 --> 02:27:40,819
HONEST MICROSCOPIST. 

2974
02:27:40,819 --> 02:27:41,386
BEFORE DOING RECONSTRUCTION

2975
02:27:41,386 --> 02:27:42,754
REALIZED THERE WERE FOUR

2976
02:27:42,754 --> 02:27:44,422
POPULATIONS OF PARTICLES, IN

2977
02:27:44,422 --> 02:27:48,026
BLACK NORMAL SIZE, THEN SMALLER

2978
02:27:48,026 --> 02:27:52,197
ONE, ONE IN YELLOW, LARGER ONES

2979
02:27:52,197 --> 02:27:54,499
IN GREEN, AND MULTI-CORE IN RED.

2980
02:27:54,499 --> 02:27:57,235
TURNS OUT HE MADE THESE

2981
02:27:57,235 --> 02:27:59,104
INTERESTING OBSERVATIONS THAT HE

2982
02:27:59,104 --> 02:28:01,706
HASN'T TAKEN FURTHER FROM THIS

2983
02:28:01,706 --> 02:28:03,942
TABLE THAT CHIKUNGUNYA MAKES A

2984
02:28:03,942 --> 02:28:07,846
MISTAKE IN 4% OF THE PARTICLES

2985
02:28:07,846 --> 02:28:11,883
BUT VEEV HIGHLY LETHAL IN TEN

2986
02:28:11,883 --> 02:28:12,784
AND EEEV IN 12%.

2987
02:28:12,784 --> 02:28:17,155
WHEN YOU LOOK AT INTERACTIONS,

2988
02:28:17,155 --> 02:28:21,192
YOU SEE EEEV INSTEAD OF HAVING

2989
02:28:21,192 --> 02:28:23,261
STRONG ELECTROSTATIC

2990
02:28:23,261 --> 02:28:24,362
INTERACTIONS NOW YOU CHARGE

2991
02:28:24,362 --> 02:28:32,370
POLAR AMINO ACID FOR

2992
02:28:32,370 --> 02:28:34,005
NON-POLAR -- SORRY, NON-POLAR,

2993
02:28:34,005 --> 02:28:36,741
NOW HAVE YOU TWO HYDROPHOBIC

2994
02:28:36,741 --> 02:28:37,275
INTERACTIONS.

2995
02:28:37,275 --> 02:28:40,011
WE HAVE PROPOSED, WELL, RECENTLY

2996
02:28:40,011 --> 02:28:42,080
POSTED THIS QUESTION IN JOURNAL

2997
02:28:42,080 --> 02:28:46,251
OF VIROLOGY THAT ARE MISTAKES IN

2998
02:28:46,251 --> 02:28:50,255
ASSEMBLY A WAY TO ATTENUATE

2999
02:28:50,255 --> 02:28:51,189
HIGHLY PATHOGENIC ALPHAVIRUSES

3000
02:28:51,189 --> 02:28:54,125
AND IS INTERACTION BETWEEN TWO

3001
02:28:54,125 --> 02:28:56,428
AMINO ACIDS UPON FOR FIDELITY OF

3002
02:28:56,428 --> 02:28:57,595
ASSEMBLY PROCESS OR DO WE NEED

3003
02:28:57,595 --> 02:29:01,466
TO FIND MORE AMINO ACIDS?

3004
02:29:01,466 --> 02:29:02,467
AND WE'RE LOOKING AT ANOTHER

3005
02:29:02,467 --> 02:29:03,868
FOUR SET OF AMINO ACIDS BUT I

3006
02:29:03,868 --> 02:29:06,371
DON'T HAVE ENOUGH TIME TO TELL

3007
02:29:06,371 --> 02:29:08,039
YOU ABOUT THOSE.

3008
02:29:08,039 --> 02:29:09,974
SO JUST TO FINISH HOPEFULLY ON

3009
02:29:09,974 --> 02:29:15,146
TIME, WE HAVE FOUND HIGHLY

3010
02:29:15,146 --> 02:29:16,948
CONSERVED -- THAT THESE ARE

3011
02:29:16,948 --> 02:29:19,083
HIGHLY CONSERVED AND PRETTY MUCH

3012
02:29:19,083 --> 02:29:20,485
ANY MUTATIONS HERE KILL THE

3013
02:29:20,485 --> 02:29:22,987
VIRUSES, I SHOWED THREE BUT WE

3014
02:29:22,987 --> 02:29:24,322
HAVE MORE.

3015
02:29:24,322 --> 02:29:26,825
THAT THE INNER INTERACTIONS ARE

3016
02:29:26,825 --> 02:29:28,693
ONLY CONSERVED OLD WORLD

3017
02:29:28,693 --> 02:29:31,896
ALPHAVIRUSES, NOT CONSERVED IN

3018
02:29:31,896 --> 02:29:33,865
OTHER ALPHAVIRUSES SO THE VIRUS

3019
02:29:33,865 --> 02:29:35,099
IS TOLERANT TO MUTATIONS HERE,

3020
02:29:35,099 --> 02:29:37,035
NOT TO ALL OF THEM.

3021
02:29:37,035 --> 02:29:39,437
IN FACT, THEY ARE NOT CONSERVED

3022
02:29:39,437 --> 02:29:41,840
THE SAME WAY.

3023
02:29:41,840 --> 02:29:47,545
184 IS MORE CONSERVED THAN 177,

3024
02:29:47,545 --> 02:29:50,281
AND 184 IS LESS TOLERANT TO

3025
02:29:50,281 --> 02:29:51,149
MUTATION THAN 177.

3026
02:29:51,149 --> 02:29:52,617
RELATION BETWEEN DEGREE OF

3027
02:29:52,617 --> 02:29:54,652
CONSERVATION OF THESE AMINO

3028
02:29:54,652 --> 02:29:56,187
ACIDS IN NATURE AND WHAT ARE WE

3029
02:29:56,187 --> 02:29:57,489
DOING IN THE LAB.

3030
02:29:57,489 --> 02:30:07,365
AND THE FINAL QUESTION THAT IS

3031
02:30:07,365 --> 02:30:09,100
REALLY PUZZLING, ARE MAY TAKES

3032
02:30:09,100 --> 02:30:14,305
IN ASSEMBLY A WAY TO ATTENUATE

3033
02:30:14,305 --> 02:30:19,944
HIGHLY PATHOGENIC VIRUS, NOW

3034
02:30:19,944 --> 02:30:20,945
CHIKUNGUNYA, ALSO MUTATING VEEV

3035
02:30:20,945 --> 02:30:24,449
TO MAKE IT LOOK LIKE  CHIKUNGUNA

3036
02:30:24,449 --> 02:30:27,952
BUT ONLY THE CAPSID PROTEIN TO

3037
02:30:27,952 --> 02:30:38,363
SEE IF WE CAN PROVE THE

3038
02:30:40,064 --> 02:30:40,565
HYPOTHESIS.

3039
02:30:40,565 --> 02:30:45,136
THESE ARE THE PEOPLE HERE.

3040
02:30:45,136 --> 02:30:47,672
CAROLINA, MIGUEL, WORK DONE IN

3041
02:30:47,672 --> 02:30:50,642
PART WITH JUAN AND ROYA AND

3042
02:30:50,642 --> 02:30:51,175
VICTOR.

3043
02:30:51,175 --> 02:30:54,612
THIS IS FUNDING AGENCIES THAT

3044
02:30:54,612 --> 02:30:55,813
SUPPORT OUR PROJECTS.

3045
02:30:55,813 --> 02:30:58,883
I HAVE TO THANK YOU FOR

3046
02:30:58,883 --> 02:31:00,351
EVERYTHING, ALAN.

3047
02:31:00,351 --> 02:31:01,686
GOOD LUCK.

3048
02:31:01,686 --> 02:31:04,789
AND I COULDN'T -- I COULDN'T

3049
02:31:04,789 --> 02:31:06,257
HAVE MORE LUCK BECAUSE WHEN I

3050
02:31:06,257 --> 02:31:09,227
MET YOU MY LIFE REALLY, REALLY

3051
02:31:09,227 --> 02:31:10,662
CHANGED.

3052
02:31:10,662 --> 02:31:13,831
AND I HOPE I HAD -- I WAS NOT

3053
02:31:13,831 --> 02:31:14,365
OVER TIME.

3054
02:31:14,365 --> 02:31:16,200
AND THANK YOU VERY MUCH.

3055
02:31:16,200 --> 02:31:17,769
I DON'T KNOW IF YOU HAVE ANY

3056
02:31:17,769 --> 02:31:19,971
QUESTIONS.

3057
02:31:19,971 --> 02:31:25,977
 

3058
02:31:25,977 --> 02:31:29,714
 

3059
02:31:29,714 --> 02:31:30,682
>> CALL OUT IF YOU HAVE

3060
02:31:30,682 --> 02:31:32,884
QUESTIONS.

3061
02:31:32,884 --> 02:31:39,590
 

3062
02:31:39,590 --> 02:31:44,929
 

3063
02:31:44,929 --> 02:31:45,129
YEAH.

3064
02:31:45,129 --> 02:31:49,334
>> NICE TALK.

3065
02:31:49,334 --> 02:31:55,673
WE FOUND THAT DEFECTS CAN

3066
02:31:55,673 --> 02:31:59,610
FACILITATE DISASSOCIATION.

3067
02:31:59,610 --> 02:32:01,245
SO, ARE YOU SEEING YOUR

3068
02:32:01,245 --> 02:32:03,014
PARTICLES WITH LESSER ABILITY TO

3069
02:32:03,014 --> 02:32:08,453
ASSEMBLY ALSO ABLE TO

3070
02:32:08,453 --> 02:32:09,354
DISASSEMBLE MORE READILY.

3071
02:32:09,354 --> 02:32:10,321
>> GOOD QUESTION.

3072
02:32:10,321 --> 02:32:11,623
WHEN THEY HAVE PROTEINS, NO.

3073
02:32:11,623 --> 02:32:13,791
WE HAVE DONE LOTS OF

3074
02:32:13,791 --> 02:32:14,092
EXPERIMENTS.

3075
02:32:14,092 --> 02:32:17,595
BUT WHEN WE REMOVE THE

3076
02:32:17,595 --> 02:32:19,397
GLYCOPROTEINS, THEN THEY WANT TO

3077
02:32:19,397 --> 02:32:21,933
MAKE LOTS OF MISTAKES, THEY FALL

3078
02:32:21,933 --> 02:32:30,341
APART IMMEDIATELY, THE ONES THAT

3079
02:32:30,341 --> 02:32:33,511
MAKE MISTAKES.

3080
02:32:33,511 --> 02:32:35,513
IT'S MORE ASSEMBLY.

3081
02:32:35,513 --> 02:32:38,783
ONE CAVEAT, WE'RE NOW DOING

3082
02:32:38,783 --> 02:32:39,851
EXPERIMENTS, HOW THEY

3083
02:32:39,851 --> 02:32:40,752
DISASSEMBLE IN THE LYSOSOME

3084
02:32:40,752 --> 02:32:45,456
BECAUSE THEY MIGHT FALL APART

3085
02:32:45,456 --> 02:32:48,226
EITHER IN THE LYSOSOME,

3086
02:32:48,226 --> 02:32:51,696
RELATIVELY HIGHER PACE, COULD BE

3087
02:32:51,696 --> 02:32:57,235
CAUSING LESS INFECTIOUS.

3088
02:32:57,235 --> 02:32:58,036
 

3089
02:32:58,036 --> 02:32:59,203
I DON'T KNOW IF THAT MAKES

3090
02:32:59,203 --> 02:33:02,740
SENSE.

3091
02:33:02,740 --> 02:33:04,709
>> OTHER QUESTIONS, YES?

3092
02:33:04,709 --> 02:33:07,345
>> YEAH, SO I DIDN'T QUITE

3093
02:33:07,345 --> 02:33:10,815
UNDERSTAND WHY THERE'S SO MUCH

3094
02:33:10,815 --> 02:33:12,717
MORE VARIABILITY IN THE CAPSID

3095
02:33:12,717 --> 02:33:15,086
PROTEIN OF THE ALPHAVIRUSES IN

3096
02:33:15,086 --> 02:33:18,956
THE NEW WORLD VIRUSES VERSUS OLD

3097
02:33:18,956 --> 02:33:19,157
WORLD.

3098
02:33:19,157 --> 02:33:21,893
>> OH, SO WE DON'T KNOW.

3099
02:33:21,893 --> 02:33:25,196
WE HAVE FINISHED NOW A

3100
02:33:25,196 --> 02:33:28,466
PHYLOGENETIC ANALYSIS, AND IT IS

3101
02:33:28,466 --> 02:33:28,800
AMAZING.

3102
02:33:28,800 --> 02:33:35,773
ALPHAVIRUSS STARTED AND WE HAVE

3103
02:33:35,773 --> 02:33:37,442
NOW RECONSTRUCTED SEVERAL

3104
02:33:37,442 --> 02:33:39,010
ANCESTORS AND STARTED VARIABLE.

3105
02:33:39,010 --> 02:33:43,448
OUR GUESS WAS AT THE BEGINNING

3106
02:33:43,448 --> 02:33:45,650
HIGHLY PATHOGENIC, AT SOME

3107
02:33:45,650 --> 02:33:49,153
POINT, SOMETHING HAPPENED THAT

3108
02:33:49,153 --> 02:33:51,556
ALPHAVIRUSES BECAME LESS

3109
02:33:51,556 --> 02:33:52,023
PATHOGENIC.

3110
02:33:52,023 --> 02:33:54,625
AND A WAY TO -- OUR HYPOTHESIS

3111
02:33:54,625 --> 02:33:57,028
IS THAT THAT LED THEM TO

3112
02:33:57,028 --> 02:33:58,996
STABILIZE A CAPSID AND THAT

3113
02:33:58,996 --> 02:34:02,266
ALLOWED THEM TO SPLIT ALL

3114
02:34:02,266 --> 02:34:05,136
OVER -- SPREAD ALL OVER THE

3115
02:34:05,136 --> 02:34:05,670
WORLD.

3116
02:34:05,670 --> 02:34:09,173
INTERACTIONS IN PAKISTAN AND

3117
02:34:09,173 --> 02:34:10,842
UGANDA, IT'S UNCANNY, SUSTAINED.

3118
02:34:10,842 --> 02:34:13,444
BUT NOT IN THE AMERICAS.

3119
02:34:13,444 --> 02:34:19,383
AND WE THINK THAT HAS TO DO WITH

3120
02:34:19,383 --> 02:34:21,919
DEGREE OF PATHOGENICITY, THEY

3121
02:34:21,919 --> 02:34:24,222
MADE MISTAKES, IN AMERICA, TO

3122
02:34:24,222 --> 02:34:26,724
HAVE ENOUGH PROBABILITY TO

3123
02:34:26,724 --> 02:34:28,793
INFECT ANOTHER HOST.

3124
02:34:28,793 --> 02:34:34,499
>> SO IS IT HUMAN HOST OR
ARE

3125
02:34:34,499 --> 02:34:35,833
THESE IN MONKEYS, NEW WORLD

3126
02:34:35,833 --> 02:34:36,367
MONKEYS?

3127
02:34:36,367 --> 02:34:39,637
>> OH, IN THE NEW WORLD
IT'S

3128
02:34:39,637 --> 02:34:40,104
MAINLY HORSE.

3129
02:34:40,104 --> 02:34:42,740
HORSES, THIS IS A BIG PROBLEM

3130
02:34:42,740 --> 02:34:45,877
FOR PEOPLE THAT LIKE RACES.

3131
02:34:45,877 --> 02:34:49,514
BUT THEY CAN INFECT MONKEYS.

3132
02:34:49,514 --> 02:34:54,118
THEY CAN INFECT EVEN SEA LIONS.

3133
02:34:54,118 --> 02:35:02,426
TROPISM IS REALLY WIDE. 

3134
02:35:02,426 --> 02:35:03,327
>> THANK YOU.

3135
02:35:03,327 --> 02:35:07,265
>> THANK YOU VERY MUCH. 

3136
02:35:07,265 --> 02:35:08,266
THANK YOU, ALAN.

3137
02:35:08,266 --> 02:35:10,768
>> THANK YOU TO ALL THE
SPEAKERS

3138
02:35:10,768 --> 02:35:12,737
FOR THE MORNING SESSION, REALLY

3139
02:35:12,737 --> 02:35:13,271
FANTASTIC.

3140
02:35:13,271 --> 02:35:14,939
WE'RE ON BREAK UNTIL 2:15.

3141
02:35:14,939 --> 02:35:16,574
THERE'S A CAFETERIA TO YOUR LEFT

3142
02:35:16,574 --> 02:35:18,242
AS YOU LEAVE THE MAIN DOORS

3143
02:35:18,242 --> 02:35:18,442
HERE.

3144
02:35:18,442 --> 02:35:21,872
WE'LL SEE YOU BACK HERE AT 2:15 

3145
02:35:21,872 --> 02:35:24,541
THE NEXT SPEAKER IS A GOOD 

3146
02:35:24,541 --> 02:35:27,011
FRIEND, AN OLD FRIEND, AND ONE 

3147
02:35:27,011 --> 02:35:30,614
OF THE FEW COLLEAGUES WHO 

3148
02:35:30,614 --> 02:35:34,718
PERSISTS IN STUDYING MURINE 

3149
02:35:34,718 --> 02:35:37,321
LEUKEMIA VIRUS DESPITE THE 

3150
02:35:37,321 --> 02:35:38,355
DIFFICULTIES IN GETTING FUNDING 

3151
02:35:38,355 --> 02:35:39,690
FOR A VIRUS THAT DOESN'T INFECT 

3152
02:35:39,690 --> 02:35:42,659
HUMANS.  

3153
02:35:42,659 --> 02:35:50,200
MONICA HAS MADE MANY SEMINAL 

3154
02:35:50,200 --> 02:35:52,136
CONTRIBUTIONS TO MURINE LEUKEMIA

3155
02:35:52,136 --> 02:35:53,704
VIRUS BIOLOGY.  I ALWAYS 

3156
02:35:53,704 --> 02:35:56,073
PARTICULARLY GO BACK TO HER WORK

3157
02:35:56,073 --> 02:36:02,146
ON THE FACT THAT, UNLIKE THE 

3158
02:36:02,146 --> 02:36:04,982
PARADIGM HIV, MURINE LEUKEMIA 

3159
02:36:04,982 --> 02:36:11,288
VIRUS DO NOT ENTER THE NUCLEUS. 

3160
02:36:11,288 --> 02:36:12,423
THEY HOOK UP BY A COMPLETELY 

3161
02:36:12,423 --> 02:36:14,858
DIFFERENT KIND OF MECHANISM.  I 

3162
02:36:14,858 --> 02:36:17,961
FEEL THAT A LOT OF MY WORK 

3163
02:36:17,961 --> 02:36:21,532
RELATES TO THAT BASIC FACT ABOUT

3164
02:36:21,532 --> 02:36:27,704
MLV, WHICH SHE ELUCIDATED.  

3165
02:36:27,704 --> 02:36:29,239
ANYWAY, I DON'T KNOW WHAT SHE'S 

3166
02:36:29,239 --> 02:36:35,712
GOING TO TALK ABOUT TODAY.  HER 

3167
02:36:35,712 --> 02:36:43,554
TITLE IS MYSTERIOUS AND ELUSIVE,

3168
02:36:43,554 --> 02:36:46,023
FROM MLV TO EX MACHINA.  WE'LL 

3169
02:36:46,023 --> 02:36:46,924
FIND OUT WHAT THAT MACHINES.  

3170
02:36:46,924 --> 02:36:57,367
WELCOME, MONICA.  >> SO I 

3171
02:37:02,706 --> 02:37:04,708
DECIDED NOT TO TALK ABOUT MY 

3172
02:37:04,708 --> 02:37:07,010
RESEARCH DIRECTLY AND TALK MORE 

3173
02:37:07,010 --> 02:37:08,345
ABOUT MY COMMUNICATIONS WITH 

3174
02:37:08,345 --> 02:37:10,347
ALAN THROUGH E-MAILS AND THROUGH

3175
02:37:10,347 --> 02:37:11,215
STUDYING MLV.

3176
02:37:11,215 --> 02:37:13,817
I THINK THIS IS PROBABLY THE 

3177
02:37:13,817 --> 02:37:15,652
LAST COLLECTION OF PEOPLE WHO 

3178
02:37:15,652 --> 02:37:17,020
HAVE ACTUALLY WORKED ON THE 

3179
02:37:17,020 --> 02:37:18,822
VIRUS AND STILL MIGHT REMEMBER 

3180
02:37:18,822 --> 02:37:23,727
THE RESTRICTION MAP.

3181
02:37:23,727 --> 02:37:27,798
SO ADD AN AS RESEARCHER, MENTOR,

3182
02:37:27,798 --> 02:37:29,299
FRIEND, AND MOVIE CRITIC 

3183
02:37:29,299 --> 02:37:31,268
BECAUSE, AS YOU'LL SEE, A LOT OF

3184
02:37:31,268 --> 02:37:32,369
THE E-MAILS WERE ABOUT WHAT 

3185
02:37:32,369 --> 02:37:34,738
MOVIES WE'RE WATCHING.

3186
02:37:34,738 --> 02:37:37,040
EVERYONE HAS SEEN THIS TYPE OF 

3187
02:37:37,040 --> 02:37:37,975
PICTURE BEFORE.

3188
02:37:37,975 --> 02:37:40,777
THIS IS A HYBRID BECAUSE I KNOW,

3189
02:37:40,777 --> 02:37:41,912
JONATHAN, THAT WAS A GREAT 

3190
02:37:41,912 --> 02:37:43,881
PICTURE, BUT I KNOW YOU NEVER 

3191
02:37:43,881 --> 02:37:45,983
SAT IN THE FRONT ROW, SO I 

3192
02:37:45,983 --> 02:37:54,958
PLACED YOU IN YOUR CORRECT SPOT.

3193
02:37:54,958 --> 02:37:56,527
IN THE BACK RIGHT SECTION 

3194
02:37:56,527 --> 02:37:56,760
CORNER.

3195
02:37:56,760 --> 02:38:00,230
WE MISS YOU THERE.

3196
02:38:00,230 --> 02:38:01,832
AND STEVE ALSO WAS PLACED IN THE

3197
02:38:01,832 --> 02:38:03,200
FRONT, BUT USUALLY IF YOU WERE 

3198
02:38:03,200 --> 02:38:05,435
LOOKING FOR STEVE, HE'S SORT OF 

3199
02:38:05,435 --> 02:38:06,403
IN THE BACK RIGHT HAND CORNER 

3200
02:38:06,403 --> 02:38:11,308
WHEN HE ACTUALLY SHOWS UP.

3201
02:38:11,308 --> 02:38:12,509
AND OF COURSE, WE HAVE THE FRONT

3202
02:38:12,509 --> 02:38:15,245
ROW PEOPLE, WHO STILL ARE IN THE

3203
02:38:15,245 --> 02:38:17,948
FRONT ROW, BUT IF YOU REALLY 

3204
02:38:17,948 --> 02:38:19,850
WERE LOOKING FOR ALAN, HE REALLY

3205
02:38:19,850 --> 02:38:21,485
WAS IN THAT RIGHT HAND SECTION, 

3206
02:38:21,485 --> 02:38:25,255
A FEW ROWS IN, A FEW ROWS BACK, 

3207
02:38:25,255 --> 02:38:26,957
AND THAT'S WHERE I'VE BEEN 

3208
02:38:26,957 --> 02:38:29,927
SITTING RECENTLY.

3209
02:38:29,927 --> 02:38:31,228
I APOLOGIZE TO EVERYONE ELSE WHO

3210
02:38:31,228 --> 02:38:33,063
ACTUALLY SITS IN OTHER PLACES, 

3211
02:38:33,063 --> 02:38:34,398
BUT IF YOU'RE ON THE LEFT-HAND 

3212
02:38:34,398 --> 02:38:35,799
SIDE OF THE AUDITORIUM, I 

3213
02:38:35,799 --> 02:38:36,400
USUALLY DON'T KNOW WHERE YOU 

3214
02:38:36,400 --> 02:38:40,971
SIT.

3215
02:38:40,971 --> 02:38:42,105
(LAUGHTER) 

3216
02:38:42,105 --> 02:38:44,875
IF YOU'RE LOOKING FOR ALAN IN 

3217
02:38:44,875 --> 02:38:46,243
THE AUDITORIUM, THAT'S WHERE YOU

3218
02:38:46,243 --> 02:38:48,011
WOULD FIND HIM.

3219
02:38:48,011 --> 02:38:49,680
EVERY YEAR I REALLY WOULD -- 

3220
02:38:49,680 --> 02:38:50,581
THIS IS TRUE.

3221
02:38:50,581 --> 02:38:53,884
EVERY YEAR I WOULD GO TO COLD 

3222
02:38:53,884 --> 02:38:55,919
SPRING HARBOR AND SAY TO MYSELF,

3223
02:38:55,919 --> 02:38:59,756
I WONDER WHAT ALAN REIN IS 

3224
02:38:59,756 --> 02:39:00,457
WORKING ON.

3225
02:39:00,457 --> 02:39:02,025
IT WAS ALWAYS SOMETHING NEW AND 

3226
02:39:02,025 --> 02:39:02,559
DIFFERENT.

3227
02:39:02,559 --> 02:39:04,194
IT WASN'T LIKE YOU'RE WORKING ON

3228
02:39:04,194 --> 02:39:05,162
RT ALL THE TIME.

3229
02:39:05,162 --> 02:39:06,563
HE ALWAYS CAME UP WITH SOMETHING

3230
02:39:06,563 --> 02:39:06,830
DIFFERENT.

3231
02:39:06,830 --> 02:39:08,999
AND THE KEY POINT FOR ME IS LIKE

3232
02:39:08,999 --> 02:39:10,367
SOMETIMES, ESPECIALLY IF YOU 

3233
02:39:10,367 --> 02:39:12,436
WORK ON MLV AND YOU SEE A PAPER 

3234
02:39:12,436 --> 02:39:16,106
LIKE THIS AND IS SAYS, OH, 

3235
02:39:16,106 --> 02:39:21,111
GLYCOGAG EFFECTS OF VIRUS ENTRY,

3236
02:39:21,111 --> 02:39:23,146
DEPENDING WHAT SET OF 

3237
02:39:23,146 --> 02:39:24,348
GLYCOPROTEINS YOU WORK ON, IT 

3238
02:39:24,348 --> 02:39:27,050
SENT ME INTO A PANIC BECAUSE I 

3239
02:39:27,050 --> 02:39:30,587
WORK ON GLYCOPROTEIN PARTICLES, 

3240
02:39:30,587 --> 02:39:32,823
AND I WORK ON TARGETING 

3241
02:39:32,823 --> 02:39:35,259
RETROVIRAL PAR I CAN LS BY USING

3242
02:39:35,259 --> 02:39:36,760
DIFFERENT ENVELOPES.

3243
02:39:36,760 --> 02:39:39,129
AND WE USE DIFFERENT PLASMA 

3244
02:39:39,129 --> 02:39:41,598
SYSTEMS, AND I'M SITTING THERE 

3245
02:39:41,598 --> 02:39:44,334
SAYING, OH, MY GOD, WHICH ONE OF

3246
02:39:44,334 --> 02:39:50,274
OUR PLASMA SYSTEMS EXPRESS 

3247
02:39:50,274 --> 02:39:51,341
GLYCOGAG?

3248
02:39:51,341 --> 02:39:54,144
SO HERE ARE TWO FREQUENTLY USED 

3249
02:39:54,144 --> 02:39:55,545
EXPRESSION SYSTEMS.

3250
02:39:55,545 --> 02:39:59,349
ONE WAS FROM THE COSSET LAB, AND

3251
02:39:59,349 --> 02:40:01,985
IT'S CALLED PCEB.

3252
02:40:01,985 --> 02:40:07,491
THE OTHER ONE IS A PCGP FROM THE

3253
02:40:07,491 --> 02:40:09,159
KINGSMEN LAB, AND WHICH ONE OF 

3254
02:40:09,159 --> 02:40:11,128
THESE OR NEITHER OF THESE OR ALL

3255
02:40:11,128 --> 02:40:14,798
OF THEM EXPRESS GLYCOGAG?

3256
02:40:14,798 --> 02:40:15,032
ANYBODY?

3257
02:40:15,032 --> 02:40:16,199
YOU HAVE A 50 PERCENT CHANCE OF 

3258
02:40:16,199 --> 02:40:19,069
GETTING THE RIGHT ANSWER.

3259
02:40:19,069 --> 02:40:21,672
THE BOTTOM ONE, OKAY, YES.

3260
02:40:21,672 --> 02:40:31,248
GIVE HER THE AWARD.

3261
02:40:31,248 --> 02:40:33,750
BECAUSE IT'S HOW THEY WERE MAD.

3262
02:40:33,750 --> 02:40:40,390
SO PCEB WAS MADE FROM MAKING A 

3263
02:40:40,390 --> 02:40:41,758
DELETION.

3264
02:40:41,758 --> 02:40:47,230
IT'S THE MSC SITE TO PST SITE, 

3265
02:40:47,230 --> 02:40:50,033
AND IF YOU'RE A REAL EXPERT, 

3266
02:40:50,033 --> 02:40:52,669
YOU'LL SAY THERE ARE TWO PSII 

3267
02:40:52,669 --> 02:40:55,038
SITES, WHICH ONE?

3268
02:40:55,038 --> 02:40:57,140
WHEREAS CGP WAS MADE BY MOVING A

3269
02:40:57,140 --> 02:40:59,609
FRAGMENT TO EXPRESS IT AT THE 

3270
02:40:59,609 --> 02:41:00,911
CMV PROMOTER.

3271
02:41:00,911 --> 02:41:02,546
AND JUST FORTUITOUSLY, THE SITE 

3272
02:41:02,546 --> 02:41:04,715
THAT THEY PICKED WAS JUST A 

3273
02:41:04,715 --> 02:41:06,383
STREAM OF WHERE GLYCOGAG STARTS.

3274
02:41:06,383 --> 02:41:08,652
JUST TO BRING YOU BACK TO WHAT 

3275
02:41:08,652 --> 02:41:11,188
RESEARCH WAS LIKE IN THE EARLY 

3276
02:41:11,188 --> 02:41:21,698
'80s,  WHEN I JOINED STEVE'S 

3277
02:41:24,301 --> 02:41:25,302
LAB, THIS WAS THE SEQUENCE THAT 

3278
02:41:25,302 --> 02:41:26,303
HE GAVE US.

3279
02:41:26,303 --> 02:41:29,139
IF YOU NOTE THAT GAG, THERE'S A 

3280
02:41:29,139 --> 02:41:31,041
FAINT ERROR, SO IF YOU REALLY 

3281
02:41:31,041 --> 02:41:32,209
WANTED TO KNOW THE SEQUENCE OF 

3282
02:41:32,209 --> 02:41:34,044
GAG, YOU HAD TO COUNT THAT IN 

3283
02:41:34,044 --> 02:41:35,278
THREES, AND YOU CAN SEE VERY 

3284
02:41:35,278 --> 02:41:37,948
HIDDEN THERE PENCILED IN, IT 

3285
02:41:37,948 --> 02:41:40,751
SAYS WHERE GLYCOGAG STARTS, AND 

3286
02:41:40,751 --> 02:41:45,789
YOU COULD SEE HERE THAT BAL 1, 

3287
02:41:45,789 --> 02:41:50,627
WHICH IS NOW MSC 1 TO PST, IF 

3288
02:41:50,627 --> 02:41:52,362
YOU DID THAT INCISION, WRITTEN 

3289
02:41:52,362 --> 02:41:54,064
IN MY HANDWRITING ON THE LEFT, 

3290
02:41:54,064 --> 02:41:56,066
YOU CUT OUT GLYCOGAG.

3291
02:41:56,066 --> 02:42:01,505
BUT IF YOU STARTED THAT XMA 1 

3292
02:42:01,505 --> 02:42:03,840
SITE, YOU WOULD JUST LIKELY HAVE

3293
02:42:03,840 --> 02:42:05,542
GLYCOGAG INCLUDED IN IT.

3294
02:42:05,542 --> 02:42:06,443
SO IF YOU WERE INTERESTED IN 

3295
02:42:06,443 --> 02:42:07,811
WORKING WITH A VECTOR, YOU 

3296
02:42:07,811 --> 02:42:10,414
SHOULD BE WORKING IN ONE THAT 

3297
02:42:10,414 --> 02:42:12,983
STUDIES PARTICLE ENTRY, IT'S THE

3298
02:42:12,983 --> 02:42:13,950
BOTTOM ONE TO USE.

3299
02:42:13,950 --> 02:42:15,018
OF COURSE I DIDN'T REMEMBER 

3300
02:42:15,018 --> 02:42:17,354
THAT, AND WE'RE STILL WORKING 

3301
02:42:17,354 --> 02:42:24,294
WITH PCEB, AND I WENT INTO A 

3302
02:42:24,294 --> 02:42:26,763
PANIC AGAIN.

3303
02:42:26,763 --> 02:42:28,131
ALAN FREQUENTLY GETS ASKED TO 

3304
02:42:28,131 --> 02:42:29,499
WRITE STORIES ABOUT SCIENTISTS 

3305
02:42:29,499 --> 02:42:33,170
WHO PASSED AWAY, AND HE DID 

3306
02:42:33,170 --> 02:42:38,275
WRITE THIS ONE ABOUT STEPHEN 

3307
02:42:38,275 --> 02:42:41,278
ROROSZLAN, AND HE SENT THIS TO 

3308
02:42:41,278 --> 02:42:44,714
PEOPLE WHO HAD STORIES ABOUT 

3309
02:42:44,714 --> 02:42:47,617
STEPHEN OROSZLAN, AND I WROTE 

3310
02:42:47,617 --> 02:42:50,387
THIS STORY ABOUT, WHEN WE FIRST 

3311
02:42:50,387 --> 02:42:53,356
STARTED IN STEPHEN GOFF'S LAB, 

3312
02:42:53,356 --> 02:42:54,791
WE WEREN'T VERY ADVANCED ABOUT 

3313
02:42:54,791 --> 02:42:55,892
WHERE PROTEINS STARTED AND 

3314
02:42:55,892 --> 02:42:57,694
ENDED.

3315
02:42:57,694 --> 02:43:00,030
SO WE HAD NO IDEA WHERE 

3316
02:43:00,030 --> 02:43:02,566
PROTEINED STARTED AND ENDED, SO 

3317
02:43:02,566 --> 02:43:06,336
WE PICKED A SEQUENCE AND PUT IT 

3318
02:43:06,336 --> 02:43:08,338
ON THE END AND TRIED TO PICK THE

3319
02:43:08,338 --> 02:43:10,707
MOST STABLE PROTEIN, FIGURING 

3320
02:43:10,707 --> 02:43:12,275
THE MOST STABLE SOLUBLE PROTEIN 

3321
02:43:12,275 --> 02:43:13,477
IS PROBABLY THE RIGHT THING, AND

3322
02:43:13,477 --> 02:43:15,345
THAT'S WHAT WE WORKED WITH.

3323
02:43:15,345 --> 02:43:17,013
THEN I WENT TO COLD SPRING 

3324
02:43:17,013 --> 02:43:19,382
HARBOR FOR THE FIRST TIME, AND 

3325
02:43:19,382 --> 02:43:22,586
STEPHEN OROSZLAN WAS THERE, 

3326
02:43:22,586 --> 02:43:27,190
DESCRIBING, I THINK, THE 

3327
02:43:27,190 --> 02:43:29,593
TERMINII OF RT, AND I 

3328
02:43:29,593 --> 02:43:31,428
IMMEDIATELY HAD TO FIND HIM.

3329
02:43:31,428 --> 02:43:32,929
I FOUND HIM IN THE HALLWAY IN 

3330
02:43:32,929 --> 02:43:34,231
FRONT OF THE BAR, WE ALL KNOW 

3331
02:43:34,231 --> 02:43:35,999
WHERE THAT IS, AND I CORNERED 

3332
02:43:35,999 --> 02:43:37,701
HIM TO FIND OUT WHERE THE REAL 

3333
02:43:37,701 --> 02:43:40,437
SEQUENCE WAS, AND HE TOLD ME 

3334
02:43:40,437 --> 02:43:41,605
THERE WOULD BE A MANUSCRIPT 

3335
02:43:41,605 --> 02:43:43,573
COMING OUT VERY SOON.

3336
02:43:43,573 --> 02:43:45,342
IT WAS ACTUALLY RELEASED RIGHT 

3337
02:43:45,342 --> 02:43:47,777
AFTER THE MEETING, SO HE 

3338
02:43:47,777 --> 02:43:48,879
ACTUALLY DID KNOW THE ANSWER, 

3339
02:43:48,879 --> 02:43:51,114
BUT HI TO WAIT FOR THE PAPER TO 

3340
02:43:51,114 --> 02:43:52,249
COME OUT.

3341
02:43:52,249 --> 02:43:54,084
IT TURNS OUT WE WERE PRETTY 

3342
02:43:54,084 --> 02:44:00,190
CLOSE TO THE REAL AUTHENTIC 

3343
02:44:00,190 --> 02:44:00,423
TERMINI.

3344
02:44:00,423 --> 02:44:03,560
I WROTE THAT STORY TO ALAN, AND 

3345
02:44:03,560 --> 02:44:05,462
I COULD HEAR HIS VOICE SAYING, 

3346
02:44:05,462 --> 02:44:06,763
MONICA, THANKS.

3347
02:44:06,763 --> 02:44:08,265
THAT'S A VERY NICE LITTLE 

3348
02:44:08,265 --> 02:44:08,565
ANECDOTE.

3349
02:44:08,565 --> 02:44:10,267
THE REST OF IT BASICALLY SAYING 

3350
02:44:10,267 --> 02:44:12,903
IT'S PROBABLY NOT GOING TO BE 

3351
02:44:12,903 --> 02:44:15,872
INCORPORATED INTO IT.

3352
02:44:15,872 --> 02:44:16,239
(LAUGHTER) 

3353
02:44:16,239 --> 02:44:18,008
BUT THE BOTTOM LINE IS ALSO A 

3354
02:44:18,008 --> 02:44:19,543
CLASSIC ALAN.

3355
02:44:19,543 --> 02:44:22,712
THANKS AGAIN AND BEST REGARDS.

3356
02:44:22,712 --> 02:44:25,282
SO THIS WAS THE SUMMER OF THE 

3357
02:44:25,282 --> 02:44:28,685
THING THAT ALAN WROTE ABOUT 

3358
02:44:28,685 --> 02:44:31,388
STEVE OROSZLAN, AND THREE 

3359
02:44:31,388 --> 02:44:33,490
QUARTERS OF IT IS ABOUT HIV, BUT

3360
02:44:33,490 --> 02:44:35,325
THE VERY LAST PARAGRAPH IS ABOUT

3361
02:44:35,325 --> 02:44:38,261
DESCRIBING WHEN STEVE OROSZLAN 

3362
02:44:38,261 --> 02:44:41,665
FOUND A SEQUENCE OF P12, AND IT 

3363
02:44:41,665 --> 02:44:46,570
HAD A PPPY SFENS IN IT, AND 

3364
02:44:46,570 --> 02:44:53,410
STEVE SAID HE ELICITED A LOUD 

3365
02:44:53,410 --> 02:44:56,012
SIGH OF AMAZEMENT AND INCLUDING 

3366
02:44:56,012 --> 02:45:00,116
AN EXPLETIVE, TURNS OUT THAT'S 

3367
02:45:00,116 --> 02:45:04,387
THE LAST DOMAIN AND P12.

3368
02:45:04,387 --> 02:45:07,157
SO I WAS AT LEAST HAPPY THAT P12

3369
02:45:07,157 --> 02:45:07,791
WAS INCORPORATED INTO THAT 

3370
02:45:07,791 --> 02:45:09,926
STORY.

3371
02:45:09,926 --> 02:45:13,129
THEN I PUBLISHED OUR WORK ON 

3372
02:45:13,129 --> 02:45:15,565
P12, IN WHICH WE TOOK OUT THE 

3373
02:45:15,565 --> 02:45:16,900
REGION WE THOUGHT WAS THE 

3374
02:45:16,900 --> 02:45:18,802
TETHERING DOMAIN AND INSERTED IN

3375
02:45:18,802 --> 02:45:20,337
TETHERING DOMAINS FROM LOTS OF 

3376
02:45:20,337 --> 02:45:21,404
DIFFERENT VIRUSES AND FOUND OUT 

3377
02:45:21,404 --> 02:45:23,139
THAT THEY SUBSTITUTED.

3378
02:45:23,139 --> 02:45:26,209
BUT MORE TOUCHING WAS THAT ALAN 

3379
02:45:26,209 --> 02:45:28,078
WROTE THE COMMENTARY ABOUT THIS 

3380
02:45:28,078 --> 02:45:30,413
STUDY, AND IT REALLY WAS 

3381
02:45:30,413 --> 02:45:33,049
SIGNIFICANT TO ME THAT ALAN 

3382
02:45:33,049 --> 02:45:35,085
WROTE THIS COMMENTARY ABOUT OUR 

3383
02:45:35,085 --> 02:45:40,924
WORK, SO THANK YOU.

3384
02:45:40,924 --> 02:45:43,460
I HAVE SOME MORE POLITE E-MAILS 

3385
02:45:43,460 --> 02:45:44,661
FROM ALAN.

3386
02:45:44,661 --> 02:45:48,598
USUALLY WHEN HE'S LOOKING FOR A 

3387
02:45:48,598 --> 02:45:49,933
REAGENT, AND YOU CAN TRY TO 

3388
02:45:49,933 --> 02:45:52,135
GUESS WHAT HE'S WORKING ON 

3389
02:45:52,135 --> 02:45:53,870
BECAUSE I HAD NO IDEA WHY I WAS 

3390
02:45:53,870 --> 02:45:55,138
ASKING FOR THESE REAGENTS.

3391
02:45:55,138 --> 02:45:57,741
THIS IS ONE HE WROTE TO LORRAINE

3392
02:45:57,741 --> 02:46:00,543
AND MYSELF, LOOKING FOR 

3393
02:46:00,543 --> 02:46:08,885
REPLICATION COMPETENT MLV WITH A

3394
02:46:08,885 --> 02:46:09,986
XENOTROPIC EMV.

3395
02:46:09,986 --> 02:46:12,188
AND I DON'T HAVE IT.

3396
02:46:12,188 --> 02:46:14,891
HERE'S A CLASSIC ALAN ANSWER, 

3397
02:46:14,891 --> 02:46:16,259
OKAY, BEST REGARDS ANYWAY.

3398
02:46:16,259 --> 02:46:17,527
HERE'S ANOTHER ONE FROM OCTOBER 

3399
02:46:17,527 --> 02:46:20,397
9th, 2020.

3400
02:46:20,397 --> 02:46:22,932
LOOKING, ASKING ABOUT P50.

3401
02:46:22,932 --> 02:46:29,472
FOR THOSE OF YOU WHO WORKED IN 

3402
02:46:29,472 --> 02:46:31,508
MLV, THERE'S THIS P50.

3403
02:46:31,508 --> 02:46:35,679
IT HAS MA, P12, CA, AND SPLICES 

3404
02:46:35,679 --> 02:46:39,149
INTO THE C TERMINUS OF IN.

3405
02:46:39,149 --> 02:46:44,421
SO HE ASKED ME IF I HAD ANY 

3406
02:46:44,421 --> 02:46:46,656
ANTISERA TO TRY, WHICH IS 

3407
02:46:46,656 --> 02:46:47,891
ANTI-MA OR ANTI-IN.

3408
02:46:47,891 --> 02:46:50,794
IF YOU LOOK AT THE DATE, IT'S 

3409
02:46:50,794 --> 02:46:52,796
OCTOBER 9th, 2020, WHEN BOTH OF 

3410
02:46:52,796 --> 02:46:54,731
US WERE AT HOME BEING LOCKED OUT

3411
02:46:54,731 --> 02:46:56,299
OF OUR LABS DURING COVID.

3412
02:46:56,299 --> 02:46:58,868
I SAID, ALAN, IT'S GOING TO TAKE

3413
02:46:58,868 --> 02:47:01,371
A LITTLE DIGGING IF I STORE IT 

3414
02:47:01,371 --> 02:47:01,838
ANYWHERE.

3415
02:47:01,838 --> 02:47:03,740
I'VE BEEN WORKING AT HOME SINCE 

3416
02:47:03,740 --> 02:47:04,808
MARCH.

3417
02:47:04,808 --> 02:47:05,809
I LITERALLY WAS GIVEN TWO HOURS 

3418
02:47:05,809 --> 02:47:07,677
TO LEAVE THE LAB AND NEVER COME 

3419
02:47:07,677 --> 02:47:09,813
BACK FOR A YEAR.

3420
02:47:09,813 --> 02:47:13,616
I ACTUALLY DIDN'T TAKE MY OWN 

3421
02:47:13,616 --> 02:47:20,724
IPs WITH MLV SERA.

3422
02:47:20,724 --> 02:47:22,358
ANYWAY, THAT'S HOW WE TRIED TO 

3423
02:47:22,358 --> 02:47:23,426
DO SCIENCE DURING COVID.

3424
02:47:23,426 --> 02:47:24,961
HE SAID, I KNOW WHAT YOU MEAN.

3425
02:47:24,961 --> 02:47:27,430
I'M HOME SEPARATED FROM MY 

3426
02:47:27,430 --> 02:47:27,630
FILES.

3427
02:47:27,630 --> 02:47:29,299
I NEVER DID -- I APOLOGIZE.

3428
02:47:29,299 --> 02:47:31,067
I NEVER DID GO BACK TO LOOK FOR 

3429
02:47:31,067 --> 02:47:33,703
THOSE, BUT I DO WANT TO HAVE A 

3430
02:47:33,703 --> 02:47:34,671
COMMERCIAL HERE SAYING, IF 

3431
02:47:34,671 --> 02:47:38,041
YOU'RE LOOKING FOR MLV 

3432
02:47:38,041 --> 02:47:44,681
ANTIBODIES, CHRISTINE KOZACK HAS

3433
02:47:44,681 --> 02:47:47,050
FULL FROM THE ONES FROM THE 

3434
02:47:47,050 --> 02:47:47,350
REPOSITORY.

3435
02:47:47,350 --> 02:47:48,852
IF YOU ARE LOOKING FOR THOSE, 

3436
02:47:48,852 --> 02:47:51,387
THEY'RE GREAT ANTIBODIES.

3437
02:47:51,387 --> 02:48:00,263
THEY'RE PROBABLY NICE VIALS, AND

3438
02:48:00,263 --> 02:48:01,731
WE'D HATE TO HAVE THEM THROW 

3439
02:48:01,731 --> 02:48:01,931
OUT.

3440
02:48:01,931 --> 02:48:03,900
IF PEOPLE KNOW WHAT TO DO WITH 

3441
02:48:03,900 --> 02:48:06,669
THIS SERUM, WE'D CERTAINLY LIKE 

3442
02:48:06,669 --> 02:48:08,671
THEM TO HAVE A HOME BEFORE 

3443
02:48:08,671 --> 02:48:11,074
CHRISTINE RETIRES OR ANYONE 

3444
02:48:11,074 --> 02:48:11,908
ELSE.

3445
02:48:11,908 --> 02:48:13,042
I DO HAVE AN APOLOGY.

3446
02:48:13,042 --> 02:48:15,612
I'M SURPRISED THAT ALAN ASKED ME

3447
02:48:15,612 --> 02:48:17,514
FOR REAGENTS BECAUSE ACTUALLY IN

3448
02:48:17,514 --> 02:48:20,150
2015 I HAD SENT HIM SOME SAMPLES

3449
02:48:20,150 --> 02:48:23,353
OF SOME P12 MUTANTS, TRYING TO 

3450
02:48:23,353 --> 02:48:27,257
LOOK AT THE SHAPE OF THE CAPCID.

3451
02:48:27,257 --> 02:48:28,925
UNFORTUNATELY, THIS IS WHAT I 

3452
02:48:28,925 --> 02:48:30,126
GOT BACK WHERE THEY SAID VERY 

3453
02:48:30,126 --> 02:48:32,328
LAST SIX IMAGES DISPLAYED, 

3454
02:48:32,328 --> 02:48:35,832
POSSIBLE MICROPLASM OR YEAST 

3455
02:48:35,832 --> 02:48:36,666
CONTAMINATION.

3456
02:48:36,666 --> 02:48:38,468
SO I AM HORRIFIED STILL TO THIS 

3457
02:48:38,468 --> 02:48:40,637
DAY THAT I SENT SAMPLES THAT 

3458
02:48:40,637 --> 02:48:42,438
MIGHT NOT HAVE BEEN AS CLEAN AS 

3459
02:48:42,438 --> 02:48:44,007
THEY EVER WERE.

3460
02:48:44,007 --> 02:48:45,642
LITERALLY, MY LAB HAS NEVER 

3461
02:48:45,642 --> 02:48:50,880
TAKEN ANY CULTURE AT A PLASMA 

3462
02:48:50,880 --> 02:48:52,182
SCENE SINCE THEN BECAUSE WE 

3463
02:48:52,182 --> 02:48:53,650
DON'T EVER WANT THAT TO HAPPEN.

3464
02:48:53,650 --> 02:48:57,153
MY APOLOGIES IF WE CONTAMINATED 

3465
02:48:57,153 --> 02:48:57,587
ANYTHING.

3466
02:48:57,587 --> 02:48:59,656
I'M STILL SHAKING FROM THAT.

3467
02:48:59,656 --> 02:49:01,658
HERE'S ANOTHER E-MAIL THAT WAS 

3468
02:49:01,658 --> 02:49:06,329
JUST LAST YEAR.

3469
02:49:06,329 --> 02:49:07,997
WE WERE AT THE 40th ANNIVERSARY 

3470
02:49:07,997 --> 02:49:14,771
OF HIV, AND I WAS SPEAKING WITH 

3471
02:49:14,771 --> 02:49:24,514
AT ALLEGE ARE  ALICE AND WES.

3472
02:49:24,514 --> 02:49:27,116
THE SOLUTION WAS LET'S ASK ALAN 

3473
02:49:27,116 --> 02:49:27,450
REiNS.

3474
02:49:27,450 --> 02:49:29,919
THIS WAS AT 4:54 COFFEE BREAK.

3475
02:49:29,919 --> 02:49:31,654
I SENT HIM AN E-MAIL.

3476
02:49:31,654 --> 02:49:33,022
LITERALLY FOUR MINUTES LATER WE 

3477
02:49:33,022 --> 02:49:33,990
GOT THE ANSWER.

3478
02:49:33,990 --> 02:49:35,258
THE ANSWER TO THE QUESTION IS 

3479
02:49:35,258 --> 02:49:36,793
YES, AND I'M HOPING TO PUBLISH 

3480
02:49:36,793 --> 02:49:39,596
THE PAPER BEFORE I DIE OF OLD 

3481
02:49:39,596 --> 02:49:42,599
AGE, GLAD SOMEONE IS INTERESTED.

3482
02:49:42,599 --> 02:49:45,802
SO I'M HAPPY TO SAY THAT THIS 

3483
02:49:45,802 --> 02:49:51,107
PAPER DID COME OUT IN MBIO IN 

3484
02:49:51,107 --> 02:49:51,474
JULY.

3485
02:49:51,474 --> 02:50:01,951
SO, YES, THERE'S THE ANSWER.

3486
02:50:06,022 --> 02:50:07,323
>> HE'S WITH US.

3487
02:50:07,323 --> 02:50:07,757
>> RAISE YOUR HAND.

3488
02:50:07,757 --> 02:50:10,660
THANK YOU.

3489
02:50:10,660 --> 02:50:12,028
THIS IS A SLIGHT DEVIATION FROM 

3490
02:50:12,028 --> 02:50:12,929
THE TOPIC.

3491
02:50:12,929 --> 02:50:16,666
I WENT TO MY SCHOOL'S TRYING TO 

3492
02:50:16,666 --> 02:50:18,234
DISCUSS OUR NEW ENTERING CLASS 

3493
02:50:18,234 --> 02:50:20,303
OF MEDICAL STUDENTS, AND THIS 

3494
02:50:20,303 --> 02:50:23,306
WAS ONE OF THEIR SLIDES ABOUT A 

3495
02:50:23,306 --> 02:50:27,610
SNAPSHOT OF THE PEOPLE GOING OR 

3496
02:50:27,610 --> 02:50:30,246
ENTERING ROBERT W. JOHNSON 

3497
02:50:30,246 --> 02:50:32,148
MEDICAL SCHOOL CLASS, AND THE 

3498
02:50:32,148 --> 02:50:33,349
QUESTION WAS HOW MANY PEOPLE 

3499
02:50:33,349 --> 02:50:35,184
SPEAK DIFFERENT LANGUAGES AT 

3500
02:50:35,184 --> 02:50:35,652
HOME.

3501
02:50:35,652 --> 02:50:36,853
THEY SAID 40 PERCENT OF THE 

3502
02:50:36,853 --> 02:50:38,021
CLASS SPEAKS ONE OR TWO 

3503
02:50:38,021 --> 02:50:39,322
DIFFERENT LANGUAGES AT A NATIVE 

3504
02:50:39,322 --> 02:50:40,957
OR ADVANCED LEVEL AT HOME.

3505
02:50:40,957 --> 02:50:43,226
IT'S REALLY AN IMPRESSIVE STORY,

3506
02:50:43,226 --> 02:50:45,828
AND IT BASICALLY SHOWS YOU WHO'S

3507
02:50:45,828 --> 02:50:47,764
GOING IN TO BE CLINICIANS THESE 

3508
02:50:47,764 --> 02:50:47,964
DAYS.

3509
02:50:47,964 --> 02:50:51,200
SO WHEN I SAW THIS, I'M SURE 

3510
02:50:51,200 --> 02:50:52,769
EVERYONE ELSE HERE IS THINKING, 

3511
02:50:52,769 --> 02:50:54,137
OH, WHAT LANGUAGES WERE SPOKEN 

3512
02:50:54,137 --> 02:50:55,672
IN MY HOUSE?

3513
02:50:55,672 --> 02:50:58,207
BOTH OF MY PARENTS WERE BORN IN 

3514
02:50:58,207 --> 02:50:59,676
BROOKLYN, WHICH SOME PEOPLE 

3515
02:50:59,676 --> 02:51:01,644
MIGHT THINK IS A FOREIGN 

3516
02:51:01,644 --> 02:51:05,682
LANGU

3517
02:51:05,682 --> 02:51:06,816
LANGUAGE, AND IF YOU REALLY WANT

3518
02:51:06,816 --> 02:51:08,151
TO TALK ABOUT IT, WE CAN HAVE IT

3519
02:51:08,151 --> 02:51:14,257
OVER COFFEE.

3520
02:51:14,257 --> 02:51:15,992
DESPITE BEING BORN IN BROOKLYN, 

3521
02:51:15,992 --> 02:51:19,829
MY MOM WAS A YIDDISH-IST, AND 

3522
02:51:19,829 --> 02:51:22,231
SHE TRIED HER HARDEST TO KEEP 

3523
02:51:22,231 --> 02:51:24,467
THIS LANGUAGE FROM DYING, SO SHE

3524
02:51:24,467 --> 02:51:27,103
SENT ME TO YIDDISH SCHOOL FOR 12

3525
02:51:27,103 --> 02:51:28,938
YEARS, MAYBE 15 YEARS.

3526
02:51:28,938 --> 02:51:31,641
WHEN ALAN HEARD THIS, HE STARTED

3527
02:51:31,641 --> 02:51:34,510
SENDING ME NEWSLETTERS FROM KLEZ

3528
02:51:34,510 --> 02:51:36,946
CALIFORNIA, AND I THOUGHT I KNEW

3529
02:51:36,946 --> 02:51:38,314
EVERY YIDDISH ORGANIZATION EVER 

3530
02:51:38,314 --> 02:51:40,216
BECAUSE OF MY MOM, BUT I 

3531
02:51:40,216 --> 02:51:43,052
ACTUALLY HAD NEVER HEARD OF KLEZ

3532
02:51:43,052 --> 02:51:43,353
CALIFORNIA.

3533
02:51:43,353 --> 02:51:44,887
HE SENT ME ONE, AND I WAS 

3534
02:51:44,887 --> 02:51:45,722
LOOKING THROUGH IT.

3535
02:51:45,722 --> 02:51:48,191
SOME OF YOU MIGHT HAVE HEARD OF 

3536
02:51:48,191 --> 02:51:51,094
PETE SEGER.

3537
02:51:51,094 --> 02:51:53,062
THE OTHER PERSON IN THE PICTURE 

3538
02:51:53,062 --> 02:51:56,566
IS RUTH RUBIN, WHO IS A VERY 

3539
02:51:56,566 --> 02:51:59,135
FAMOUS FOLKLORIST IN YIDDISH 

3540
02:51:59,135 --> 02:52:00,436
MUSIC, AND MY MOM KNEW HER AND 

3541
02:52:00,436 --> 02:52:01,571
WORKED WITH HER DIRECTLY.

3542
02:52:01,571 --> 02:52:03,339
SHE REALLY IS ONE OF THESE 

3543
02:52:03,339 --> 02:52:04,140
FAMOUS PEOPLE.

3544
02:52:04,140 --> 02:52:05,308
WE HAD HER BOOKS AT HOME.

3545
02:52:05,308 --> 02:52:07,777
WHEN I SAW THIS, I WENT AND I 

3546
02:52:07,777 --> 02:52:10,480
WATCHED THIS LINK, AND THEY WERE

3547
02:52:10,480 --> 02:52:14,917
SINGING THIS SONG.

3548
02:52:14,917 --> 02:52:16,953
JUST LIKE LOOKING BACK, WHY WAS 

3549
02:52:16,953 --> 02:52:17,987
I DOING THIS?

3550
02:52:17,987 --> 02:52:19,455
WHAT WAS MY FRAMEWORK THAT I 

3551
02:52:19,455 --> 02:52:21,224
STARTED TO GO BACK AND LOOK AT 

3552
02:52:21,224 --> 02:52:23,493
YIDDISH SONGS FROM THE EARLY 

3553
02:52:23,493 --> 02:52:30,633
1900s?

3554
02:52:30,633 --> 02:52:31,734
I LOOKED AT THE DATE OF THE 

3555
02:52:31,734 --> 02:52:34,637
E-MAIL I SENT TO ALAN, NOVEMBER 

3556
02:52:34,637 --> 02:52:43,613
7, 2022.

3557
02:52:43,613 --> 02:52:47,784
IT SAYS, WITH THE ELECTIONS 

3558
02:52:47,784 --> 02:52:51,587
TOMORROW, IT'S HARD TO WATCH TV,

3559
02:52:51,587 --> 02:52:54,323
AND I'VE BEEN RESORTING TO EASY 

3560
02:52:54,323 --> 02:52:57,193
WATCHING LIKE BRITISH BAKING 

3561
02:52:57,193 --> 02:52:58,327
SHOW TWO.

3562
02:52:58,327 --> 02:53:01,631
>> BAKEOFF, ISN'T IT?

3563
02:53:01,631 --> 02:53:04,567
>> IN ENGLAND, IT'S CALLED
THE 

3564
02:53:04,567 --> 02:53:06,002
BAKEOFF, BUT IT'S THE BRITISH 

3565
02:53:06,002 --> 02:53:06,903
BAKE SHOW TWO.

3566
02:53:06,903 --> 02:53:08,838
AND IT JUST REALLY HIT ME 

3567
02:53:08,838 --> 02:53:10,540
BECAUSE HERE I AM TWO YEARS 

3568
02:53:10,540 --> 02:53:12,375
LATER, I CAN'T WATCH THE NEWS.

3569
02:53:12,375 --> 02:53:14,444
I'VE BEEN WATCHING THE BRITISH 

3570
02:53:14,444 --> 02:53:17,447
BAKE SHOW TWO, AND THERE'S A NEW

3571
02:53:17,447 --> 02:53:20,917
EPISODE TONIGHT.

3572
02:53:20,917 --> 02:53:24,353
SO IF YOU ALSO CAN'T WATCH THE 

3573
02:53:24,353 --> 02:53:27,557
NEWS, I'D HIGHLY RECOMMEND 

3574
02:53:27,557 --> 02:53:29,759
WATCHING THE SHOW.

3575
02:53:29,759 --> 02:53:31,928
UNLIKE AMERICAN TV, WHERE PEOPLE

3576
02:53:31,928 --> 02:53:33,362
COMPETE WITH EACH OTHER AND BACK

3577
02:53:33,362 --> 02:53:34,163
STAB EACH OTHER.

3578
02:53:34,163 --> 02:53:35,932
IN THIS SHOW, THEY ACTUALLY DO 

3579
02:53:35,932 --> 02:53:37,934
COMPETE EACH OTHER, BUT THEY 

3580
02:53:37,934 --> 02:53:39,001
ACTUALLY HELP EACH OTHER AND 

3581
02:53:39,001 --> 02:53:40,069
THEY LIKE EACH OTHER.

3582
02:53:40,069 --> 02:53:42,105
I REALLY THINK THAT'S WHERE WE 

3583
02:53:42,105 --> 02:53:44,474
SHOULD BE GOING.

3584
02:53:44,474 --> 02:53:46,642
I ALSO HEAR PUT A MOVIE 

3585
02:53:46,642 --> 02:53:48,411
RECOMMENDATION THAT I WATCHED, 

3586
02:53:48,411 --> 02:53:50,346
WHICH IS THE PATIENT, WHICH ALSO

3587
02:53:50,346 --> 02:53:51,147
WE COMMUNICATE WHERE YOU CAN 

3588
02:53:51,147 --> 02:53:53,182
STREAM IT, SO IT WAS ON HULU.

3589
02:53:53,182 --> 02:53:54,750
IF YOU'VE EVER WATCHED THAT, IT 

3590
02:53:54,750 --> 02:53:56,352
WAS PRETTY INTENSE IN OTHER 

3591
02:53:56,352 --> 02:53:56,752
WAYS.

3592
02:53:56,752 --> 02:53:59,155
SO I WAS LOOKING FORWARD TO SOME

3593
02:53:59,155 --> 02:54:03,159
MORE SUGGESTIONS, AND HERE I AM.

3594
02:54:03,159 --> 02:54:05,761
SO WHAT ABOUT THIS SONG, WHY DID

3595
02:54:05,761 --> 02:54:06,729
I KNOW THIS SONG?

3596
02:54:06,729 --> 02:54:10,066
THIS IS THE TRANSLATION OF IT.

3597
02:54:10,066 --> 02:54:12,435
IT TALKS ABOUT THESE TWO TOWNS 

3598
02:54:12,435 --> 02:54:18,241
THAT HAVE VERY FUNNY NAMES, 

3599
02:54:18,241 --> 02:54:26,149
SVASTOPOL AND SIFARUPPUL AND 

3600
02:54:26,149 --> 02:54:31,320
THIS TOWN NEARBY CALLED 

3601
02:54:31,320 --> 02:54:35,124
ZHANKOYE, AND IT'S A FARM WHERE 

3602
02:54:35,124 --> 02:54:37,426
GRANDPA DRIVES THE TRACTOR AND 

3603
02:54:37,426 --> 02:54:38,828
GRANDMA RUNS THE CREAM 

3604
02:54:38,828 --> 02:54:40,029
EXTRACTOR.

3605
02:54:40,029 --> 02:54:43,065
I SAID, WHERE IS THIS IDYLLIC 

3606
02:54:43,065 --> 02:54:44,934
PLACE, AND YOU LOOK AT THE MAP, 

3607
02:54:44,934 --> 02:54:47,670
AND WE'RE RIGHT IN THE MIDDLE OF

3608
02:54:47,670 --> 02:54:50,006
CRIMEA AND UKRAINE, NOT QUITE 

3609
02:54:50,006 --> 02:54:56,979
THESE DAYS AN IDYLLIC PLACE.

3610
02:54:56,979 --> 02:55:01,350
SO I WANT TO THANK ALAN FOR 

3611
02:55:01,350 --> 02:55:03,052
GOING DOWN THIS TRIP TO MEMORY 

3612
02:55:03,052 --> 02:55:03,252
LANE.

3613
02:55:03,252 --> 02:55:04,754
I HEARD A STATEMENT, WHEN IT 

3614
02:55:04,754 --> 02:55:06,589
COMES TO TIME TRAVEL, SCIENTISTS

3615
02:55:06,589 --> 02:55:08,758
ALWAYS CHOOSE THE FUTURE BECAUSE

3616
02:55:08,758 --> 02:55:17,733
WE CANNOT GO BACK.

3617
02:55:17,733 --> 02:55:20,536
WE CANNOT GO BACK.

3618
02:55:20,536 --> 02:55:23,506
SO NOW WE'LL GET INTO SOME OTHER

3619
02:55:23,506 --> 02:55:26,676
MOV

3620
02:55:26,676 --> 02:55:27,543
MOVIES.

3621
02:55:27,543 --> 02:55:29,378
ACTUALLY COMING HERE, I WAS 

3622
02:55:29,378 --> 02:55:31,847
TRYING TO LOOK UP MY TSA NUMBER 

3623
02:55:31,847 --> 02:55:33,649
BECAUSE SOMEHOW IT WASN'T ON MY 

3624
02:55:33,649 --> 02:55:35,551
TICKET AND I WASN'T GOING TO 

3625
02:55:35,551 --> 02:55:38,187
TAKE MY SHOES OFF AND TAKE MY 

3626
02:55:38,187 --> 02:55:39,522
COMPUTER OUT.

3627
02:55:39,522 --> 02:55:40,556
I WAS SEARCHING THROUGH OLD 

3628
02:55:40,556 --> 02:55:42,892
NOTES THAT I HAD ON MY PHONE, 

3629
02:55:42,892 --> 02:55:45,261
AND I ACTUALLY FOUND THE NOTE 

3630
02:55:45,261 --> 02:55:49,699
FROM 2016 THAT SAYS REIN EX 

3631
02:55:49,699 --> 02:55:49,999
MACHINA.

3632
02:55:49,999 --> 02:55:51,601
THAT WAS ONE OF THE FIRST MOVIES

3633
02:55:51,601 --> 02:55:53,603
THAT HE HAD SUGGESTED THAT I 

3634
02:55:53,603 --> 02:55:53,803
WATCH.

3635
02:55:53,803 --> 02:55:56,572
I DON'T KNOW HOW MANY OF YOU 

3636
02:55:56,572 --> 02:55:58,140
WATCHED IT.

3637
02:55:58,140 --> 02:55:58,641
NO?

3638
02:55:58,641 --> 02:56:02,211
ANYWAY, IT'S A GREAT MOVIE.

3639
02:56:02,211 --> 02:56:06,048
IT'S AI BEFORE THERE WAS AI, SO 

3640
02:56:06,048 --> 02:56:11,087
IF YOU THINK OF 2001 AND HAL AS 

3641
02:56:15,258 --> 02:56:17,593
DEPICTING THE FUTURE, WELL, EX 

3642
02:56:17,593 --> 02:56:19,996
MACHINA IS REALLY PREDICTING 

3643
02:56:19,996 --> 02:56:21,697
WHERE WE ARE THESE DAYS.

3644
02:56:21,697 --> 02:56:24,166
THE RUNNING LINE IS TO ERASE THE

3645
02:56:24,166 --> 02:56:25,801
LINE BETWEEN MAN AND MACHINE IS 

3646
02:56:25,801 --> 02:56:28,170
TO OBSCURE THE LINE BETWEEN MEN 

3647
02:56:28,170 --> 02:56:29,138
AND GODS.

3648
02:56:29,138 --> 02:56:30,640
DON'T YOU WANT TO WATCH THAT 

3649
02:56:30,640 --> 02:56:31,807
MOVIE?

3650
02:56:31,807 --> 02:56:32,942
MIRACULOUSLY, IT'S NOW STREAMING

3651
02:56:32,942 --> 02:56:34,176
ON AMAZON.

3652
02:56:34,176 --> 02:56:37,346
IT JUST CAME BACK TO BE ON 

3653
02:56:37,346 --> 02:56:37,613
AMAZON.

3654
02:56:37,613 --> 02:56:39,849
SO IF YOU DON'T WANT TO WATCH 

3655
02:56:39,849 --> 02:56:42,852
THE BRITISH BAKE SHOW, YOU CAN 

3656
02:56:42,852 --> 02:56:50,793
WATCH EX MACHINA AND AFTER THAT 

3657
02:56:50,793 --> 02:56:51,861
RECOMMENDATION FROM ALAN, I 

3658
02:56:51,861 --> 02:56:53,963
TRUST ALL OF HIS 

3659
02:56:53,963 --> 02:56:54,330
RECOMMENDATIONS.

3660
02:56:54,330 --> 02:56:55,531
HERE'S MORE MOVIE SUGGESTIONS 

3661
02:56:55,531 --> 02:57:03,306
FROM ALAN.

3662
02:57:03,306 --> 02:57:07,410
AND SOME OF HIS COMMENTS.

3663
02:57:07,410 --> 02:57:09,879
OUTLANDER IS COMPELLING, NOT TO 

3664
02:57:09,879 --> 02:57:11,847
SAY ADDICTIVE, BUT I THINK HE 

3665
02:57:11,847 --> 02:57:13,949
DOESN'T WATCH IT ANYMORE.

3666
02:57:13,949 --> 02:57:15,618
AND THEN THE TRIAL OF THE 

3667
02:57:15,618 --> 02:57:17,320
CHICAGO SEVEN ON NETFLIX, TOOK 

3668
02:57:17,320 --> 02:57:19,955
ME RIGHT BACK TO 1968.

3669
02:57:19,955 --> 02:57:20,990
AMAZING, MUST WATCH.

3670
02:57:20,990 --> 02:57:22,491
THAT WAS A VERY GOOD MOVIE, I 

3671
02:57:22,491 --> 02:57:23,159
DID WATCH IT.

3672
02:57:23,159 --> 02:57:25,094
THE DANISH GIRL, NOT TO BE 

3673
02:57:25,094 --> 02:57:25,361
MISSED.

3674
02:57:25,361 --> 02:57:26,195
I AGREE.

3675
02:57:26,195 --> 02:57:27,897
THE SIXTH SENSE IS DYNAMITE.

3676
02:57:27,897 --> 02:57:31,367
THEN SOME OTHER ONES THAT I HAD 

3677
02:57:31,367 --> 02:57:33,035
FROM DIFFERENT E-MAILS, 

3678
02:57:33,035 --> 02:57:35,738
OPPENHEIMER, THE 3 BODY PROBLEM,

3679
02:57:35,738 --> 02:57:38,174
AMERICAN FICTION, DISOBEDIENCE, 

3680
02:57:38,174 --> 02:57:42,445
THE QUEENS GAMBIT, AND ALAN 

3681
02:57:42,445 --> 02:57:45,614
WROTE, THE SOPHIE LOREN MOVIE, 

3682
02:57:45,614 --> 02:57:48,384
AND IF YOU ACTUALLY GOOGLE THE 

3683
02:57:48,384 --> 02:57:50,486
SOPHIE LOREN MOVIE, YOU'LL 

3684
02:57:50,486 --> 02:57:51,821
ACTUALLY FIND THE TITLE OF IT.

3685
02:57:51,821 --> 02:57:52,888
IT ACTUALLY DOES WORK.

3686
02:57:52,888 --> 02:57:56,959
SO I WATCHED THEM, AND I AGREE.

3687
02:57:56,959 --> 02:57:58,661
SORT OF IN CONCLUSION, I THINK 

3688
02:57:58,661 --> 02:58:00,529
ALAN IN HIS RESEARCH AND IN HIS 

3689
02:58:00,529 --> 02:58:04,467
MOVIES ARE ALWAYS ON THE CUTTING

3690
02:58:04,467 --> 02:58:06,869
EDGE, AND AS YOU KNOW, THE NOBEL

3691
02:58:06,869 --> 02:58:09,472
PRIZE IN CHEMISTRY THIS YEAR WAS

3692
02:58:09,472 --> 02:58:12,041
ABOUT AI AND THE DEVELOPMENT OF 

3693
02:58:12,041 --> 02:58:15,177
AI MODELS TO STUDY PROTEIN 

3694
02:58:15,177 --> 02:58:15,478
PREDICTIONS.

3695
02:58:15,478 --> 02:58:17,646
I'M NOT TALKING ABOUT MY OWN 

3696
02:58:17,646 --> 02:58:19,915
WORK, BUT WE ACTUALLY DURING 

3697
02:58:19,915 --> 02:58:21,484
COVID, BECAUSE WE COULDN'T DO 

3698
02:58:21,484 --> 02:58:23,252
WET BENCH WORK, WE ACTUALLY 

3699
02:58:23,252 --> 02:58:26,055
STARTED TO DO PROTEIN PREDICTION

3700
02:58:26,055 --> 02:58:27,757
STUDIES, AND WE HAVE A PAPER NOW

3701
02:58:27,757 --> 02:58:30,126
COMING OUT -- WELL, IT WAS 

3702
02:58:30,126 --> 02:58:32,094
SUBMIT

3703
02:58:32,094 --> 02:58:33,262
SUBMITTED.

3704
02:58:33,262 --> 02:58:35,097
STUDYING HOW RECEPTOR 

3705
02:58:35,097 --> 02:58:36,966
TRANSPORTERS WHO HAVE TWO 

3706
02:58:36,966 --> 02:58:40,536
DIFFERENT CONFIRMATIONS.

3707
02:58:40,536 --> 02:58:42,705
IF YOU USE ALPHA FOLD, YOU'LL 

3708
02:58:42,705 --> 02:58:44,640
PROBABLY ONLY GET ONE OF THE TWO

3709
02:58:44,640 --> 02:58:45,641
CONFIRMATIONS, AND WE WORKED OUT

3710
02:58:45,641 --> 02:58:48,577
A WAY, A METHOD TO GET BOTH 

3711
02:58:48,577 --> 02:58:50,146
INWARD AND OUTWARD CONFIRMATION 

3712
02:58:50,146 --> 02:58:51,847
FOR THIS -- FOR ANY PROTEIN, BUT

3713
02:58:51,847 --> 02:58:55,084
WE STARTED THIS BECAUSE WE HAD 

3714
02:58:55,084 --> 02:59:02,158
IDENTIFIED -- RERANDOMIZED THE 

3715
02:59:02,158 --> 02:59:04,827
RECEPTIVE BINDING DOMAINS OF 

3716
02:59:04,827 --> 02:59:06,662
LEUKEMIA VIRUSES, WE IDENTIFIED 

3717
02:59:06,662 --> 02:59:07,696
FUNCTIONAL ENVELOPES, AND WE 

3718
02:59:07,696 --> 02:59:09,899
IDENTIFIED THE RECEPTORS.

3719
02:59:09,899 --> 02:59:15,271
ONE OF THEM IS THIS SLC-14 

3720
02:59:15,271 --> 02:59:15,571
PROTEIN.

3721
02:59:15,571 --> 02:59:17,106
IT'S A RECEPTOR, AND WE STILL 

3722
02:59:17,106 --> 02:59:18,541
DON'T KNOW WHAT IT TRANSPORTS, 

3723
02:59:18,541 --> 02:59:20,009
BUT WE STARTED TO DO STRUCTURAL 

3724
02:59:20,009 --> 02:59:22,044
WORK, PREDICTIONS OF IT DURING 

3725
02:59:22,044 --> 02:59:23,112
COVID, AND NOW WE HAVE THIS 

3726
02:59:23,112 --> 02:59:24,413
METHOD THAT LOOKS PRETTY GOOD 

3727
02:59:24,413 --> 02:59:27,049
AND WE'RE ABLE TO PREDICT THE 

3728
02:59:27,049 --> 02:59:30,319
DIFFERENT CONFORMATIONS.

3729
02:59:30,319 --> 02:59:32,721
SO I WANT TO THANK ALAN FOR 

3730
02:59:32,721 --> 02:59:35,357
OPENING MY EYES TO AI BACK IN 

3731
02:59:35,357 --> 02:59:37,793
2015 WITH EX MACHINA, AND I JUST

3732
02:59:37,793 --> 02:59:38,561
APPRECIATE EVERYTHING HE'S DONE 

3733
02:59:38,561 --> 02:59:41,297
AS A RESEARCHER, MENTOR, FRIEND,

3734
02:59:41,297 --> 02:59:42,097
AND MOVIE CRITIC.

3735
02:59:42,097 --> 02:59:52,274
THANK YOU.

3736
03:00:04,420 --> 03:00:05,988
>> WHAT ARE YOU WATCHING
NOW?

3737
03:00:05,988 --> 03:00:08,491
>> THE GREAT BRITISH
BAKING 

3738
03:00:08,491 --> 03:00:12,428
SHOW EVERY FRIDAY.

3739
03:00:12,428 --> 03:00:13,629
>> WHO ARE YOU HOPING TO
WIN 

3740
03:00:13,629 --> 03:00:14,597
THE BAKEOFF?

3741
03:00:14,597 --> 03:00:17,099
>> I DO LIKE DYLAN.

3742
03:00:17,099 --> 03:00:27,343
>> HE'S CUTE.

3743
03:00:39,221 --> 03:00:41,390
>> THAT WAS ABOUT THE
NICEST 

3744
03:00:41,390 --> 03:00:46,729
TALK I'VE EVER HEARD.

3745
03:00:46,729 --> 03:00:47,062
THANK YOU.

3746
03:00:47,062 --> 03:00:50,466
SO OUR NEXT SPEAKER IS ONE OF A 

3747
03:00:50,466 --> 03:00:55,938
SERIES OF SPEAKERS WHO NEEDS NO 

3748
03:00:55,938 --> 03:00:56,272
INTRODUCTION.

3749
03:00:56,272 --> 03:00:59,341
IT'S STEVE GOFF, WHO HAS BEEN AT

3750
03:00:59,341 --> 03:01:08,751
COLUMBIA FOR PROBABLY A FEW 

3751
03:01:08,751 --> 03:01:09,018
DECADES.

3752
03:01:09,018 --> 03:01:10,619
HE AND I WERE BOTH INTERESTED IN

3753
03:01:10,619 --> 03:01:14,223
HOW RETROVIRUSES WORK BEFORE HIV

3754
03:01:14,223 --> 03:01:16,825
CAME ALONG, AND THEREFORE, WE 

3755
03:01:16,825 --> 03:01:20,362
BOTH BEGAN WORKING WITH MURINE 

3756
03:01:20,362 --> 03:01:22,598
LEUKEMIA VIRUSES.

3757
03:01:22,598 --> 03:01:26,902
I'LL JUST SAY IN 25 WORDS OR 

3758
03:01:26,902 --> 03:01:30,139
LESS THAT STEVE HAS REALLY FOUND

3759
03:01:30,139 --> 03:01:33,108
ONE AMAZING SURPRISE AFTER 

3760
03:01:33,108 --> 03:01:33,342
ANOTHER.

3761
03:01:33,342 --> 03:01:38,080
HE ALWAYS, LIKE MONICA STRANGELY

3762
03:01:38,080 --> 03:01:39,848
WAS SAYING ABOUT ME, HE ALWAYS 

3763
03:01:39,848 --> 03:01:41,250
COMES UP WITH SOMETHING QUITE 

3764
03:01:41,250 --> 03:01:42,885
NEW AND UNEXPECTED, CONNECTIONS 

3765
03:01:42,885 --> 03:01:45,354
BETWEEN THE VIRUSES THAT WE KNOW

3766
03:01:45,354 --> 03:01:47,456
A LOT ABOUT AND THE CELLS WHICH 

3767
03:01:47,456 --> 03:01:49,458
ARE STILL LARGELY BLACK BOXES, 

3768
03:01:49,458 --> 03:01:50,326
AT LEAST COMPARED TO THE 

3769
03:01:50,326 --> 03:01:57,967
VIRUSES.

3770
03:01:57,967 --> 03:01:59,602
I THINK I'LL JUST LEAVE IT AT 

3771
03:01:59,602 --> 03:01:59,868
THAT.

3772
03:01:59,868 --> 03:02:01,937
STEVE, THANK YOU FOR COMING.

3773
03:02:01,937 --> 03:02:03,105
I CAN'T IMAGINE WHAT YOU'RE 

3774
03:02:03,105 --> 03:02:04,473
GOING TO SAY, BUT I'M LOOKING 

3775
03:02:04,473 --> 03:02:14,717
FORWARD TO IT.

3776
03:02:23,993 --> 03:02:24,893
>> ALAN THANK YOU.

3777
03:02:24,893 --> 03:02:33,936
IT IS REALLY A PLEASURE.

3778
03:02:33,936 --> 03:02:35,437
LET'S GET STARTED.

3779
03:02:35,437 --> 03:02:37,806
I'M GOING TO DO TWO THINGS IN 

3780
03:02:37,806 --> 03:02:39,508
THE NEXT FEW MINUTES.

3781
03:02:39,508 --> 03:02:40,876
I'LL SPEND SOME TIME KIND OF 

3782
03:02:40,876 --> 03:02:42,978
DOING A LITTLE HISTORY, AND IF 

3783
03:02:42,978 --> 03:02:45,214
THERE'S AN IMPORTANT TAKE AWAY 

3784
03:02:45,214 --> 03:02:48,617
FROM THIS, IT'S HOW MUCH ALAN 

3785
03:02:48,617 --> 03:02:50,853
HAS BEEN A ROLE MODEL FOR ME, 

3786
03:02:50,853 --> 03:02:52,154
FOR EVERYTHING I'VE DONE HAS 

3787
03:02:52,154 --> 03:02:53,355
REALLY FOLLOWED IN ALAN'S FOOT 

3788
03:02:53,355 --> 03:03:03,198
STEPS.

3789
03:03:03,198 --> 03:03:04,833
THEN I'LL TRY TO GIVE YOU A 

3790
03:03:04,833 --> 03:03:05,734
BRIEF SCIENCE TALK ABOUT THINGS 

3791
03:03:05,734 --> 03:03:07,169
THAT ARE GOING ON IN THE LAB, 

3792
03:03:07,169 --> 03:03:09,872
WHICH I THINK ALAN WOULD LIKE TO

3793
03:03:09,872 --> 03:03:18,047
HAVE ALWAYS ALWAYS REAL SCIENCE 

3794
03:03:18,047 --> 03:03:18,313
BUILT IN.

3795
03:03:18,313 --> 03:03:28,624
THIS IS THE TRUTH.

3796
03:03:29,124 --> 03:03:32,461
THIS IS ALWAYS THE TRUTH BECAUSE

3797
03:03:32,461 --> 03:03:35,164
WE OFTEN HAVE THINGS THAT WE'RE 

3798
03:03:35,164 --> 03:03:36,165
INSPIRED BY BECAUSE WE WOULDN'T 

3799
03:03:36,165 --> 03:03:38,200
KNOW THIS IS A COOL TOPIC TO 

3800
03:03:38,200 --> 03:03:41,904
WORK ON IF IT WEREN'T FOR ALAN.

3801
03:03:41,904 --> 03:03:47,476
A FEW EXAMPLES ARE NC MUTANTS, 

3802
03:03:47,476 --> 03:03:52,748
THE GAG-POL READTHROUGH, 

3803
03:03:52,748 --> 03:03:54,216
MYRISTATE AND L DOMAINS.

3804
03:03:54,216 --> 03:03:55,517
ALL OF THESE YOU'VE HEARD ABOUT,

3805
03:03:55,517 --> 03:03:57,152
AND ALL OF THESE ALAN INSPIRED 

3806
03:03:57,152 --> 03:03:58,687
US TO WORK ON BECAUSE HE MADE 

3807
03:03:58,687 --> 03:04:00,723
SOME DISCOVERY ABOUT THESE.

3808
03:04:00,723 --> 03:04:02,191
I THINK LOOKING BACK AT THE 

3809
03:04:02,191 --> 03:04:03,859
HISTORY PART, THE FIRST TIME I 

3810
03:04:03,859 --> 03:04:05,661
MET ALAN MAYBE, YOU MAY CORRECT 

3811
03:04:05,661 --> 03:04:06,995
ME, WAS WHEN I WAS JUST STARTING

3812
03:04:06,995 --> 03:04:10,099
OUT AS A POST DOC IN DAVID 

3813
03:04:10,099 --> 03:04:11,200
BALTIMORE'S LAB.

3814
03:04:11,200 --> 03:04:13,202
THIS WOULD HAVE BEEN 1979, AND I

3815
03:04:13,202 --> 03:04:22,945
THINK HE WAS ON A SITE VISIT 

3816
03:04:22,945 --> 03:04:26,115
PANEL REVIEWING A GRANT, AND I 

3817
03:04:26,115 --> 03:04:27,983
THINK FOR SOME REASON HE HAD ME 

3818
03:04:27,983 --> 03:04:30,419
PRESENT A REVIEW STUDY TO THE 

3819
03:04:30,419 --> 03:04:30,619
PANEL.

3820
03:04:30,619 --> 03:04:32,154
SO I WAS TERRIFIED, OF COURSE.

3821
03:04:32,154 --> 03:04:35,357
THIS WAS ME JUST STARTING INTO 

3822
03:04:35,357 --> 03:04:37,259
RETROVIRUSES FROM SB-40.

3823
03:04:37,259 --> 03:04:39,862
SO OF COURSE I HAD READ ALL OF 

3824
03:04:39,862 --> 03:04:40,863
ALAN'S PAPERS AND WAS 

3825
03:04:40,863 --> 03:04:43,132
INTERESTED, ACTUALLY EVEN THEN, 

3826
03:04:43,132 --> 03:04:47,970
IN THINGS LIKE FV-1 WHICH 

3827
03:04:47,970 --> 03:04:49,872
INTRIGUED ME WITH MLV, AND THERE

3828
03:04:49,872 --> 03:04:52,241
WAS THIS GUY ALAN REIN, WHO I 

3829
03:04:52,241 --> 03:04:53,842
HAD TO PRESENT AND BE REVIEWED 

3830
03:04:53,842 --> 03:04:54,143
BY.

3831
03:04:54,143 --> 03:04:56,078
SO I DID, AND DAVID OF COURSE 

3832
03:04:56,078 --> 03:05:01,250
GOT THE GRANT, THANK YOU, BUT 

3833
03:05:01,250 --> 03:05:02,985
I'M SURE THROUGH NO VALUE OF 

3834
03:05:02,985 --> 03:05:03,952
ANYTHING I SAID BECAUSE I HAVE 

3835
03:05:03,952 --> 03:05:05,821
NO CLUE WHAT I SAID IN THAT 

3836
03:05:05,821 --> 03:05:09,091
PRESENTATION, BUT THERE WAS 

3837
03:05:09,091 --> 03:05:09,291
ALAN.

3838
03:05:09,291 --> 03:05:16,932
OVER THE YEARS SINCE THEN, I 

3839
03:05:16,932 --> 03:05:19,835
HAVE GOTTEN REAGENT AFTER 

3840
03:05:19,835 --> 03:05:21,270
REAGENT AFTER REAGENT FROM HIM, 

3841
03:05:21,270 --> 03:05:22,471
AND EVERY TIME HE SENDS ME 

3842
03:05:22,471 --> 03:05:24,072
EXACTLY WHAT I ASK FOR, AND IT'S

3843
03:05:24,072 --> 03:05:25,040
ALWAYS PERFECT AND IT'S JUST 

3844
03:05:25,040 --> 03:05:25,874
WHAT I NEEDED.

3845
03:05:25,874 --> 03:05:28,010
WHEN I STARTED OUT, THE FIRST 

3846
03:05:28,010 --> 03:05:31,213
THING I FELT I HAD TO DO WAS SET

3847
03:05:31,213 --> 03:05:32,447
UP A PLAQUE ASSAY BECAUSE THAT'S

3848
03:05:32,447 --> 03:05:34,783
WHAT I THOUGHT YOU NEEDED FOR 

3849
03:05:34,783 --> 03:05:35,050
VIROLOGY.

3850
03:05:35,050 --> 03:05:41,323
THAT IN THOSE DAYS WAS THE XE 

3851
03:05:41,323 --> 03:05:43,926
PLATH ASSAY, WHICH WAS A TRICKY 

3852
03:05:43,926 --> 03:05:45,928
ASSAY, AND ALAN SENT THE XE 

3853
03:05:45,928 --> 03:05:48,130
CELLS AROUND AND THE VERY TRICKY

3854
03:05:48,130 --> 03:05:49,798
PROTOCOL HOW TO DO IT, AND WE 

3855
03:05:49,798 --> 03:05:50,365
GOT GOING.

3856
03:05:50,365 --> 03:05:52,234
THAT WAS HOW WE STARTED OUT 

3857
03:05:52,234 --> 03:05:52,768
ASSAYING VIRUS.

3858
03:05:52,768 --> 03:05:54,403
IT WAS SO HARD THAT I WANTED 

3859
03:05:54,403 --> 03:05:56,839
BETTER ASSAYS, SO WE DID SET UP 

3860
03:05:56,839 --> 03:05:58,974
BETTER ASSAYS LATER THAT WERE 

3861
03:05:58,974 --> 03:05:59,308
EASIER.

3862
03:05:59,308 --> 03:06:01,710
BUT ALL OF THE CONCEPTS THAT 

3863
03:06:01,710 --> 03:06:11,687
FLOWED INTO MY WORK WERE THE 

3864
03:06:11,687 --> 03:06:14,389
VERY FIRST RECOMBINANT CLONES 

3865
03:06:14,389 --> 03:06:16,124
THAT ALLOWED YOU TO DO THINGS, 

3866
03:06:16,124 --> 03:06:17,960
AND ALAN AND I RAN WITH THAT AS 

3867
03:06:17,960 --> 03:06:19,494
FAR AS WE COULD, AND IT'S BEEN A

3868
03:06:19,494 --> 03:06:20,028
GOOD RIDE.

3869
03:06:20,028 --> 03:06:21,396
LET ME TALK SOME OF THE 

3870
03:06:21,396 --> 03:06:21,730
EXAMPLES.

3871
03:06:21,730 --> 03:06:23,065
YOU'VE SEEN SOME OF THIS BEFORE.

3872
03:06:23,065 --> 03:06:25,934
IN THE EARLY ABILITY TO MAKE 

3873
03:06:25,934 --> 03:06:27,469
MUTANTS AND CLONE DNA WAS WHAT 

3874
03:06:27,469 --> 03:06:28,937
ALLOWED FOR THE WORK TO SHOW 

3875
03:06:28,937 --> 03:06:31,206
THAT NC WAS REALLY IMPORTANT AND

3876
03:06:31,206 --> 03:06:32,808
WHAT IT DID AND HOW IT WAS 

3877
03:06:32,808 --> 03:06:35,010
INVOLVED IN PACKAGING, AND THIS 

3878
03:06:35,010 --> 03:06:37,479
WAS ONE OF THE BIG PAPERS FROM 

3879
03:06:37,479 --> 03:06:40,148
BOB AND ALAN, AND WE FOLLOWED 

3880
03:06:40,148 --> 03:06:41,783
CLOSELY UP, ALWAYS BEHIND OF 

3881
03:06:41,783 --> 03:06:44,052
COURSE, TO DO SIMILAR THINGS 

3882
03:06:44,052 --> 03:06:46,722
WITH CLAUDE TO LOOK AT NC 

3883
03:06:46,722 --> 03:06:48,824
MUTANTS.

3884
03:06:48,824 --> 03:06:50,025
WE KNEW IT WAS GOING TO BE A 

3885
03:06:50,025 --> 03:06:51,593
GOOD THING TO DO BECAUSE OF 

3886
03:06:51,593 --> 03:06:52,127
ALAN.

3887
03:06:52,127 --> 03:06:54,529
HERE'S ANOTHER ONE WHERE IN VERY

3888
03:06:54,529 --> 03:06:58,033
EARLY DAYS WITH STEVE OROSZLAN, 

3889
03:06:58,033 --> 03:07:00,502
BASED FIRST ON THE PROTEIN 

3890
03:07:00,502 --> 03:07:04,072
SEQUENCES REALLY, THAT THIS 

3891
03:07:04,072 --> 03:07:06,008
GAG-POL READTHROUGH THING HAD TO

3892
03:07:06,008 --> 03:07:08,944
BE IN FRAME AND INVOLVE 

3893
03:07:08,944 --> 03:07:12,080
READTHROUGH OF THE STOP CODON, 

3894
03:07:12,080 --> 03:07:12,447
AMAZING.

3895
03:07:12,447 --> 03:07:13,248
ONE OF THE THINGS THAT ALWAYS 

3896
03:07:13,248 --> 03:07:15,117
BLEW ME AWAY, THIS WAS NOT 

3897
03:07:15,117 --> 03:07:17,119
UNIVERSAL FOR ALL RETROVIRUSES, 

3898
03:07:17,119 --> 03:07:18,287
SOMETIMES THEY DID FRAME 

3899
03:07:18,287 --> 03:07:19,755
SHIFTING INSTEAD OF READTHROUGH,

3900
03:07:19,755 --> 03:07:21,390
THAT'S CRAZY, BUT THAT'S THE WAY

3901
03:07:21,390 --> 03:07:21,723
IT IS.

3902
03:07:21,723 --> 03:07:23,392
IT JUST SHOWS YOU THAT DIFFERENT

3903
03:07:23,392 --> 03:07:24,660
VIRUSES DO DIFFERENT THINGS, THE

3904
03:07:24,660 --> 03:07:27,029
SAME THING IN DIFFERENT WAYS.

3905
03:07:27,029 --> 03:07:28,297
WITH THAT, OF COURSE THEN YOU 

3906
03:07:28,297 --> 03:07:30,365
HAVE A CLONE, SO YOU CAN 

3907
03:07:30,365 --> 03:07:33,302
MANIPULATE THE NATURE OF THE 

3908
03:07:33,302 --> 03:07:40,475
READTHROUGH FEATURES, AND ALAN 

3909
03:07:40,475 --> 03:07:46,381
DID THAT WITH QD-11.

3910
03:07:46,381 --> 03:07:48,417
AND THERE WAS A READTHROUGH FOR 

3911
03:07:48,417 --> 03:07:50,452
THAT, AND THOSE WERE SEMINAL 

3912
03:07:50,452 --> 03:07:52,220
PAPERS, AND IN MY LAB THEY WERE 

3913
03:07:52,220 --> 03:07:54,022
ABLE TO FOLLOW UP AND DO MORE.

3914
03:07:54,022 --> 03:07:55,691
FROM FEELING YOUR WAY AROUND, 

3915
03:07:55,691 --> 03:07:57,592
LIKE FEELING THE ELEPHANT, YOU 

3916
03:07:57,592 --> 03:07:59,561
LEARN WHAT'S REQUIRED.

3917
03:07:59,561 --> 03:08:02,764
IN THE END, YEARS LATER, THAT IS

3918
03:08:02,764 --> 03:08:03,966
ANOTHER COMMON THING THAT YOU 

3919
03:08:03,966 --> 03:08:05,901
GET CONNECTED TO SERIOUS 

3920
03:08:05,901 --> 03:08:07,970
STRUCTURE PEOPLE WHO HELP YOU 

3921
03:08:07,970 --> 03:08:09,438
TRANS

3922
03:08:09,438 --> 03:08:11,506
TRANSLATE THE MUTATIONAL STUDY 

3923
03:08:11,506 --> 03:08:14,509
YOU DID TO TURN THE LIGHT ON AND

3924
03:08:14,509 --> 03:08:23,251
ACTUALLY SEE THE STRUCTURE, AND 

3925
03:08:23,251 --> 03:08:26,221
WE CONNECTED WITH PEOPLE LIKE 

3926
03:08:26,221 --> 03:08:29,024
VICKY AND MIKE, AND THAT BECOME 

3927
03:08:29,024 --> 03:08:34,596
THE PSEUDOKNOT, AND WE LEARNED 

3928
03:08:34,596 --> 03:08:36,398
WHICH OF THESE FEATURES ARE 

3929
03:08:36,398 --> 03:08:40,168
REALLY IMPORTANT.

3930
03:08:40,168 --> 03:08:42,938
IT HAS SOME AMAZING FEATURES.

3931
03:08:42,938 --> 03:08:44,673
IT TURNS OUT TO BE, THE 

3932
03:08:44,673 --> 03:08:45,974
STRUCTURE ITSELF, TURNS OUT TO 

3933
03:08:45,974 --> 03:08:48,944
BE INCREDIBLY PH DEPENDENT 

3934
03:08:48,944 --> 03:08:51,313
BECAUSE THERE'S A SINGLE 

3935
03:08:51,313 --> 03:08:52,447
PROTONAT

3936
03:08:52,447 --> 03:08:54,783
PROTONATED BASE THERE ON THE 

3937
03:08:54,783 --> 03:08:57,953
RIGHT THAT CHANGES THE STRUCTURE

3938
03:08:57,953 --> 03:08:59,821
COMPLETELY BECAUSE OF FUNCTION.

3939
03:08:59,821 --> 03:09:03,158
THAT'S A CONCEPT OF ENSEMBLES IN

3940
03:09:03,158 --> 03:09:05,293
OUR STRUCTURES THAT MY NEW 

3941
03:09:05,293 --> 03:09:09,698
COLLEAGUE AL HASHIMA MAKES FOR A

3942
03:09:09,698 --> 03:09:12,834
LIVING, AND THIS PSEUDOKNOT IS 

3943
03:09:12,834 --> 03:09:14,736
THE PERFECT EXAMPLE OF THAT.

3944
03:09:14,736 --> 03:09:15,737
THERE'S STILL TONS TO LEARN 

3945
03:09:15,737 --> 03:09:17,339
ABOUT HOW THIS WORKS, HOW IT 

3946
03:09:17,339 --> 03:09:18,874
INTERACTS WITH THE RIBOSOME, HOW

3947
03:09:18,874 --> 03:09:20,475
IT TELLS THE RIBOSOME TO DO ALL 

3948
03:09:20,475 --> 03:09:21,910
THE THINGS IT MUST BE TELLING IT

3949
03:09:21,910 --> 03:09:24,246
TO DO, BUT AGAIN THIS ALL 

3950
03:09:24,246 --> 03:09:31,953
STARTED WAY BACK WITH OROSZLAN 

3951
03:09:31,953 --> 03:09:34,022
AND ALAN'S PROTEIN SEQUENCE.

3952
03:09:34,022 --> 03:09:36,858
ANOTHER ONE IS THE WHOLE 

3953
03:09:36,858 --> 03:09:39,494
INSTALLATION STORY, THE VERY 

3954
03:09:39,494 --> 03:09:46,902
TERMINUS OF GAG.

3955
03:09:46,902 --> 03:09:53,275
THE GAG PROTEIN IS NOT 

3956
03:09:53,275 --> 03:09:54,643
MYRISTYLATABLE, SO THE GAG DOES 

3957
03:09:54,643 --> 03:09:57,012
NOT GET TO THE MEMBRANE AND DOES

3958
03:09:57,012 --> 03:09:59,181
NOT BUILD PARTICLES.

3959
03:09:59,181 --> 03:10:00,415
IT'S PROBABLY ONE OF THE 

3960
03:10:00,415 --> 03:10:01,349
STRONGEST PHENOTYPES FOR 

3961
03:10:01,349 --> 03:10:04,352
ANYTHING WITH A SINGLE AMINO 

3962
03:10:04,352 --> 03:10:05,020
ACID CHANGE.

3963
03:10:05,020 --> 03:10:06,321
THAT JUST CRYSTALLIZED US TO 

3964
03:10:06,321 --> 03:10:07,856
KEEP WORKING ON IT OVER THE 

3965
03:10:07,856 --> 03:10:08,056
YEARS.

3966
03:10:08,056 --> 03:10:09,724
ONE OF THE HOLY GRAILS WAS, 

3967
03:10:09,724 --> 03:10:12,828
WELL, IS THIS A PURELY 

3968
03:10:12,828 --> 03:10:20,502
SPONTANEOUS THING THAT A 

3969
03:10:20,502 --> 03:10:22,270
MYRISTYLATED PROTEIN IS PUT INTO

3970
03:10:22,270 --> 03:10:22,938
A CARRIER.

3971
03:10:22,938 --> 03:10:25,373
WE WENT LOOKING AND NEVER FIND A

3972
03:10:25,373 --> 03:10:28,143
CARRIER PROTEIN THAT DID IT, BUT

3973
03:10:28,143 --> 03:10:32,214
WE DID FIND BY PROTEOMICS, A 

3974
03:10:32,214 --> 03:10:33,715
BINDING PROTEIN.

3975
03:10:33,715 --> 03:10:36,418
IT HAS THE PERFECT POCKET, IT 

3976
03:10:36,418 --> 03:10:40,956
BINDS MYRISTASTE.

3977
03:10:40,956 --> 03:10:51,066
IT DOESN'T BIND NON-MYRISTATED  

3978
03:10:51,066 --> 03:10:52,167
PROTEINS.

3979
03:10:52,167 --> 03:10:54,136
IT'S PROBABLY GOT A BROAD ROLE, 

3980
03:10:54,136 --> 03:10:56,171
AND THAT CONNECTED US TO 

3981
03:10:56,171 --> 03:10:59,040
STRUCTURE PEOPLE SAYING, WELL, 

3982
03:10:59,040 --> 03:11:00,909
WE CAN CRYSTALLIZE THESE.

3983
03:11:00,909 --> 03:11:03,311
AND THERE'S THE MYRISTATE 

3984
03:11:03,311 --> 03:11:05,714
SITTING IN THIS POCKET.

3985
03:11:05,714 --> 03:11:11,653
ALL AMINO ACIDS MAKE CONTACT, 

3986
03:11:11,653 --> 03:11:14,689
AND THEY TOTALLY IMPACT THE 

3987
03:11:14,689 --> 03:11:16,224
CO-IT OF THESE PROTEINS THAT SIT

3988
03:11:16,224 --> 03:11:17,292
IN THIS BLOCK.

3989
03:11:17,292 --> 03:11:18,760
THESE ARE THE KINDS OF THREADS 

3990
03:11:18,760 --> 03:11:21,296
THAT ALL EMANATE FROM A STARTING

3991
03:11:21,296 --> 03:11:22,631
POINT.

3992
03:11:22,631 --> 03:11:26,835
I THINK THIS IS STILL RIPE FOR 

3993
03:11:26,835 --> 03:11:27,435
LOTS MORE WORK.

3994
03:11:27,435 --> 03:11:28,803
IT LOOKS LIKE, IF ANYTHING, THIS

3995
03:11:28,803 --> 03:11:30,438
IS PROBABLY NOT A CARRIER 

3996
03:11:30,438 --> 03:11:32,107
PROTEIN THAT DELIVERS IT TO THE 

3997
03:11:32,107 --> 03:11:32,374
MEMBRANE.

3998
03:11:32,374 --> 03:11:35,243
IT'S PROBABLY NOT A NEGATIVE 

3999
03:11:35,243 --> 03:11:39,147
REGULATOR OF MYRISTOYL 

4000
03:11:39,147 --> 03:11:41,449
FUNCTIONS, INCLUDING A KNOCKOUT 

4001
03:11:41,449 --> 03:11:43,818
WHERE THE CELLS MAKE THEM WAY 

4002
03:11:43,818 --> 03:11:45,420
BETTER AT ASSEMBLING THE 

4003
03:11:45,420 --> 03:11:46,755
PARTICLES, WHICH IS COOL.

4004
03:11:46,755 --> 03:11:49,057
THERE'S MORE AND MORE.

4005
03:11:49,057 --> 03:11:52,727
I HAVE TO SAY, IN SPITE OF ALL 

4006
03:11:52,727 --> 03:11:54,863
THESE OVERLAPS, ALAN AND I HAVE 

4007
03:11:54,863 --> 03:11:56,965
HARDLY EVER PUBLISHED TOGETHER.

4008
03:11:56,965 --> 03:12:00,869
THIS IS ONE OF THE FEW.

4009
03:12:00,869 --> 03:12:06,608
PEOPLE LIKE JOHN WILS AND OTHERS

4010
03:12:06,608 --> 03:12:09,177
DISCOVERED THE INTERCHANGE 

4011
03:12:09,177 --> 03:12:11,446
L-DOMAINS, THE FAMOUS SEQUENCES 

4012
03:12:11,446 --> 03:12:14,516
IN GAG THAT ARE NECESSARY TO 

4013
03:12:14,516 --> 03:12:14,950
EXCHANGE PARTICLES.

4014
03:12:14,950 --> 03:12:16,585
YOU COULD INTERCHANGE THEM, 

4015
03:12:16,585 --> 03:12:18,086
EXCHANGE THEM BETWEEN VIRUSES, 

4016
03:12:18,086 --> 03:12:19,654
AND WE FOUND THAT WAS TRUE WITH 

4017
03:12:19,654 --> 03:12:24,893
MLV, AND ALAN AND I GENERATED A 

4018
03:12:24,893 --> 03:12:26,661
WHOLE SERIES OF CONSTRUCTS THAT 

4019
03:12:26,661 --> 03:12:29,264
JUST GENERALIZED THAT PHENOMENON

4020
03:12:29,264 --> 03:12:31,866
TO MANY SITES FOR BINDING TO THE

4021
03:12:31,866 --> 03:12:36,938
MANY COMPONENTS OF THE S-CORE 

4022
03:12:36,938 --> 03:12:37,205
MACHINERY.

4023
03:12:37,205 --> 03:12:38,740
I'M VERY PROUD TO HAVE THAT ONE 

4024
03:12:38,740 --> 03:12:39,441
PAPER IN COMMON.

4025
03:12:39,441 --> 03:12:41,209
JUST TO WRAP UP THIS PART, I 

4026
03:12:41,209 --> 03:12:43,378
THINK IT'S IMPORTANT TO REFLECT 

4027
03:12:43,378 --> 03:12:46,548
ON WHY AND IN WHAT WAY ALAN HAS 

4028
03:12:46,548 --> 03:12:49,451
BEEN SO IMPORTANT TO US ALL, 

4029
03:12:49,451 --> 03:12:50,952
FIRST OF ALL, PROFESSIONALLY.

4030
03:12:50,952 --> 03:12:52,487
THE FIRST THING IS HE'S REVIEWED

4031
03:12:52,487 --> 03:12:55,257
MOST OF OUR PAPERS PROBABLY OF 

4032
03:12:55,257 --> 03:12:59,294
ALMOST EVERYBODY IN THE ROOM.

4033
03:12:59,294 --> 03:13:00,962
CRITICALLY AND VERY OFTEN 

4034
03:13:00,962 --> 03:13:02,464
POSITIVELY, THANKFULLY, AND HE'S

4035
03:13:02,464 --> 03:13:05,233
BEEN AN EDITOR OF ALL THE 

4036
03:13:05,233 --> 03:13:08,703
IMPORTANT VIROLOGY JOURNALS LIKE

4037
03:13:08,703 --> 03:13:09,571
JV AND VIROLOGY.

4038
03:13:09,571 --> 03:13:11,773
AS AN EDITOR, I USE HIM ALL THE 

4039
03:13:11,773 --> 03:13:13,408
TIME BECAUSE HE ALWAYS GIVES ME 

4040
03:13:13,408 --> 03:13:15,143
BACK FAIR AND ACCURATE REVIEWS 

4041
03:13:15,143 --> 03:13:16,611
OF ALL THE PAPERS THAT I GIVE 

4042
03:13:16,611 --> 03:13:17,812
HIM, WHICH IS GREAT.

4043
03:13:17,812 --> 03:13:21,049
ALAN RUNS THE BIG MEETINGS LIKE 

4044
03:13:21,049 --> 03:13:28,390
COLD SPRING HARBOR AND THE MANY 

4045
03:13:28,390 --> 03:13:33,328
NUCLEAR CAPSID SYMPOSIA THAT 

4046
03:13:33,328 --> 03:13:35,897
WILL BE RUN IN PRAGUE NEXT YEAR.

4047
03:13:35,897 --> 03:13:37,999
HE DOES GIVE AMAZING 

4048
03:13:37,999 --> 03:13:39,734
PRESENTATIONS, INCLUDING AT THE 

4049
03:13:39,734 --> 03:13:42,237
WEST COAST RETROVIRUS MEETING, 

4050
03:13:42,237 --> 03:13:45,040
AND AS ALREADY MENTIONED, AT THE

4051
03:13:45,040 --> 03:13:48,443
2016 TALK, WHICH IS A CAREER 

4052
03:13:48,443 --> 03:13:50,478
LIFETIME TALK, IT'S MENTIONED IN

4053
03:13:50,478 --> 03:13:52,514
LINKABLE, AND I WOULD ENCOURAGE 

4054
03:13:52,514 --> 03:13:54,082
EVERYBODY TO SEE HIS TALK 

4055
03:13:54,082 --> 03:13:55,583
BECAUSE IT'S REALLY A WALK DOWN 

4056
03:13:55,583 --> 03:13:56,851
THE LANE OF MLV AND THE HISTORY.

4057
03:13:56,851 --> 03:14:01,723
TERRIFIC.

4058
03:14:01,723 --> 03:14:03,191
NOW THAT'S ALL I WANTED TO SAY 

4059
03:14:03,191 --> 03:14:04,793
ABOUT MY MEMORIES OF THE TIME 

4060
03:14:04,793 --> 03:14:05,794
WITH ALAN.

4061
03:14:05,794 --> 03:14:08,096
I JUST WOULD END THAT PART AND 

4062
03:14:08,096 --> 03:14:09,664
JUST SAY THAT THE MOST CRITICAL 

4063
03:14:09,664 --> 03:14:11,666
THING TO ME IS HE'S ALWAYS THERE

4064
03:14:11,666 --> 03:14:13,101
WHENEVER I NEED SOMETHING, WHICH

4065
03:14:13,101 --> 03:14:13,835
IS OFTEN.

4066
03:14:13,835 --> 03:14:15,704
I CANNOT TELL YOU HOW MANY TIMES

4067
03:14:15,704 --> 03:14:18,340
I'VE ASKED HIM FOR PROBABLY THE 

4068
03:14:18,340 --> 03:14:25,814
SAME AMYTROPHIC GENOME, WHICH HE

4069
03:14:25,814 --> 03:14:26,715
SENDS TO ME.

4070
03:14:26,715 --> 03:14:27,916
HE CAN FIND THINGS IN HIS 

4071
03:14:27,916 --> 03:14:29,250
FREEZER AND SEND THEM TO ME, 

4072
03:14:29,250 --> 03:14:30,452
WHICH IS FABULOUS.

4073
03:14:30,452 --> 03:14:32,120
WE'VE SENT HIM A FEW THINGS, 

4074
03:14:32,120 --> 03:14:33,788
WHICH IS NOT AS MANY AS HE'S 

4075
03:14:33,788 --> 03:14:34,723
SENT TO US.

4076
03:14:34,723 --> 03:14:36,257
BEYOND THAT PROFESSIONAL THING, 

4077
03:14:36,257 --> 03:14:37,525
IT IS ALSO IMPORTANT TO SAY HE'S

4078
03:14:37,525 --> 03:14:39,661
BEEN IMPORTANT TO ME PERSONALLY 

4079
03:14:39,661 --> 03:14:41,062
AS A GOOD FRIEND.

4080
03:14:41,062 --> 03:14:43,365
I HAVE ALWAYS BEEN DELIGHTED TO 

4081
03:14:43,365 --> 03:14:46,101
BE ABLE TO MEET UP WITH HIM, 

4082
03:14:46,101 --> 03:14:47,669
USUALLY AT COLD SPRING HARBOR, 

4083
03:14:47,669 --> 03:14:50,138
AND DISCUSS EVERYTHING ABOUT OUR

4084
03:14:50,138 --> 03:14:52,407
CAREERS AND OUR THOUGHTS.

4085
03:14:52,407 --> 03:14:54,709
SO THANK YOU, ALAN.

4086
03:14:54,709 --> 03:14:56,511
LET ME TURN TO A QUICK LITTLE 

4087
03:14:56,511 --> 03:14:57,712
BIT OF SCIENCE BECAUSE I THINK 

4088
03:14:57,712 --> 03:14:59,114
IT'S A GOOD THING TO DO.

4089
03:14:59,114 --> 03:15:01,349
ONE OF THE PROJECTS GOING ON IN 

4090
03:15:01,349 --> 03:15:05,687
THE LAB IS ABOUT SUMOYLATION, SO

4091
03:15:05,687 --> 03:15:07,722
I'LL TELL YOU ABOUT THAT.

4092
03:15:07,722 --> 03:15:10,258
HOW DID I GET INTO THAT?

4093
03:15:10,258 --> 03:15:11,993
ACTUALLY, ONE OF THE WAYS I GOT 

4094
03:15:11,993 --> 03:15:13,895
INTO THIS HAD TO DO WITH 

4095
03:15:13,895 --> 03:15:16,264
JONATHAN A LITTLE BIT, WHICH WAS

4096
03:15:16,264 --> 03:15:17,866
IN CAPSID.

4097
03:15:17,866 --> 03:15:19,968
WE WERE LOOKING AT CAPSID 

4098
03:15:19,968 --> 03:15:20,902
MUTANTS, AND ONE OF THE THINGS 

4099
03:15:20,902 --> 03:15:22,404
WE WANTED TO KNOW IS WHAT ARE 

4100
03:15:22,404 --> 03:15:24,939
SOME OF THE PROTEINS THAT 

4101
03:15:24,939 --> 03:15:27,809
INTERACT WITH MLV CAPSID?

4102
03:15:27,809 --> 03:15:29,544
WE DID PROTEOMIC SCREENS AND GOT

4103
03:15:29,544 --> 03:15:30,745
A LOT OF HITS.

4104
03:15:30,745 --> 03:15:37,619
SOME OF THEM HAD TO DO WITH 

4105
03:15:37,619 --> 03:15:38,453
UBIQUITINYLATION AND 

4106
03:15:38,453 --> 03:15:39,320
SUMOYLATION.

4107
03:15:39,320 --> 03:15:40,054
WHICH IS ALL TRUE.

4108
03:15:40,054 --> 03:15:41,589
IT'S A BIG DEAL.

4109
03:15:41,589 --> 03:15:44,459
I THINK INCOMING CAPSIDES ARE 

4110
03:15:44,459 --> 03:15:46,094
RECOGNIZED BY THE 

4111
03:15:46,094 --> 03:15:48,229
UBIQUITINYLATION MACHINERY AND 

4112
03:15:48,229 --> 03:15:54,235
SOMETIMES SUMOYLATED, BUT I 

4113
03:15:54,235 --> 03:15:55,437
ALWAYS HAD THAT IN THE BACK OF 

4114
03:15:55,437 --> 03:15:57,105
MY HEAD TO FOLLOW UP ON.

4115
03:15:57,105 --> 03:15:58,273
ACTUALLY WITH JONATHAN IN THAT 

4116
03:15:58,273 --> 03:16:01,376
VEIN, WE DID A LOT OF TWO HYBRID

4117
03:16:01,376 --> 03:16:02,143
SCREENS.

4118
03:16:02,143 --> 03:16:05,480
WE DID CAPSID/CAPSID THAT 

4119
03:16:05,480 --> 03:16:06,247
INTERACTS NICELY.

4120
03:16:06,247 --> 03:16:08,817
WE DID A LOT OF CAPSID SCREENS 

4121
03:16:08,817 --> 03:16:11,653
THAT HAD TO DO WITH THESE GUYS 

4122
03:16:11,653 --> 03:16:15,423
WITH THE UBIQUITINYLATION, AND 

4123
03:16:15,423 --> 03:16:17,959
WE HAD FB-1, WHICH IS HORRIBLE 

4124
03:16:17,959 --> 03:16:20,161
BIOCHEMICALLY, BUT IT TURNS OUT 

4125
03:16:20,161 --> 03:16:22,430
TO BE WELL BEHAVED IN THE TWO 

4126
03:16:22,430 --> 03:16:23,364
HYBRID SCREEN SYSTEM.

4127
03:16:23,364 --> 03:16:25,867
IT'S GIVEN US LOTS OF HITS.

4128
03:16:25,867 --> 03:16:28,770
IT DIMERIZED BEAUTIFULLY IN THAT

4129
03:16:28,770 --> 03:16:31,039
SYSTEM, WHICH WE USED THROUGH 

4130
03:16:31,039 --> 03:16:33,241
MUTATIONS, AND IT INTERACTS WITH

4131
03:16:33,241 --> 03:16:41,483
THIS WHOLE UBUIQUITINE 

4132
03:16:41,483 --> 03:16:41,983
MACHINERY.

4133
03:16:41,983 --> 03:16:44,352
IT TURNS OUT SOMEDAY, IF WE'RE 

4134
03:16:44,352 --> 03:16:46,588
NOT CUT OFF FROM FUNDING, WE 

4135
03:16:46,588 --> 03:16:47,989
SHOULD LOOK AT THIS MORE.

4136
03:16:47,989 --> 03:16:51,826
SO WE DECIDED TO LOOK AT SUMO 

4137
03:16:51,826 --> 03:16:53,127
RECENTLY.

4138
03:16:53,127 --> 03:16:55,296
A NEW POST DOC LOOKED IT UP AND 

4139
03:16:55,296 --> 03:16:58,700
SAID WE'RE GOING TO DO A NEW 

4140
03:16:58,700 --> 03:17:00,635
PROTEOMIC SUMOYLATION WITH HIV.

4141
03:17:00,635 --> 03:17:03,304
IF YOU DON'T KNOW ALL ABOUT 

4142
03:17:03,304 --> 03:17:09,143
SUMO, IT'S A UBIQUITIN LIKE 

4143
03:17:09,143 --> 03:17:09,477
MODIFICATION.

4144
03:17:09,477 --> 03:17:14,015
IT CAN BE MONOSIMULATED OR 

4145
03:17:14,015 --> 03:17:14,282
MULTIPLE.

4146
03:17:14,282 --> 03:17:16,684
THERE ARE FIVE SUMOs IN MAMMALS 

4147
03:17:16,684 --> 03:17:17,252
AND PEOPLE.

4148
03:17:17,252 --> 03:17:20,221
THERE'S SUMO-1, WHICH IS 

4149
03:17:20,221 --> 03:17:21,322
DISTINCT.

4150
03:17:21,322 --> 03:17:23,625
SUMO-2 AND 3, WHICH ARE ALMOST 

4151
03:17:23,625 --> 03:17:24,392
IDENTICAL AND QUITE DIFFERENT 

4152
03:17:24,392 --> 03:17:28,663
FROM SUMO-1, AND DIFFERENT GUYS.

4153
03:17:28,663 --> 03:17:31,399
THE WAY THEY'RE PUT ON PROTEINS 

4154
03:17:31,399 --> 03:17:35,837
IS SIMILAR TO THE WAY UBIQUITIN 

4155
03:17:35,837 --> 03:17:37,772
IS PUT ON PROTEINS.

4156
03:17:37,772 --> 03:17:41,042
THERE'S LIGASES, E1, 2, AND 3, 

4157
03:17:41,042 --> 03:17:43,645
WHICH IS ALL IN PARALLEL TO WHAT

4158
03:17:43,645 --> 03:17:45,146
WE KNOW ABOUT UBIQUITIN.

4159
03:17:45,146 --> 03:17:47,615
THERE ARE MANY, MANY PROTEINS 

4160
03:17:47,615 --> 03:17:51,519
THAT ARE SUMOYLATED.

4161
03:17:51,519 --> 03:17:54,322
WHEN ERR, THEY'RE NOT TAGGED AT 

4162
03:17:54,322 --> 03:17:57,091
ALL THE WAY UBIQUITIN TAGS ARE.

4163
03:17:57,091 --> 03:18:01,162
THE MOST COMMON FEATURE THEY 

4164
03:18:01,162 --> 03:18:03,464
INTERMEDIATE IS THE DELIVERY 

4165
03:18:03,464 --> 03:18:05,233
INTO THE NUCLEUS, AND THEY HAVE 

4166
03:18:05,233 --> 03:18:08,102
THESE SUMO-INTERACTION MOTIFS, 

4167
03:18:08,102 --> 03:18:10,505
THESE SIMS, AND THEY DO A LOT OF

4168
03:18:10,505 --> 03:18:11,539
DIFFERENT THINGS IN THE CELL.

4169
03:18:11,539 --> 03:18:13,341
SO IT'S A BIG MODIFICATION.

4170
03:18:13,341 --> 03:18:14,909
THERE'S CERTAINLY A LOT OF 

4171
03:18:14,909 --> 03:18:16,945
OPPORTUNITIES IN THE HIV 

4172
03:18:16,945 --> 03:18:17,845
LIFECYCLE WHERE YOU COULD 

4173
03:18:17,845 --> 03:18:19,714
IMAGINE SUMO BEING IMPORTANT.

4174
03:18:19,714 --> 03:18:22,650
IT'S BEEN LOOKED AT IN VERY FEW 

4175
03:18:22,650 --> 03:18:23,284
CASES.

4176
03:18:23,284 --> 03:18:27,922
SO P6 IS SUMOYLATED, BUT IT 

4177
03:18:27,922 --> 03:18:29,791
DOESN'T SEEM TO DO ANYTHING.

4178
03:18:29,791 --> 03:18:35,630
SUMO IS A BIG REGULATOR OF NFAT,

4179
03:18:35,630 --> 03:18:38,433
AND THERE'S A LOT OF ROOM FOR 

4180
03:18:38,433 --> 03:18:40,702
SUMO TO BE DOING OTHER STUFF.

4181
03:18:40,702 --> 03:18:41,970
SO IN THE LAB THEY DECIDED TO 

4182
03:18:41,970 --> 03:18:44,439
LOOK AT IT IN A PROTEOMIC WAY.

4183
03:18:44,439 --> 03:18:46,708
THE PROBLEM WITH SUMO IN TERMS 

4184
03:18:46,708 --> 03:18:48,376
OF A PROTEOMIC SCREEN IS THAT 

4185
03:18:48,376 --> 03:18:52,981
THE USUAL WAY YOU LOOK AT 

4186
03:18:52,981 --> 03:18:55,183
UBIQUITINYLATION IS YOU DO 

4187
03:18:55,183 --> 03:18:57,418
REGULAR TRADITIONAL MASS SPEC, 

4188
03:18:57,418 --> 03:19:00,388
AND THE BEAUTY OF UBIQUITIN IS 

4189
03:19:00,388 --> 03:19:02,757
IT'S CLOSE TO THE C TERMINUS.

4190
03:19:02,757 --> 03:19:08,730
SO WHEN YOU MOVE YOUR LYSATE FOR

4191
03:19:08,730 --> 03:19:11,499
THE MASS SPEC, YOU LEAVE A 

4192
03:19:11,499 --> 03:19:17,572
LITTLE GLUSIN AT THE SITE OF THE

4193
03:19:17,572 --> 03:19:19,807
ADDITION, AND YOU CAN SEE IT AS 

4194
03:19:19,807 --> 03:19:22,210
A LITTLE BLIP IN THE PEPTIDE OF 

4195
03:19:22,210 --> 03:19:22,910
THE PRODUCT.

4196
03:19:22,910 --> 03:19:24,579
THAT'S GREAT.

4197
03:19:24,579 --> 03:19:27,949
UNFORTUNATELY, SUMO 1 AND SUMO 2

4198
03:19:27,949 --> 03:19:30,551
HAVE THE FIRST CLEAVABLE SITE 

4199
03:19:30,551 --> 03:19:32,520
NOWHERE NEAR THE C TERMINUS, AND

4200
03:19:32,520 --> 03:19:33,788
THE STUB THAT IS LEFT IS TOO BIG

4201
03:19:33,788 --> 03:19:35,289
TO ALLOW YOU TO IDENTIFY THE 

4202
03:19:35,289 --> 03:19:36,658
TARGET PROTEIN THAT WAY.

4203
03:19:36,658 --> 03:19:38,092
SO THE ANSWER, WHICH WE DIDN'T 

4204
03:19:38,092 --> 03:19:39,894
HAVE TO INVENT, IS THAT YOU 

4205
03:19:39,894 --> 03:19:42,830
CHANGE, YOU MAKE A SUMO 

4206
03:19:42,830 --> 03:19:45,199
MODIFIED, SUMO 1 OR SUMO 2 WITH 

4207
03:19:45,199 --> 03:19:50,872
A TRIPSIN CLEAVABLE AMINO ACID 

4208
03:19:50,872 --> 03:19:53,875
NEAR THE C TERMINUS, AND NOW THE

4209
03:19:53,875 --> 03:19:55,576
STUB IS SHORT ENOUGH TO SEE THIS

4210
03:19:55,576 --> 03:20:00,682
ON THE MASS SPEC PRODUCTS.

4211
03:20:00,682 --> 03:20:03,151
DMITRA MADE CELL LINES WITH 

4212
03:20:03,151 --> 03:20:04,152
THESE VARIANTS.

4213
03:20:04,152 --> 03:20:07,422
THEY'RE EXPRESSED AT THE 

4214
03:20:07,422 --> 03:20:08,990
ENDOGENOUS LEVELS, WHICH IS A 

4215
03:20:08,990 --> 03:20:09,657
BIG DEAL.

4216
03:20:09,657 --> 03:20:12,226
YOU'RE NOT OVEREXPRESSING 

4217
03:20:12,226 --> 03:20:14,262
ANYTHING.

4218
03:20:14,262 --> 03:20:15,863
THE MODIFIED SUMO BEHAVES WELL, 

4219
03:20:15,863 --> 03:20:17,231
GOES EVERYWHERE IT SHOULD GO, 

4220
03:20:17,231 --> 03:20:18,833
AND NOW YOU TAKE THOSE CELLS AND

4221
03:20:18,833 --> 03:20:20,368
ASK WHAT HAPPENS WHEN YOU INFECT

4222
03:20:20,368 --> 03:20:20,935
WITH HIV.

4223
03:20:20,935 --> 03:20:23,604
SO SHE DID THAT, SHE TOOK -- AND

4224
03:20:23,604 --> 03:20:26,441
SHE MADE VERSIONS, I SHOULD SAY,

4225
03:20:26,441 --> 03:20:29,477
THAT ARE FLAG TAGGED OR HIS 

4226
03:20:29,477 --> 03:20:30,645
TAGGED WHICH IS HELPFUL.

4227
03:20:30,645 --> 03:20:32,313
YOU HAVE THE CELL LINES READY TO

4228
03:20:32,313 --> 03:20:36,951
GO, INFECT THEM WITH HIV, MAKE 

4229
03:20:36,951 --> 03:20:37,185
LYSATES.

4230
03:20:37,185 --> 03:20:38,553
YOU CAN PULL THE TAGS OUT, WHICH

4231
03:20:38,553 --> 03:20:40,254
HELPS, YOU DON'T HAVE TO, AND 

4232
03:20:40,254 --> 03:20:41,789
SEND THE WHOLE DAMN THING OFF TO

4233
03:20:41,789 --> 03:20:43,257
THE MASS SPEC PEOPLE AND THEY 

4234
03:20:43,257 --> 03:20:45,460
COME BACK WITH A LIST OF 

4235
03:20:45,460 --> 03:20:47,028
EVERYTHING THAT'S SUMOYLATED.

4236
03:20:47,028 --> 03:20:50,098
AND WE ASK THOSE, OKAY, WHAT'S 

4237
03:20:50,098 --> 03:20:53,334
GONE UP WITH SUMOYLATION AND 

4238
03:20:53,334 --> 03:20:54,168
MAYBE WHAT'S GONE DOWN.

4239
03:20:54,168 --> 03:20:55,703
THERE YOU GO.

4240
03:20:55,703 --> 03:20:57,338
YOU GET A HUGE HIT LIST, A BUNCH

4241
03:20:57,338 --> 03:20:58,873
OF THINGS ARE UP, AND SOME 

4242
03:20:58,873 --> 03:20:59,674
THINGS ARE WAY UP.

4243
03:20:59,674 --> 03:21:02,310
IT'S REALLY SOME OF THEM ARE 

4244
03:21:02,310 --> 03:21:05,079
VERY STRIKINGLY INCREASED IN THE

4245
03:21:05,079 --> 03:21:06,314
SUMOYLATION UPON INFECTION.

4246
03:21:06,314 --> 03:21:07,982
A SMALLER NUMBER ARE DOWN, AND I

4247
03:21:07,982 --> 03:21:09,317
THINK THERE'S A TON OF STUFF 

4248
03:21:09,317 --> 03:21:11,919
HERE FOR US TO WORK ON FOR A 

4249
03:21:11,919 --> 03:21:13,121
LONG TIME, BUT THEY'RE 

4250
03:21:13,121 --> 03:21:13,421
INTERESTING.

4251
03:21:13,421 --> 03:21:16,457
THERE'S A LOT OF OVERLAP BETWEEN

4252
03:21:16,457 --> 03:21:18,826
SUMO 1 AND SUMO 2, BY THE WAY.

4253
03:21:18,826 --> 03:21:21,562
A LOT OF PROTEINS ARE SUMOYLATED

4254
03:21:21,562 --> 03:21:22,430
BY EITHER ONE.

4255
03:21:22,430 --> 03:21:24,265
THERE ARE SOME THAT ARE 

4256
03:21:24,265 --> 03:21:25,166
SIGNIFICANTLY ONLY SUMOYLATED BY

4257
03:21:25,166 --> 03:21:27,635
ONLY SUMO 1 OR ONLY SUMO 2.

4258
03:21:27,635 --> 03:21:29,070
LOTS OF THESE ARE INTERESTING.

4259
03:21:29,070 --> 03:21:30,104
NOW WHAT DO YOU DO?

4260
03:21:30,104 --> 03:21:31,038
YOU'VE GOT THIS LIST.

4261
03:21:31,038 --> 03:21:32,640
WELL, YOU LOOK AT THE LIST AND 

4262
03:21:32,640 --> 03:21:34,876
SAY, WHAT DO WE GOT HERE?

4263
03:21:34,876 --> 03:21:37,512
WHAT HAS COME, AND WHAT MIGHT BE

4264
03:21:37,512 --> 03:21:38,679
INTERESTING TO LOOK AT?

4265
03:21:38,679 --> 03:21:41,149
YOU ARE ENCOURAGED BY THE FACT 

4266
03:21:41,149 --> 03:21:47,355
THAT YOU SEND THESE SAMPLES TO 

4267
03:21:47,355 --> 03:21:49,857
MULTIPLE MASS SPEC FOR YOU, AND 

4268
03:21:49,857 --> 03:21:51,259
THEY COME UP WITH THE SAME THING

4269
03:21:51,259 --> 03:21:52,426
MOST OF THE TIME OR SOME OF THE 

4270
03:21:52,426 --> 03:21:53,661
TIME, AND THAT'S GOOD.

4271
03:21:53,661 --> 03:21:55,029
WE DID THAT.

4272
03:21:55,029 --> 03:21:57,298
SO WHAT DO WE GOT?

4273
03:21:57,298 --> 03:21:57,899
THERE WERE SOME INTERESTING 

4274
03:21:57,899 --> 03:22:00,368
THEMES THAT CAME UP IN THIS 

4275
03:22:00,368 --> 03:22:04,972
SCR

4276
03:22:04,972 --> 03:22:07,108
SCREEN, AND THE MOST STRIKING IS

4277
03:22:07,108 --> 03:22:17,018
THERE'S A WHOLE LOT OF HNRNPs.

4278
03:22:17,018 --> 03:22:19,654
HNRNP 1 AND 3, I'M GOING TO TALK

4279
03:22:19,654 --> 03:22:19,854
ABOUT.

4280
03:22:19,854 --> 03:22:21,656
THEY COME UP WITH SUMO 1 AND 

4281
03:22:21,656 --> 03:22:22,156
SUMO 2.

4282
03:22:22,156 --> 03:22:23,324
THESE ARE A LOT.

4283
03:22:23,324 --> 03:22:25,526
THESE ARE TWO LOGS AND MORE UP 

4284
03:22:25,526 --> 03:22:30,598
IN INFECTED CELLS, SO HUH, DON'T

4285
03:22:30,598 --> 03:22:31,265
KNOW.

4286
03:22:31,265 --> 03:22:32,733
THERE ARE OTHER INTERESTING 

4287
03:22:32,733 --> 03:22:33,434
THINGS.

4288
03:22:33,434 --> 03:22:39,273
THERE'S A LOT OF NUPs, NUCLEAR 

4289
03:22:39,273 --> 03:22:41,809
CORE PROTEINS, WHICH MAY BE 

4290
03:22:41,809 --> 03:22:42,176
INTERESTING.

4291
03:22:42,176 --> 03:22:43,644
BUT WE'RE LOOKING AT THOSE GUYS.

4292
03:22:43,644 --> 03:22:45,813
EVEN WHEN YOU LOOK KIND OF IN 

4293
03:22:45,813 --> 03:22:47,114
THE GO CATEGORIZATION OF WHAT 

4294
03:22:47,114 --> 03:22:50,251
THINGS ARE INCREASED, YOU REALLY

4295
03:22:50,251 --> 03:22:53,955
GET A LOT OF RNA HANDLING GUYS, 

4296
03:22:53,955 --> 03:22:58,092
RNA SPLICING GUYS LIKE THE RNPs,

4297
03:22:58,092 --> 03:23:00,027
A LOT OF RNA PROCESSING GUYS, A 

4298
03:23:00,027 --> 03:23:01,529
LOT OF GENES FLAGGED HAVING TO 

4299
03:23:01,529 --> 03:23:03,531
DO WITH THE METABOLISM OF RNA.

4300
03:23:03,531 --> 03:23:05,466
I SERIOUSLY BELIEVE IN INFECTION

4301
03:23:05,466 --> 03:23:07,468
THE VIRUS IS SCREWING AROUND IN 

4302
03:23:07,468 --> 03:23:08,936
VARIOUS WAYS WITH THIS RNA 

4303
03:23:08,936 --> 03:23:09,203
MACHINERY.

4304
03:23:09,203 --> 03:23:11,539
SO THESE ARE THE DOTS FOR THOSE 

4305
03:23:11,539 --> 03:23:18,646
TWO GENES, THOSE PARTICULAR GUYS

4306
03:23:18,646 --> 03:23:25,686
THIS IS THE AVERAGE OF 

4307
03:23:25,686 --> 03:23:27,521
TRIPLICATES BY THE MASS SPEC 

4308
03:23:27,521 --> 03:23:27,722
GUYS.

4309
03:23:27,722 --> 03:23:29,323
NOW WE CAN GO BACK AND LOOK AT 

4310
03:23:29,323 --> 03:23:31,292
ACTUAL PROTEIN WORKUPS, WHICH IS

4311
03:23:31,292 --> 03:23:31,525
TRUE.

4312
03:23:31,525 --> 03:23:34,762
IF YOU LOOK AT UNINFECTED AND 

4313
03:23:34,762 --> 03:23:36,597
INFECTED FOR SUMO 1 FOR CONTROL,

4314
03:23:36,597 --> 03:23:37,698
YOU HAVE YOUR CONTROLS, AND WHAT

4315
03:23:37,698 --> 03:23:41,669
YOU SEE IS THE PROTEIN IS HUGELY

4316
03:23:41,669 --> 03:23:43,170
SUMOYLATED AFTER INFECTION.

4317
03:23:43,170 --> 03:23:46,274
THAT UPPER BAND WITH THE ARROW 

4318
03:23:46,274 --> 03:23:47,942
IS MONOSUMOYLATED.

4319
03:23:47,942 --> 03:23:49,076
THE UNSUMOYLATED DOESN'T CHANGE.

4320
03:23:49,076 --> 03:23:51,045
THE PROTEIN LEVEL IS NOT 

4321
03:23:51,045 --> 03:23:51,712
AFFECTED.

4322
03:23:51,712 --> 03:23:54,215
WE HAVE THESE ADDED BANDS, 

4323
03:23:54,215 --> 03:24:00,321
PROBABLY THOSE ARE DI AND 

4324
03:24:00,321 --> 03:24:02,189
TRI-SUMOYLATED VERSIONS, PRETTY 

4325
03:24:02,189 --> 03:24:02,590
DRAMATIC.

4326
03:24:02,590 --> 03:24:04,926
JUST TO REMIND YOU WHAT'S 

4327
03:24:04,926 --> 03:24:06,060
POSSIBLE, WHICH DON'T KNOW WHICH

4328
03:24:06,060 --> 03:24:07,895
IS HAPPENING, AND WE DON'T KNOW 

4329
03:24:07,895 --> 03:24:09,430
ALL THE PLACES WHERE THOSE 

4330
03:24:09,430 --> 03:24:09,997
MODIFICATIONS ARE PROBABLY 

4331
03:24:09,997 --> 03:24:19,006
HAPPENING.

4332
03:24:19,006 --> 03:24:21,142
THEN IF YOU EXPOSE IT MORE, YOU 

4333
03:24:21,142 --> 03:24:23,878
CAN MAKE IT LOOK EVEN MORE 

4334
03:24:23,878 --> 03:24:24,178
BETTER.

4335
03:24:24,178 --> 03:24:25,546
YOU CAN MAKE THE FACTORS LOOK 

4336
03:24:25,546 --> 03:24:26,714
THE WAY YOU WANT THEM.

4337
03:24:26,714 --> 03:24:29,283
FOR SUMO 1, IT'S TRUE.

4338
03:24:29,283 --> 03:24:33,888
FOR HNRNPA3.

4339
03:24:33,888 --> 03:24:36,891
THERE'S A BIG BASAL LEVEL 

4340
03:24:36,891 --> 03:24:38,025
SUMOYLATION, BUT THEN THEN 

4341
03:24:38,025 --> 03:24:39,994
THERE'S A BIG INCREASE AFTER 

4342
03:24:39,994 --> 03:24:40,261
INFECTION.

4343
03:24:40,261 --> 03:24:44,498
YOU CAN SEE THAT OVER THERE.

4344
03:24:44,498 --> 03:24:46,667
AND WE KNOW WE CAN SHOW THAT AT 

4345
03:24:46,667 --> 03:24:50,471
LEAST THESE BANDS ARE SUMO 

4346
03:24:50,471 --> 03:24:52,206
ADDITION BECAUSE THERE'S A 

4347
03:24:52,206 --> 03:24:55,476
DYNAMITE SUMO INHIBITOR.

4348
03:24:55,476 --> 03:24:57,244
TACK THE DRUG AND THROW IT ON, 

4349
03:24:57,244 --> 03:24:58,679
AND IT TOTALLY BLOCKS THAT 

4350
03:24:58,679 --> 03:25:00,748
CHANGE AFTER INFECTION.

4351
03:25:00,748 --> 03:25:03,517
IT'S CLEAR THAT THIS IS 

4352
03:25:03,517 --> 03:25:04,885
SUMOYLATION.

4353
03:25:04,885 --> 03:25:05,853
SO THAT WAS INTERESTING.

4354
03:25:05,853 --> 03:25:07,922
YOU KNOW, WE DON'T KNOW REALLY A

4355
03:25:07,922 --> 03:25:08,789
WHOLE LOT MORE YET.

4356
03:25:08,789 --> 03:25:10,925
WHAT WE KNOW IS THE TIMING.

4357
03:25:10,925 --> 03:25:12,026
IT'S JUST KIND OF INTERESTING.

4358
03:25:12,026 --> 03:25:13,761
IT'S NOT EARLY IN INFECTION.

4359
03:25:13,761 --> 03:25:15,096
IT'S QUITE LATE.

4360
03:25:15,096 --> 03:25:17,164
SO HERE WE'VE LOOKED AT 

4361
03:25:17,164 --> 03:25:18,132
DIFFERENT TIMES AFTER 

4362
03:25:18,132 --> 03:25:22,069
INFECTION -- 6, 12, 24, 48 -- 

4363
03:25:22,069 --> 03:25:24,138
AND IT REALLY KICKS IN ONLY 

4364
03:25:24,138 --> 03:25:26,540
AROUND 24 HOURS AFTER INFECTION.

4365
03:25:26,540 --> 03:25:28,175
SO IT'S PROBABLY REQUIRING A 

4366
03:25:28,175 --> 03:25:28,676
VIRAL INTEGRATION AND 

4367
03:25:28,676 --> 03:25:33,180
EXPRESSION.

4368
03:25:33,180 --> 03:25:35,683
SO SOME LATE PROTEIN, SOME 

4369
03:25:35,683 --> 03:25:36,484
ASPECT RELATIVELY LATE IN 

4370
03:25:36,484 --> 03:25:39,487
INFECTION IS INDUCING THIS BIG 

4371
03:25:39,487 --> 03:25:41,956
MODIFICATION OF THE HNRNPs, AND 

4372
03:25:41,956 --> 03:25:44,492
THAT'S TRUE OF BOTH OF THOSE TWO

4373
03:25:44,492 --> 03:25:52,633
PARTICULAR PROTEINS.

4374
03:25:52,633 --> 03:25:54,969
IT'S NOT ONLY HELA CELLS WHERE 

4375
03:25:54,969 --> 03:25:57,505
WE'VE DONE THE MOST, BUT IT'S 

4376
03:25:57,505 --> 03:26:00,508
TRUE IN CELLS, AND IT SEEMS TO 

4377
03:26:00,508 --> 03:26:02,276
BE QUITE GENERAL.

4378
03:26:02,276 --> 03:26:05,046
WE'LL BE ASKED IN PRIMARY 

4379
03:26:05,046 --> 03:26:06,514
T-CELLS ANY DAY NOW, BUT WE 

4380
03:26:06,514 --> 03:26:08,349
HAVEN'T DONE THAT, BUT I BET IT 

4381
03:26:08,349 --> 03:26:09,517
WILL BE TRUE BECAUSE IT SEEMS TO

4382
03:26:09,517 --> 03:26:10,484
BE TRUE FOR BOTH.

4383
03:26:10,484 --> 03:26:13,287
WHAT ARE THESE PROTEINS DOING?

4384
03:26:13,287 --> 03:26:15,923
THEY'RE KNOWN TO BE HELPFUL IN 

4385
03:26:15,923 --> 03:26:17,324
SPLICING, AND THAT'S WHERE WE'LL

4386
03:26:17,324 --> 03:26:17,525
GO.

4387
03:26:17,525 --> 03:26:18,926
WE'VE DONE A LITTLE WORK ON 

4388
03:26:18,926 --> 03:26:21,829
THAT, SO YOU ALL KNOW IT'S 

4389
03:26:21,829 --> 03:26:22,129
COMPLICATED.

4390
03:26:22,129 --> 03:26:24,832
THERE'S TWO DONORS, ALL THESE 

4391
03:26:24,832 --> 03:26:27,068
SPLICE ACCEPTORS, AND THE RESULT

4392
03:26:27,068 --> 03:26:29,336
IS A MESS OF SPLICING.

4393
03:26:29,336 --> 03:26:31,005
THERE'S ALWAYS OFTEN THIS FIRST 

4394
03:26:31,005 --> 03:26:33,340
DONOR TO A BUNCH OF ACCEPTORS, 

4395
03:26:33,340 --> 03:26:34,975
AND THEN THERE'S THIS SET OTHER 

4396
03:26:34,975 --> 03:26:37,344
DONOR THAT CAN BE USED OR NOT TO

4397
03:26:37,344 --> 03:26:39,213
ITS ACCEPTOR, AND THERE'S 

4398
03:26:39,213 --> 03:26:41,415
ALTERNATE ACCEPTORS OVER THERE 

4399
03:26:41,415 --> 03:26:43,751
THAT MAKE MORE ALTERNATE 

4400
03:26:43,751 --> 03:26:46,554
PROTE

4401
03:26:46,554 --> 03:26:46,821
PROTEINS.

4402
03:26:46,821 --> 03:26:49,156
SO WHEN YOU ADD UP ALL THESE 

4403
03:26:49,156 --> 03:26:51,258
COMBOS, THERE'S AT LEAST 50 OR 

4404
03:26:51,258 --> 03:26:53,461
MORE SPLICE VARIANT MRNAs THAT 

4405
03:26:53,461 --> 03:26:56,163
ARE FORMED ON THE VIRUS.

4406
03:26:56,163 --> 03:26:58,499
THE RATIO OF ALL OF THEM ARE 

4407
03:26:58,499 --> 03:27:00,534
PRETTY STEREOTYPICAL ACTUALLY, 

4408
03:27:00,534 --> 03:27:02,203
BUT I'M SURE UNDER COMPLICATED 

4409
03:27:02,203 --> 03:27:03,671
CONTROL, AND I DON'T THINK WE 

4410
03:27:03,671 --> 03:27:04,772
REALLY KNOW MUCH ABOUT IT.

4411
03:27:04,772 --> 03:27:06,407
A LITTLE IS BEING KNOWN ABOUT 

4412
03:27:06,407 --> 03:27:08,843
THE SITES OF THE ACCEPTORS AND 

4413
03:27:08,843 --> 03:27:10,478
THERE ARE NOW A FAIR NUMBER OF 

4414
03:27:10,478 --> 03:27:12,246
MUTANTS THAT FOOL AROUND WITH 

4415
03:27:12,246 --> 03:27:14,248
THE ACCEPTOR USE.

4416
03:27:14,248 --> 03:27:18,285
I THINK OF THE -- A LOT OF THAT 

4417
03:27:18,285 --> 03:27:21,088
WORK BEING DONE BY ANN EMERY 

4418
03:27:21,088 --> 03:27:27,561
DOWN THERE AND RON SWANSTROM'S 

4419
03:27:27,561 --> 03:27:27,761
LAB.

4420
03:27:27,761 --> 03:27:30,064
WE HOOKED UP WITH HER AND 

4421
03:27:30,064 --> 03:27:31,665
KNOCKED DOWN OUR TWO FAVORITE 

4422
03:27:31,665 --> 03:27:33,968
SUMOYLATED PROTEINS, AND THAT'S 

4423
03:27:33,968 --> 03:27:34,468
WHAT HAPPENS.

4424
03:27:34,468 --> 03:27:36,570
SHE DOES THE ANALYSIS BY DEEP 

4425
03:27:36,570 --> 03:27:37,705
SEQUENCING WITH RANDOM PRIMERS 

4426
03:27:37,705 --> 03:27:40,641
UP ON TOP, AND SHE DOES IT WITH 

4427
03:27:40,641 --> 03:27:42,276
SPECIFIC PRIMERS THAT READ OUT 

4428
03:27:42,276 --> 03:27:43,811
INDIVIDUAL SPLICE SITES.

4429
03:27:43,811 --> 03:27:45,613
I WON'T WALK THROUGH IT EXCEPT 

4430
03:27:45,613 --> 03:27:47,548
TO SAY THERE ARE PRETTY BIG 

4431
03:27:47,548 --> 03:27:49,950
EFFECTS, SO WHEN YOU KNOCK DOWN 

4432
03:27:49,950 --> 03:27:52,453
SOME OF THE HNRNPs, JUST LIKE 

4433
03:27:52,453 --> 03:27:55,089
OUR FAVORITE GUYS, WE SEE THESE 

4434
03:27:55,089 --> 03:27:58,959
OFTEN BIG INCREASES IN SPLICE 

4435
03:27:58,959 --> 03:28:01,295
USAGE OF SOME OF THE PARTICULAR 

4436
03:28:01,295 --> 03:28:02,463
SPLICE, PARTIALLY SPLICED SITES 

4437
03:28:02,463 --> 03:28:07,735
UP ON THAT TOP RIGHT.

4438
03:28:07,735 --> 03:28:10,304
AND BIG CHANGES IN PARTICULAR 

4439
03:28:10,304 --> 03:28:13,641
SPLICES, USES OF ACCEPTOR 2, 

4440
03:28:13,641 --> 03:28:15,242
ACCEPTOR 3 WHEN YOU KNOCK THOSE 

4441
03:28:15,242 --> 03:28:15,476
DOWN.

4442
03:28:15,476 --> 03:28:16,977
THE FUTURE IS CLEAR OF WHAT WE 

4443
03:28:16,977 --> 03:28:19,780
WILL BE WANTING TO DO, WHICH IS 

4444
03:28:19,780 --> 03:28:24,385
TRY TO UNDERSTAND IF SUMOYLATION

4445
03:28:24,385 --> 03:28:27,488
IS A DEAL IN THIS, AND IF THAT'S

4446
03:28:27,488 --> 03:28:29,156
PART OF WHAT THE VIRUS IS DOING 

4447
03:28:29,156 --> 03:28:30,624
TO CONTROL ITS OWN SPLICING.

4448
03:28:30,624 --> 03:28:33,961
WE'RE GOING TO ASK DOES THE 

4449
03:28:33,961 --> 03:28:35,362
HIV-INDUCED SUMOYLATION WHICH WE

4450
03:28:35,362 --> 03:28:37,798
KNOW IS HAPPENING REALLY 

4451
03:28:37,798 --> 03:28:39,633
REGULATING THE FUNCTIONALITY OF 

4452
03:28:39,633 --> 03:28:41,435
THESE GUYS OR IS IT JUST 

4453
03:28:41,435 --> 03:28:41,735
HAPPENING?

4454
03:28:41,735 --> 03:28:44,338
AND WE'LL FIND OUT BECAUSE WE'RE

4455
03:28:44,338 --> 03:28:52,913
GOING TO MAKE NON-SUMOYLATEDABLE

4456
03:28:52,913 --> 03:28:54,582
MUTANTS OF THE PROTEINS AND SEE 

4457
03:28:54,582 --> 03:28:55,749
WHAT'S ADDED.

4458
03:28:55,749 --> 03:28:58,052
BUT SPLICING AND VIRUS 

4459
03:28:58,052 --> 03:28:58,385
REPLICATION.

4460
03:28:58,385 --> 03:29:00,287
I'LL STOP THERE AND THANK ALAN 

4461
03:29:00,287 --> 03:29:00,521
AGAIN.

4462
03:29:00,521 --> 03:29:02,856
HERE'S THE CURRENT GANG.

4463
03:29:02,856 --> 03:29:08,896
I THINK THERE ARE SO MANY 

4464
03:29:08,896 --> 03:29:09,763
AVENUES OF RESEARCH WE HAVE YET 

4465
03:29:09,763 --> 03:29:11,298
TO RUN WITH, SO MANY OF WHICH 

4466
03:29:11,298 --> 03:29:15,302
ARE STARTED BY ALAN.

4467
03:29:15,302 --> 03:29:17,705
WE'LL KEEP RUNNING AS LONG AS WE

4468
03:29:17,705 --> 03:29:17,871
CAN.

4469
03:29:17,871 --> 03:29:18,105
THANKS.

4470
03:29:18,105 --> 03:29:28,382
I'LL STOP THERE.

4471
03:29:35,055 --> 03:29:37,057
>> I'M SURE YOU'RE IN THE 

4472
03:29:37,057 --> 03:29:39,593
PRESENCE OF FINDING OUT WHICH OF

4473
03:29:39,593 --> 03:29:41,161
THESE RETROVIRAL PROTEINS 

4474
03:29:41,161 --> 03:29:41,996
CONTRIBUTE TO THIS?

4475
03:29:41,996 --> 03:29:42,997
>> I DON'T.

4476
03:29:42,997 --> 03:29:46,267
THE CHEAP WAY, WE'RE GOING TO 

4477
03:29:46,267 --> 03:29:48,168
OVEREXPRESS EACH ONE FIRST 

4478
03:29:48,168 --> 03:29:49,737
ALONE, NO VIRUS OTHERWISE, AND 

4479
03:29:49,737 --> 03:29:51,505
WE'LL SEE IF THAT'S IT.

4480
03:29:51,505 --> 03:29:52,973
AND THEN OF COURSE WE HAVE VIRUS

4481
03:29:52,973 --> 03:29:55,509
RESTRAINTS THAT LACK, YOU NAME 

4482
03:29:55,509 --> 03:29:58,646
IT, BPU, BPR, WHATEVER, WE'LL 

4483
03:29:58,646 --> 03:29:59,179
FIND OUT.

4484
03:29:59,179 --> 03:30:02,216
>> IT'S A BLUNT TOOL, IF
YOU 

4485
03:30:02,216 --> 03:30:05,152
LOOKED AT WHAT THE SUMOYLATION 

4486
03:30:05,152 --> 03:30:09,590
INHIBITORS DO TO HIV-1?

4487
03:30:09,590 --> 03:30:11,792
>> YES, AND THE ANSWER IS 

4488
03:30:11,792 --> 03:30:12,092
COMPLICATED.

4489
03:30:12,092 --> 03:30:13,627
THOSE DRUGS ARE FABULOUSLY 

4490
03:30:13,627 --> 03:30:15,829
POTENT, AND THEY ARE TOTALLY 

4491
03:30:15,829 --> 03:30:20,401
BLOCKING THE VIRUS, BUT THEY'RE 

4492
03:30:20,401 --> 03:30:21,802
PROBABLY DOING MORE THAN ONE 

4493
03:30:21,802 --> 03:30:22,002
THING.

4494
03:30:22,002 --> 03:30:24,805
THEY DO, AS I SHOWED YOU, BLOCK 

4495
03:30:24,805 --> 03:30:26,340
THE SUMOYLATION OF THESE 

4496
03:30:26,340 --> 03:30:29,610
FACTORS, YES, LATE IN INFECTION,

4497
03:30:29,610 --> 03:30:31,178
BUT THEIR BIG BLOCK, IS IT 

4498
03:30:31,178 --> 03:30:31,679
SOMETHING ELSE?

4499
03:30:31,679 --> 03:30:36,283
IT'S SUPER EARLY, AND IN FACT, 

4500
03:30:36,283 --> 03:30:37,685
THEIR MOST POTENT FUNCTION 

4501
03:30:37,685 --> 03:30:39,019
REQUIRES THE PRETREATMENT OF THE

4502
03:30:39,019 --> 03:30:40,487
CELLS FOR LIKE 12 HOURS BEFORE 

4503
03:30:40,487 --> 03:30:42,656
YOU INFECT THEM.

4504
03:30:42,656 --> 03:30:47,461
AND IF YOU DO THAT BEFORE AN 

4505
03:30:47,461 --> 03:30:48,662
INFECTION IS THREE LOGS DOWN, 

4506
03:30:48,662 --> 03:30:50,531
WHOPPING, BUT I GUESS IT'S NOT 

4507
03:30:50,531 --> 03:30:51,031
THOSE PROTEINS.

4508
03:30:51,031 --> 03:30:53,100
>> AND THE CELLS ARE FINE?

4509
03:30:53,100 --> 03:30:56,203
>> THEY DON'T LOVE IT, BUT

4510
03:30:56,203 --> 03:30:58,906
THEY'RE OKAY FOR -- THEY'RE NOT 

4511
03:30:58,906 --> 03:30:59,239
DEAD.

4512
03:30:59,239 --> 03:31:01,308
THEY'RE OKAY FOR A FEW DAYS, AND

4513
03:31:01,308 --> 03:31:03,577
THEY DON'T KNOW THEY'RE DEAD 

4514
03:31:03,577 --> 03:31:04,411
YET.

4515
03:31:04,411 --> 03:31:05,679
(LAUGHTER) 

4516
03:31:05,679 --> 03:31:07,848
AND THE INFECTION, THE VIRUS IS 

4517
03:31:07,848 --> 03:31:11,018
STILL ABLE TO DO THINGS IN THOSE

4518
03:31:11,018 --> 03:31:16,990
CELLS, YEAH.

4519
03:31:16,990 --> 03:31:20,427
>> SO IN TERMS OF THE 

4520
03:31:20,427 --> 03:31:23,097
SUPPRESSING BY INTERACTING WITH 

4521
03:31:23,097 --> 03:31:28,369
THE SERUM, I DON'T KNOW WHETHER 

4522
03:31:28,369 --> 03:31:37,678
YOU DEMONSTRATE THIS FACT?

4523
03:31:37,678 --> 03:31:40,681
>> YES, THAT'S THE $64,000

4524
03:31:40,681 --> 03:31:40,948
QUESTION.

4525
03:31:40,948 --> 03:31:42,249
EVERYBODY WOULD AGREE THEY'RE 

4526
03:31:42,249 --> 03:31:43,617
IMPORTANT IN THAT PROCESS 

4527
03:31:43,617 --> 03:31:45,719
SOMEHOW, AS REGULATORS OF 

4528
03:31:45,719 --> 03:31:46,620
SPLICING, AND WHETHER THE 

4529
03:31:46,620 --> 03:31:47,788
SUMOYLATION OF THEM IS GOOD OR 

4530
03:31:47,788 --> 03:31:49,857
BAD FOR THAT, WE DON'T KNOW.

4531
03:31:49,857 --> 03:31:52,126
THAT'S WHAT WE NEED TO FIND OUT.

4532
03:31:52,126 --> 03:32:00,701
I HOPE I'LL TELL YOU SOON.

4533
03:32:00,701 --> 03:32:06,607
>> ONE OF THE THINGS THAT
SEEM 

4534
03:32:06,607 --> 03:32:09,676
TO BE UPREGULATED FOR 

4535
03:32:09,676 --> 03:32:11,912
SUMOYLATION IS WHAT'S SITTING ON

4536
03:32:11,912 --> 03:32:13,680
THE EXIT TUNNEL.

4537
03:32:13,680 --> 03:32:15,516
HAVE YOU LOOKED TO SEE ABOUT 

4538
03:32:15,516 --> 03:32:15,849
TRANSLATIONAL?

4539
03:32:15,849 --> 03:32:16,550
>> WE HAVEN'T.

4540
03:32:16,550 --> 03:32:17,484
AND ONE OF THE GREAT THINGS 

4541
03:32:17,484 --> 03:32:19,019
ABOUT PUTTING UP A SLIDE WITH 

4542
03:32:19,019 --> 03:32:20,954
ALL THE HIT LIST, PEOPLE ALWAYS 

4543
03:32:20,954 --> 03:32:24,057
GO, OH, MY PROTEIN IS THERE.

4544
03:32:24,057 --> 03:32:27,027
AND THEN YOU GO, GREAT, CAN WE 

4545
03:32:27,027 --> 03:32:30,798
WORK ON THAT TOGETHER?

4546
03:32:30,798 --> 03:32:40,107
SO HERE WE ARE.

4547
03:32:40,107 --> 03:32:41,608
>> (NO MICROPHONE).

4548
03:32:41,608 --> 03:32:43,977
>> IT'S UBC-9, I THINK.

4549
03:32:43,977 --> 03:32:44,845
I CAN'T REMEMBER.

4550
03:32:44,845 --> 03:32:49,817
THEY'RE KNOWN, THE SUMO E1, E2, 

4551
03:32:49,817 --> 03:32:51,418
E3 ARE KNOWN FOR SUMOS.

4552
03:32:51,418 --> 03:32:52,820
I DON'T REMEMBER FOR SURE, 

4553
03:32:52,820 --> 03:32:53,053
THOUGH.

4554
03:32:53,053 --> 03:32:58,192
I'D HAVE TO THINK.

4555
03:32:58,192 --> 03:33:00,427
AND THE DRUGS ARE KNOWN WHAT 

4556
03:33:00,427 --> 03:33:02,129
THEY DO IN THAT CASCADE.

4557
03:33:02,129 --> 03:33:03,197
I THINK THEY DO IT EARLY IN THE 

4558
03:33:03,197 --> 03:33:13,240
CASCADE.

4559
03:33:13,240 --> 03:33:13,907
>> YOU'RE IN THE CENTER OF
THAT 

4560
03:33:13,907 --> 03:33:18,579
PICTURE TOO.

4561
03:33:18,579 --> 03:33:20,414
>> THESE ARE USUALLY TAKEN
AT 

4562
03:33:20,414 --> 03:33:20,681
BIRTHDAYS.

4563
03:33:20,681 --> 03:33:30,924
I DON'T KNOW.

4564
03:33:46,139 --> 03:33:47,608
>> THANK YOU, RECEIVE, FOR
A 

4565
03:33:47,608 --> 03:33:48,375
WONDERFUL TALK.

4566
03:33:48,375 --> 03:33:53,814
OUR NEXT SPEAKER NEEDS NO 

4567
03:33:53,814 --> 03:33:54,348
INTRODUCTION.

4568
03:33:54,348 --> 03:33:56,450
STEVE HUGHES HAS BEEN HERE IN 

4569
03:33:56,450 --> 03:33:59,620
FREDERICK ALMOST AS LONG AS I 

4570
03:33:59,620 --> 03:34:01,588
HAVE, AND LIKE ME, HE'S BEEN 

4571
03:34:01,588 --> 03:34:05,726
STUDYING RETROVIRUSES ALL THESE 

4572
03:34:05,726 --> 03:34:07,661
YEARS, AND I THINK AS TIME HAS 

4573
03:34:07,661 --> 03:34:10,297
GONE ON, WE'VE BECOME BETTER AND

4574
03:34:10,297 --> 03:34:13,834
BETTER FRIENDS.

4575
03:34:13,834 --> 03:34:14,801
I THINK I'LL LEAVE IT AT THAT.

4576
03:34:14,801 --> 03:34:24,912
STEVE?

4577
03:34:25,145 --> 03:34:26,813
>> I WAS TRYING TO THINK
BACK 

4578
03:34:26,813 --> 03:34:28,682
ABOUT WHEN I FIRST MET ALAN, AND

4579
03:34:28,682 --> 03:34:30,284
IT'S GOT TO HAVE BEEN MORE THAN 

4580
03:34:30,284 --> 03:34:32,352
FOUR DECADES AGO, AND IT MAY 

4581
03:34:32,352 --> 03:34:34,354
WELL HAVE BEEN IN COLD SPRING 

4582
03:34:34,354 --> 03:34:35,822
HARBOR, BUT I CERTAINLY MET HIM 

4583
03:34:35,822 --> 03:34:38,091
WHEN I CAME TO FREDERICK AND 

4584
03:34:38,091 --> 03:34:40,827
BEGAN TO REALLY BENEFIT FROM 

4585
03:34:40,827 --> 03:34:41,595
HAVING HIM AS A FRIEND AND A 

4586
03:34:41,595 --> 03:34:47,000
COLLEAGUE.

4587
03:34:47,000 --> 03:34:48,602
I'M GOING TO TELL A FEW STORIES 

4588
03:34:48,602 --> 03:34:52,172
OF ALAN'S, AND I HOPE TO DO 

4589
03:34:52,172 --> 03:34:54,174
REASONABLE JUSTICE TO THEM, THAT

4590
03:34:54,174 --> 03:34:55,242
I PARTICULARLY LIKE.

4591
03:34:55,242 --> 03:34:58,278
EACH ONE OF THOSE HAS TAUGHT ME 

4592
03:34:58,278 --> 03:34:58,545
SOMETHING.

4593
03:34:58,545 --> 03:35:01,915
I'VE TAKEN AWAY A GREAT DEAL OF 

4594
03:35:01,915 --> 03:35:02,716
POSITIVE FROM MY LONG 

4595
03:35:02,716 --> 03:35:06,119
ASSOCIATION WITH ALAN.

4596
03:35:06,119 --> 03:35:07,154
ONE OF THE INTERESTING THINGS OF

4597
03:35:07,154 --> 03:35:09,923
COURSE IS THAT THE IDEA OF 

4598
03:35:09,923 --> 03:35:12,059
ASSEMBLY, PUTTING THE VIRUS 

4599
03:35:12,059 --> 03:35:13,193
TOGETHER, ISN'T THE SAME 

4600
03:35:13,193 --> 03:35:14,695
NECESSARILY AS HAVING THE VIRUS 

4601
03:35:14,695 --> 03:35:16,363
READY TO BE INFECTIOUS.

4602
03:35:16,363 --> 03:35:18,865
YOU SOMEHOW HAVE TO CONVERT THE 

4603
03:35:18,865 --> 03:35:20,100
PROBLEM FROM PUTTING THINGS 

4604
03:35:20,100 --> 03:35:22,936
TOGETHER INTO BEING ABLE TO 

4605
03:35:22,936 --> 03:35:26,006
SOMETIMES TAKE THEM APART AGAIN.

4606
03:35:26,006 --> 03:35:28,175
FOR RETROVIRUSES AND OTHER 

4607
03:35:28,175 --> 03:35:30,644
VIRUSES AS WELL, THE PROPERLY 

4608
03:35:30,644 --> 03:35:37,951
ASSEMBLED POLYPROTEIN PRECU

4609
03:35:37,951 --> 03:35:41,989
PRECURSORS, THE IMMATURE VIRION 

4610
03:35:41,989 --> 03:35:44,324
NEED TO BE CONVERTED FROM 

4611
03:35:44,324 --> 03:35:46,960
PROTEASES INTO MATURE VIRION, 

4612
03:35:46,960 --> 03:35:48,695
AND THAT INVOLVES PROTEASES.

4613
03:35:48,695 --> 03:35:50,397
ONE OF THE BIZARRE THINGS ABOUT 

4614
03:35:50,397 --> 03:35:52,232
MLV IS THE MATURATION OF 

4615
03:35:52,232 --> 03:35:53,834
ENVELOPE -- AND THIS IS 

4616
03:35:53,834 --> 03:35:55,569
DIFFERENT FROM MLV THAN IT IS 

4617
03:35:55,569 --> 03:35:58,405
FOR THE OTHER VIRUSES -- IT'S A 

4618
03:35:58,405 --> 03:36:00,007
PRO-STAGE VIRUS AND IT INVOLVES 

4619
03:36:00,007 --> 03:36:01,575
TWO DIFFERENT PROTEASES.

4620
03:36:01,575 --> 03:36:02,943
THERE'S A HOST PROTEASE THAT 

4621
03:36:02,943 --> 03:36:05,178
SEPARATES THE SURFACE AND THE 

4622
03:36:05,178 --> 03:36:06,647
TRANSMEMORY PROTEIN, AND THAT 

4623
03:36:06,647 --> 03:36:08,548
CLEAVAGE IS NECESSARY, BUT IT'S 

4624
03:36:08,548 --> 03:36:12,419
NOT SUFFICIENT TO GIVE YOU 

4625
03:36:12,419 --> 03:36:13,186
FUSIGENIC ABILITY.

4626
03:36:13,186 --> 03:36:15,188
AND THE SECOND, AND THIS IS THE 

4627
03:36:15,188 --> 03:36:17,357
STORY OF ALAN'S THAT I LIKE SO 

4628
03:36:17,357 --> 03:36:19,793
MUCH, THERE'S A SECOND CLEAVAGE 

4629
03:36:19,793 --> 03:36:23,897
WHICH REMOVES ABOUT A 2 KB FROM 

4630
03:36:23,897 --> 03:36:26,333
THE END OF TRANSMEMORY THAT'S 

4631
03:36:26,333 --> 03:36:29,169
CARRIED OUT BY THE MLV PROTEASE.

4632
03:36:29,169 --> 03:36:32,305
AND IT TURNS OUT THIS IS A VERY 

4633
03:36:32,305 --> 03:36:34,374
IMPORTANT SAFETY FEATURE THAT 

4634
03:36:34,374 --> 03:36:36,977
PREVENTS THEM FROM BEING 

4635
03:36:36,977 --> 03:36:37,678
FUSIGENIC BEFORE THAT LITTLE 

4636
03:36:37,678 --> 03:36:39,413
PIECE IS REMOVED.

4637
03:36:39,413 --> 03:36:42,482
IN FACT, BECAUSE PROTEASE IS 

4638
03:36:42,482 --> 03:36:46,086
ACTIVATED WHEN THE VIRION IS 

4639
03:36:46,086 --> 03:36:47,621
RELEASES, THIS MEANS THE MLV ON 

4640
03:36:47,621 --> 03:36:50,590
THE SURFACE OF CELLS IS NOT 

4641
03:36:50,590 --> 03:36:51,625
FUSIGENIC, SO WHEN THAT PROTEIN 

4642
03:36:51,625 --> 03:36:54,394
IS EXPRESSED WITHOUT PROTEASE, 

4643
03:36:54,394 --> 03:36:58,565
THERE'S NO SYNCYTIA.

4644
03:36:58,565 --> 03:37:01,601
ALAN SHOWED THAT VERY ELEGANTLY,

4645
03:37:01,601 --> 03:37:03,036
AND I'LL SHOW YOU A SLIDE ON 

4646
03:37:03,036 --> 03:37:04,571
THIS IN JUST A MOMENT.

4647
03:37:04,571 --> 03:37:07,507
IF YOU EXPRESS MLV THAT LACKS 

4648
03:37:07,507 --> 03:37:11,678
THE P2E SAFETY PEPTIDE, THEN THE

4649
03:37:11,678 --> 03:37:14,648
PROTEIN IS FUSIGENIC ON THE 

4650
03:37:14,648 --> 03:37:17,784
SURFACE OF THE CELLS AND MAKES 

4651
03:37:17,784 --> 03:37:18,819
THE SYNCYTIA.

4652
03:37:18,819 --> 03:37:20,320
THESE ARE DESCRIBED IN TWO OF MY

4653
03:37:20,320 --> 03:37:21,154
FAVORITE ALAN PAPERS.

4654
03:37:21,154 --> 03:37:23,590
THE FIRST IS ACTUALLY THE 

4655
03:37:23,590 --> 03:37:25,058
ORIGINAL DEMONSTRATION THAT THE 

4656
03:37:25,058 --> 03:37:26,393
MLV PROTEASE IS INVOLVED, AND 

4657
03:37:26,393 --> 03:37:28,462
THE SECOND WHICH IS ACTUALLY THE

4658
03:37:28,462 --> 03:37:30,430
ONE THAT'S, I THINK, GENERALLY 

4659
03:37:30,430 --> 03:37:32,466
BETTER KNOWN, AND THIS WAS, I 

4660
03:37:32,466 --> 03:37:34,234
BELIEVE, MENTIONED BY ERIC, IS 

4661
03:37:34,234 --> 03:37:36,536
THE PAPER THAT SHOWS THAT THAT 

4662
03:37:36,536 --> 03:37:39,106
REMOVAL IS, IN FACT, REMOVING 

4663
03:37:39,106 --> 03:37:44,878
THE LITTLE SAFETY BIT.

4664
03:37:44,878 --> 03:37:46,713
SO THIS SLIDE THAT'S NEAREST ME,

4665
03:37:46,713 --> 03:37:49,216
THE SLIDE THAT'S ON THE RIGHT, 

4666
03:37:49,216 --> 03:37:53,754
MY RIGHT, SHOWED THAT THE 

4667
03:37:53,754 --> 03:37:55,922
ENVELOPE IF EXPRESSED WITHOUT 

4668
03:37:55,922 --> 03:37:56,957
PROTEASE, THE CELLS ARE 

4669
03:37:56,957 --> 03:37:58,792
PERFECTLY HAPPY, BUT IF YOU 

4670
03:37:58,792 --> 03:38:00,327
EXPRESS A DELETED PART OF THE 

4671
03:38:00,327 --> 03:38:02,395
PROTEIN THAT'S GOT THAT LITTLE 

4672
03:38:02,395 --> 03:38:05,866
BIT LOPPED OFF, THEN YOU MAKE 

4673
03:38:05,866 --> 03:38:06,967
LOTS OF SYNCYTIA.

4674
03:38:06,967 --> 03:38:09,169
THE MESSAGES HERE, THE MOST 

4675
03:38:09,169 --> 03:38:10,637
FUNDAMENTAL MESSAGES, THE ONE I 

4676
03:38:10,637 --> 03:38:12,672
LIKE THE BEST, IS THIS IS ONE OF

4677
03:38:12,672 --> 03:38:14,808
THE CLEAR DEMONSTRATION OF HOW, 

4678
03:38:14,808 --> 03:38:17,077
IN FACT, ELEGANTLY AND CAREFULLY

4679
03:38:17,077 --> 03:38:19,312
THE VIRUSES, EVEN REALLY WHAT WE

4680
03:38:19,312 --> 03:38:21,982
THINK OF AS VERY SIMPLE VIRUSES,

4681
03:38:21,982 --> 03:38:25,152
HOW CAREFULLY THEY MANAGE THEIR 

4682
03:38:25,152 --> 03:38:28,655
MATURATION AND HOW EXQUISITELY 

4683
03:38:28,655 --> 03:38:29,923
APPROPRIATE THEIR INTERACTIONS 

4684
03:38:29,923 --> 03:38:32,325
WITH THE HOST CELLS ARE.

4685
03:38:32,325 --> 03:38:35,662
IT TURNS OUT THAT OTHER 

4686
03:38:35,662 --> 03:38:36,897
INTERACTIONS, LIKE THE AVIAN 

4687
03:38:36,897 --> 03:38:39,266
VIRUSES I PRIMARILY WORKED WITH 

4688
03:38:39,266 --> 03:38:42,469
BEFORE HIV, AND HIV ITSELF, ALSO

4689
03:38:42,469 --> 03:38:46,139
REGULATE THE FUSIGENICITY OF 

4690
03:38:46,139 --> 03:38:48,408
THEIR ENVELOPE PROTEINS ON THE 

4691
03:38:48,408 --> 03:38:49,743
SURFACE OF THEIR PRODUCER CELLS.

4692
03:38:49,743 --> 03:38:53,113
BUT, IN FACT, THE UNDERLYING 

4693
03:38:53,113 --> 03:38:55,515
MECHANISMS BUT WHICH 

4694
03:38:55,515 --> 03:39:03,423
RETROVIRUSES CONTROL THIS USE 

4695
03:39:03,423 --> 03:39:04,624
PARTICULAR TRICKS.

4696
03:39:04,624 --> 03:39:06,693
THIS IS BEAUTIFUL TO HAVE PEOPLE

4697
03:39:06,693 --> 03:39:09,796
THAT LOVE NON-HIV RETROVIRUSES 

4698
03:39:09,796 --> 03:39:11,698
SPEAK BECAUSE ONE OF THE 

4699
03:39:11,698 --> 03:39:12,766
MESSAGES I THINK THIS SHOULD 

4700
03:39:12,766 --> 03:39:14,701
BRING HOME TO ALL OF THIS, AND 

4701
03:39:14,701 --> 03:39:15,969
ONE OF THE GREAT THEMES, I 

4702
03:39:15,969 --> 03:39:18,305
THINK, OF ALAN'S CAREER, IS IN 

4703
03:39:18,305 --> 03:39:19,172
SPITE OF THE FACT THAT PEOPLE 

4704
03:39:19,172 --> 03:39:21,575
THAT GIVE OUT THE MONEY DON'T 

4705
03:39:21,575 --> 03:39:24,177
SEEM TO THINK SO, STUDYING 

4706
03:39:24,177 --> 03:39:25,145
RETROVIRUSES OTHER THAN HIV IS 

4707
03:39:25,145 --> 03:39:27,480
STILL VERY VALUABLE.

4708
03:39:27,480 --> 03:39:29,182
AND ONE OF THE THINGS THAT IS 

4709
03:39:29,182 --> 03:39:32,352
SORT OF AN ASTOUNDING ASIDE IS 

4710
03:39:32,352 --> 03:39:35,922
THAT, IN FACT, ALTHOUGH WHILE 

4711
03:39:35,922 --> 03:39:39,259
HIV DOESN'T USE THE PROTOIATION 

4712
03:39:39,259 --> 03:39:41,494
AS A WAY TO REGULATE HOW FUSION 

4713
03:39:41,494 --> 03:39:42,929
IS CONTROLLED, THERE'S ACTUALLY 

4714
03:39:42,929 --> 03:39:48,301
A BEAUT OF HIV THAT WAS STUDIED 

4715
03:39:48,301 --> 03:39:51,137
BY ABDUL WAHIM WHEN HE WAS IN 

4716
03:39:51,137 --> 03:39:53,306
ERIC'S LAB, THAT DOES USE THIS 

4717
03:39:53,306 --> 03:39:53,573
MECHANISM.

4718
03:39:53,573 --> 03:39:54,975
I'M NOT GOING TO TRY TO SORT 

4719
03:39:54,975 --> 03:39:56,943
THROUGH THAT, BUT IF YOU'RE 

4720
03:39:56,943 --> 03:39:58,011
INTERESTED, YOU CAN ASK EITHER 

4721
03:39:58,011 --> 03:39:59,880
ABDUL OR ERIC, AND I WOULD 

4722
03:39:59,880 --> 03:40:01,114
RECOMMEND THAT YOU DO SO BECAUSE

4723
03:40:01,114 --> 03:40:03,516
IT'S A VERY CUTE STORY.

4724
03:40:03,516 --> 03:40:05,385
ONE OF THE STORIES -- AGAIN, ONE

4725
03:40:05,385 --> 03:40:08,188
OF MY -- THESE ARE ALL SOME OF 

4726
03:40:08,188 --> 03:40:12,058
MY FAVORITE ALAN STORIES, AND 

4727
03:40:12,058 --> 03:40:13,360
I'M NOT TRYING TO SUGGEST TO YOU

4728
03:40:13,360 --> 03:40:15,862
THAT I PICKED THE THINGS THAT IN

4729
03:40:15,862 --> 03:40:18,365
SOME COSMIC SENSE ARE THE MOST 

4730
03:40:18,365 --> 03:40:19,532
SCIENTIFICALLY IMPORTANT, 

4731
03:40:19,532 --> 03:40:21,568
ALTHOUGH THIS ONE, I'M GOING TO 

4732
03:40:21,568 --> 03:40:23,603
ARGUE, THAT'S ENORMOUS IMPACT, 

4733
03:40:23,603 --> 03:40:26,539
BUT THESE HAVE BEEN, FOR ME, 

4734
03:40:26,539 --> 03:40:32,846
VERY PERSONAL LY ENLIGHTENING.

4735
03:40:32,846 --> 03:40:34,447
AND EACH ONE OF THESE RESONATES 

4736
03:40:34,447 --> 03:40:36,249
WITH ME, AND I'LL TRY TO EXPLAIN

4737
03:40:36,249 --> 03:40:36,449
WHY.

4738
03:40:36,449 --> 03:40:37,784
THE ONE I'M GOING TO TALK ABOUT 

4739
03:40:37,784 --> 03:40:40,153
NOW IS THE DEMONSTRATION THAT 

4740
03:40:40,153 --> 03:40:47,661
THE MINOXIDIL PHOSPHATES, IP5 OR

4741
03:40:47,661 --> 03:40:48,895
IP6 ARE IMPORTANT IN THE 

4742
03:40:48,895 --> 03:40:52,299
ASSEMBLY, THE PROPER AND NORMAL 

4743
03:40:52,299 --> 03:40:54,968
ASSEMBLY OF HIV-GAG.

4744
03:40:54,968 --> 03:40:57,504
WE NOW KNOW THAT THE ROLE OF IP5

4745
03:40:57,504 --> 03:41:00,006
AND IP6 IS IMPORTANT NOT ONLY IN

4746
03:41:00,006 --> 03:41:02,142
THE APPROPRIATE ASSEMBLY OF GAG,

4747
03:41:02,142 --> 03:41:03,310
BUT IT'S ALSO SUBSEQUENTLY 

4748
03:41:03,310 --> 03:41:05,412
IMPORTANT IN THE PROPER ASSEMBLY

4749
03:41:05,412 --> 03:41:08,181
OF THE CAPSID PROTEIN TO FORM 

4750
03:41:08,181 --> 03:41:11,751
THE CAPSID WITHIN THE VIRION.

4751
03:41:11,751 --> 03:41:13,153
IT TURNS OUT -- AND I SUSPECT 

4752
03:41:13,153 --> 03:41:14,587
MOST OF THE PEOPLE IN THE 

4753
03:41:14,587 --> 03:41:15,989
AUDIENCE KNOW THIS -- THAT THE 

4754
03:41:15,989 --> 03:41:18,992
NEGATIVELY CHARGED PHOSPHATES OF

4755
03:41:18,992 --> 03:41:21,494
IP5 AND IP6 INTERACT WITH 

4756
03:41:21,494 --> 03:41:24,097
POSITIVELY CHARGED AMINO ACIDS, 

4757
03:41:24,097 --> 03:41:27,467
AND THEY STABILIZE HEXAMERIC 

4758
03:41:27,467 --> 03:41:30,470
FORMS OF EITHER GAG OR CAPSID.

4759
03:41:30,470 --> 03:41:32,472
THESE SUBSEQUENT RECOGNITIONS OF

4760
03:41:32,472 --> 03:41:36,509
THE IMPACT OF IP5 AND IP6 ON 

4761
03:41:36,509 --> 03:41:40,714
BOTH GAG ASSEMBLY AND CAPSID, 

4762
03:41:40,714 --> 03:41:42,682
AND EXACTLY HOW THEY DO IT -- 

4763
03:41:42,682 --> 03:41:43,650
AND I'LL SHOW YOU THE STRUCTURAL

4764
03:41:43,650 --> 03:41:45,151
INFORMATION IN A LIMITED WAY IN 

4765
03:41:45,151 --> 03:41:46,753
JUST A MOMENT -- ALL OF THESE 

4766
03:41:46,753 --> 03:41:48,822
SUBSEQUENT DISCOVERIES REST ON 

4767
03:41:48,822 --> 03:41:50,390
THE INITIAL DEMONSTRATION OF 

4768
03:41:50,390 --> 03:41:52,058
ALAN'S LABORATORY THAT THE IP5 

4769
03:41:52,058 --> 03:41:54,294
AND IP6 HAS AN IMPORTANT ROLE IN

4770
03:41:54,294 --> 03:42:02,635
THE ASSEMBLY OF GAG IN-VITRO.

4771
03:42:02,635 --> 03:42:03,603
YOU'VE ACTUALLY SEEN THIS 

4772
03:42:03,603 --> 03:42:05,138
REFERENCE BEFORE, AND THIS IS --

4773
03:42:05,138 --> 03:42:07,307
I WILL REMIND YOU AGAIN THIS IS 

4774
03:42:07,307 --> 03:42:09,409
NOT QUITE 25 YEARS AGO.

4775
03:42:09,409 --> 03:42:11,378
SO THIS IS A LONG WAY BACK, AND 

4776
03:42:11,378 --> 03:42:13,980
IT'S TAKEN ALL THIS TIME, I 

4777
03:42:13,980 --> 03:42:16,549
THINK, FOR PEOPLE TO PROPERLY 

4778
03:42:16,549 --> 03:42:23,590
APPRECIATE HOW IMPORTANT IP5 AND

4779
03:42:23,590 --> 03:42:25,025
IP6 ARE IN WHAT WE NOW KNOW 

4780
03:42:25,025 --> 03:42:27,160
ABOUT HIV, AND IT TURNS OUT 

4781
03:42:27,160 --> 03:42:28,428
SUBSEQUENTLY IN OTHER 

4782
03:42:28,428 --> 03:42:29,162
RETROVIRUSES AS WELL.

4783
03:42:29,162 --> 03:42:30,630
THIS IS A SLIGHTLY COBBLED 

4784
03:42:30,630 --> 03:42:31,931
TOGETHER SLIDE, AND I APOLOGIZE 

4785
03:42:31,931 --> 03:42:34,334
THAT IT'S NOT A BETTER ONE.

4786
03:42:34,334 --> 03:42:41,474
IT'S MY FAULT, NOT ALAN'S.

4787
03:42:41,474 --> 03:42:43,276
WHEN ALAN AND HIS COLLEAGUE, IN 

4788
03:42:43,276 --> 03:42:45,645
THIS PARTICULAR CASE, STEVE 

4789
03:42:45,645 --> 03:42:46,546
CAMPBELL, BEGAN TRYING TO 

4790
03:42:46,546 --> 03:42:48,581
ASSEMBLE GAG PARTICLES IN A TEST

4791
03:42:48,581 --> 03:42:49,949
TUBE, USING A SLIGHTLY MODIFIED 

4792
03:42:49,949 --> 03:42:51,451
FORM OF GAG, WHEN THEY FIRST 

4793
03:42:51,451 --> 03:42:53,520
STARTED DOING THAT, THE FIRST 

4794
03:42:53,520 --> 03:42:55,955
THING THEY SAW WAS THAT THEY 

4795
03:42:55,955 --> 03:43:00,593
COULD GET LITTLE VIRUS-LIKE 

4796
03:43:00,593 --> 03:43:02,662
PARTICLES, BUT THEY'RE VERY MUCH

4797
03:43:02,662 --> 03:43:06,833
THEIR OWN SIZE FOR THE 

4798
03:43:06,833 --> 03:43:07,167
EXPERIMENT.

4799
03:43:07,167 --> 03:43:08,635
THEY'RE TOO SMALL, CONSIDERABLY 

4800
03:43:08,635 --> 03:43:09,102
TOO SMALL.

4801
03:43:09,102 --> 03:43:11,571
THIS IS A BIT OF A CHEAT BECAUSE

4802
03:43:11,571 --> 03:43:13,206
THEY'RE SLIGHTLY COBBLED 

4803
03:43:13,206 --> 03:43:14,340
TOGETHER, SO I WON'T SWEAR TO 

4804
03:43:14,340 --> 03:43:15,975
YOU THE SIZE RATIOS ARE EXACTLY 

4805
03:43:15,975 --> 03:43:16,342
RIGHT.

4806
03:43:16,342 --> 03:43:17,610
BUT THE IDEA THAT THEY'RE WAY 

4807
03:43:17,610 --> 03:43:18,878
TOO SMALL IS CORRECT.

4808
03:43:18,878 --> 03:43:21,247
BUT THE THING THAT WAS 

4809
03:43:21,247 --> 03:43:31,658
ASTOUNDING WAS IF  IF 

4810
03:43:31,658 --> 03:43:33,693
RETICULOCYTE IS ADDED, YOU GET 

4811
03:43:33,693 --> 03:43:36,196
THE SIZE CORRECT, AND YOU KNOW 

4812
03:43:36,196 --> 03:43:37,664
THE SIZE IS CORRECT BECAUSE 

4813
03:43:37,664 --> 03:43:39,432
THERE'S HIV CORE FOR COMPARISON.

4814
03:43:39,432 --> 03:43:44,637
SOMETHING MAGICAL IN THE RABBIT 

4815
03:43:44,637 --> 03:43:45,805
RETICULOCYTE WAS ABLE TO CONVERT

4816
03:43:45,805 --> 03:43:47,073
THE WRONG ASSEMBLY INTO THE 

4817
03:43:47,073 --> 03:43:48,541
RIGHT ASSEMBLY, AND THEY SHOWED 

4818
03:43:48,541 --> 03:43:52,846
THAT MAGIC SOMETHING WAS IP5 AND

4819
03:43:52,846 --> 03:43:58,818
IP6, AND INSTEAD OF ADDING THE 

4820
03:43:58,818 --> 03:44:02,288
RETICULOCYTE, YOU ADD SIMPLY THE

4821
03:44:02,288 --> 03:44:04,023
APPROPRIATE CO-FACTOR.

4822
03:44:04,023 --> 03:44:05,458
THIS IS A DEMONSTRATION OF 

4823
03:44:05,458 --> 03:44:07,494
WHAT'S GOING ON TAKEN FROM A 

4824
03:44:07,494 --> 03:44:10,864
REVIEW THAT WAS WRITTEN BY ALAN.

4825
03:44:10,864 --> 03:44:13,433
SO YOU HAVE THE IMMATURE VIRION 

4826
03:44:13,433 --> 03:44:16,136
HERE AND YOU HAVE THE LITTLE 

4827
03:44:16,136 --> 03:44:18,471
PIECE THAT'S CAPSID.

4828
03:44:18,471 --> 03:44:22,442
OVER HERE YOU SEE THE C-TERMINAL

4829
03:44:22,442 --> 03:44:24,344
DOMAIN OF CAPSID EMBEDDED IN 

4830
03:44:24,344 --> 03:44:24,511
GAG.

4831
03:44:24,511 --> 03:44:26,746
IT'S GOT A LITTLE BIT OF A SPACE

4832
03:44:26,746 --> 03:44:28,381
FOR PEPTIDE 1 ATTACHED TO IT, 

4833
03:44:28,381 --> 03:44:34,888
AND IT'S A LITTLE HARD TO SEE.

4834
03:44:34,888 --> 03:44:37,524
BUT IP6 IS RIGHT THERE, AND THE 

4835
03:44:37,524 --> 03:44:39,692
PHOSPHATES OF IP6, IN THIS CASE,

4836
03:44:39,692 --> 03:44:44,531
ARE INTERACTING WITH A RING OF 

4837
03:44:44,531 --> 03:44:48,268
AMINO ACIDS HOLDING THE HEX 

4838
03:44:48,268 --> 03:44:52,138
TOGETHER, IN THIS CASE, 

4839
03:44:52,138 --> 03:44:52,705
HEXAMERIC GAG.

4840
03:44:52,705 --> 03:44:54,207
IN A SLIGHTLY DIFFERENT FASHION,

4841
03:44:54,207 --> 03:44:56,876
THE SAME SMALL MOLECULE IS ABLE 

4842
03:44:56,876 --> 03:45:01,147
TO BRING TOGETHER THE HEXAMERIC 

4843
03:45:01,147 --> 03:45:03,983
ARRAY OF CAPSID ITSELF.

4844
03:45:03,983 --> 03:45:05,518
THERE IT'S A LITTLE EASIER TO 

4845
03:45:05,518 --> 03:45:07,153
SEE, THE IP6 IN THE MIDDLE WITH 

4846
03:45:07,153 --> 03:45:09,756
THE PHOSPHATES AROUND THE EDGE 

4847
03:45:09,756 --> 03:45:12,425
AND THE 6 CAPSIDS.

4848
03:45:12,425 --> 03:45:15,028
IN THIS CASE, THIS IS THE MATURE

4849
03:45:15,028 --> 03:45:16,396
CAPSID IN THE MATURE VIRION.

4850
03:45:16,396 --> 03:45:18,198
AND ONE OF THE THINGS THAT'S 

4851
03:45:18,198 --> 03:45:20,733
INTERESTING ABOUT THIS TO ME IS,

4852
03:45:20,733 --> 03:45:24,304
IN FACT, ALTHOUGH THE SIMILAR 

4853
03:45:24,304 --> 03:45:26,706
TYPE OF INTERACTION IS TAKING 

4854
03:45:26,706 --> 03:45:28,508
PLACE IN BOTH CASES, IN WHICH 

4855
03:45:28,508 --> 03:45:32,011
YOU HAVE THE PHOSPHATES BEING A 

4856
03:45:32,011 --> 03:45:34,180
CHARGE, A RING OF CHARGE THAT 

4857
03:45:34,180 --> 03:45:37,450
BRINGS TOGETHER THE SIX 

4858
03:45:37,450 --> 03:45:38,551
COMPARTMENTS, WHETHER IT'S GAG 

4859
03:45:38,551 --> 03:45:41,354
OR CAPSID, EXACTLY WHERE THOSE 

4860
03:45:41,354 --> 03:45:43,223
ARE IN CAPSID IS DIFFERENT.

4861
03:45:43,223 --> 03:45:45,792
SO IF WE'RE TALKING ABOUT IN THE

4862
03:45:45,792 --> 03:45:49,963
CONTEXT OF GAG, THESE TWO 

4863
03:45:49,963 --> 03:45:53,266
LYSATES, ONE HERE AND ONE HERE 

4864
03:45:53,266 --> 03:46:02,775
RIGHT NEXT TO SB 1, AND YOU LOOK

4865
03:46:02,775 --> 03:46:05,845
AT THE MATURE GAG, IT'S IN A 

4866
03:46:05,845 --> 03:46:06,713
SLIGHTLY DIFFERENT PLACE AND 

4867
03:46:06,713 --> 03:46:07,981
DIFFERENT AMINO ACIDS.

4868
03:46:07,981 --> 03:46:09,716
IT'S THE SAME THEME, BUT IT'S 

4869
03:46:09,716 --> 03:46:11,284
MAKING USE OF THE INTERACTIONS, 

4870
03:46:11,284 --> 03:46:13,353
WHICH IS A SIMILAR INTERACTION, 

4871
03:46:13,353 --> 03:46:15,355
INVOLVING DIFFERENT AMINO ACIDS.

4872
03:46:15,355 --> 03:46:16,289
AND ONE OF THE THINGS THAT'S 

4873
03:46:16,289 --> 03:46:17,757
SORT OF CUTE ABOUT THAT, WHICH 

4874
03:46:17,757 --> 03:46:19,259
HAS ONLY RECENTLY BEEN 

4875
03:46:19,259 --> 03:46:21,060
RECOGNIZED, I THINK, IS THE 

4876
03:46:21,060 --> 03:46:23,129
INTERACTION WITH GAG THAT BRINGS

4877
03:46:23,129 --> 03:46:27,667
THE IP5 AND IP 6 INTO THE VIRION

4878
03:46:27,667 --> 03:46:31,504
WHERE IT CAN ACT TO NUCLEATE THE

4879
03:46:31,504 --> 03:46:32,705
ASSEMBLY OF THE CAPSID AT THE 

4880
03:46:32,705 --> 03:46:34,140
TIME AND PLACE THAT IT IS 

4881
03:46:34,140 --> 03:46:34,374
NEEDED.

4882
03:46:34,374 --> 03:46:36,142
OF COURSE, IN A SITUATION IN 

4883
03:46:36,142 --> 03:46:40,680
WHICH THE IP5 AND IP6 IS NOT 

4884
03:46:40,680 --> 03:46:45,852
BROUGHT IN WHICH THE INTERACTION

4885
03:46:45,852 --> 03:46:48,388
WITH GAG, IT WON'T BE THERE FOR 

4886
03:46:48,388 --> 03:46:53,926
CAPSID TO PROPERLY ASSEMBLE.

4887
03:46:53,926 --> 03:46:56,529
SO I THINK THE KEY IDEA HERE, AT

4888
03:46:56,529 --> 03:46:58,331
LEAST FOR ME, IS THAT THE 

4889
03:46:58,331 --> 03:47:02,368
DISCOVERY OF IP5 AND IP6, THE 

4890
03:47:02,368 --> 03:47:03,970
DISCOVERY OF THOSE TWO PROTEINS,

4891
03:47:03,970 --> 03:47:05,805
THOSE TWO SMALL MOLECULES, PLAY 

4892
03:47:05,805 --> 03:47:07,740
A CRUCIAL ROLE IN GAG ASSEMBLY, 

4893
03:47:07,740 --> 03:47:10,743
PAVED THE WAY FOR OUR CURRENT 

4894
03:47:10,743 --> 03:47:13,413
UNDERSTANDING MEMORY OF HOW GAG 

4895
03:47:13,413 --> 03:47:15,348
ASSEMBLES, BUT IN FACT HOW 

4896
03:47:15,348 --> 03:47:17,684
CAPSID ASSEMBLES, AND THOSE 

4897
03:47:17,684 --> 03:47:18,851
DISCOVERIES UNDERLIE OUR CURRENT

4898
03:47:18,851 --> 03:47:20,586
UNDERSTANDING OF HOW REVERSE 

4899
03:47:20,586 --> 03:47:23,623
TRANSCRIPTION TAKES PLACE IN 

4900
03:47:23,623 --> 03:47:24,324
CAPSIDS.

4901
03:47:24,324 --> 03:47:26,926
THAT ALSO HAS FACILITATED SOME 

4902
03:47:26,926 --> 03:47:27,894
REALLY IMPORTANT STRUCTURAL 

4903
03:47:27,894 --> 03:47:29,462
WORK, SOME OF WHICH YOU'VE SEEN 

4904
03:47:29,462 --> 03:47:31,464
FROM ME, OR MORE LIKELY IN TALKS

4905
03:47:31,464 --> 03:47:35,868
GIVEN BY OTHERS.

4906
03:47:35,868 --> 03:47:37,370
THE INTERESTING THING ABOUT THIS

4907
03:47:37,370 --> 03:47:38,538
FROM MY OWN PERSONAL PERSPECTIVE

4908
03:47:38,538 --> 03:47:40,340
IS THAT, WHEN THE GAG ASSEMBLY 

4909
03:47:40,340 --> 03:47:43,242
WORK WAS BEING DONE, ALAN'S LAB 

4910
03:47:43,242 --> 03:47:46,179
AND MY LAB HAD HELD JOINT LAB 

4911
03:47:46,179 --> 03:47:48,681
MEETI

4912
03:47:48,681 --> 03:47:49,949
MEETINGS, AND IT WAS STEVE 

4913
03:47:49,949 --> 03:47:52,151
CAMPBELL WHO DID THE ACTUAL WORK

4914
03:47:52,151 --> 03:47:55,355
ON THE ASSEMBLY OF GAG.

4915
03:47:55,355 --> 03:47:57,156
WHEN HE DISCOVERED THAT THERE 

4916
03:47:57,156 --> 03:48:07,700
WAS SOMETHING MAGIC IN RETIC

4917
03:48:11,471 --> 03:48:16,976
RETICULOSITE GLYSATES, HE WAS 

4918
03:48:16,976 --> 03:48:18,411
GOING TO IMPORT HIM.

4919
03:48:18,411 --> 03:48:21,080
I DECIDED TO KEEP MY BIG MOUTH 

4920
03:48:21,080 --> 03:48:23,750
SHUT, AND I DIDN'T TELL THEM HOW

4921
03:48:23,750 --> 03:48:25,551
HORRIBLE I THOUGHT HE WAS GOING 

4922
03:48:25,551 --> 03:48:28,721
TO BE TO PULL OUT THIS FACTOR 

4923
03:48:28,721 --> 03:48:38,030
FROM EITHER ONE.

4924
03:48:38,030 --> 03:48:38,865
I WAS QUITE CONFIDENT IT WAS 

4925
03:48:38,865 --> 03:48:41,401
GOING TO BE SOME BIG, LARGE, 

4926
03:48:41,401 --> 03:48:42,168
COMPLICATED PROTEIN THAT WAS 

4927
03:48:42,168 --> 03:48:44,537
GOING TO SOMEHOW MODIFY THE 

4928
03:48:44,537 --> 03:48:45,671
STRUCTURE OF THE GAG BITS, AND I

4929
03:48:45,671 --> 03:48:48,107
WAS JUST ALL WRONG.

4930
03:48:48,107 --> 03:48:51,944
THE OTHER THING THAT WAS 

4931
03:48:51,944 --> 03:48:53,679
IMPRESSIVE WAS IN VERY SHORT 

4932
03:48:53,679 --> 03:48:54,881
ORDER, THEY PULLED OUT THE 

4933
03:48:54,881 --> 03:48:56,115
MOLECULE AND IDENTIFIED IT.

4934
03:48:56,115 --> 03:48:57,850
THIS WAS A SEMINAL DISCOVERY 

4935
03:48:57,850 --> 03:48:59,218
THAT'S REALLY SET THE STAGE FOR 

4936
03:48:59,218 --> 03:49:01,888
A LOT OF WHAT'S GOING ON NOW 

4937
03:49:01,888 --> 03:49:03,589
BOTH IN TERMS OF CAPSID AND 

4938
03:49:03,589 --> 03:49:04,290
REVERSE TRANSCRIPTION AND A 

4939
03:49:04,290 --> 03:49:06,592
WHOLE BUNCH OF OTHER THINGS.

4940
03:49:06,592 --> 03:49:08,027
SO THIS IS ONE OF THE ONES I 

4941
03:49:08,027 --> 03:49:08,961
REALLY LIKE.

4942
03:49:08,961 --> 03:49:11,197
AND BOTH OF THE THINGS I'VE TOLD

4943
03:49:11,197 --> 03:49:13,633
YOU SO FAR IN THE LITTLE 

4944
03:49:13,633 --> 03:49:14,867
VIGNETTES ARE LITTLE VIGNETTES 

4945
03:49:14,867 --> 03:49:16,269
THAT OTHERS HAVE MENTIONED, BUT 

4946
03:49:16,269 --> 03:49:17,804
ONE OF THE ONES THAT ACTUALLY NO

4947
03:49:17,804 --> 03:49:19,005
ONE HAS TALKED ABOUT -- AND THIS

4948
03:49:19,005 --> 03:49:20,440
IS ACTUALLY A MEASURE OF THE 

4949
03:49:20,440 --> 03:49:21,607
FACT THAT THERE'S LOTS OF THINGS

4950
03:49:21,607 --> 03:49:22,975
TO TALK ABOUT WHEN WE'RE TALKING

4951
03:49:22,975 --> 03:49:26,345
ABOUT ALAN'S CAREER -- IS THAT, 

4952
03:49:26,345 --> 03:49:29,782
IN FACT, XMRV -- AND SOME OF YOU

4953
03:49:29,782 --> 03:49:31,017
MAY HAVE HAD EITHER GOOD OR BAD 

4954
03:49:31,017 --> 03:49:34,387
MEMORIES OF THIS.

4955
03:49:34,387 --> 03:49:35,988
XMRV IS A LABORATORY 

4956
03:49:35,988 --> 03:49:36,289
CONTAMINANT.

4957
03:49:36,289 --> 03:49:37,457
IT'S NOT A HUMAN PATHOGEN.

4958
03:49:37,457 --> 03:49:39,192
AS I SAID, SOME OF YOU MAY 

4959
03:49:39,192 --> 03:49:40,693
REMEMBER THE CLAIMS THAT ARE NOW

4960
03:49:40,693 --> 03:49:45,531
CLEARLY FALSE THAT A MURINE 

4961
03:49:45,531 --> 03:49:48,401
RETROVIRUS XMRV WAS FOUND IN 

4962
03:49:48,401 --> 03:49:50,369
PROSTATE TUMORS AND ASSOCIATED 

4963
03:49:50,369 --> 03:49:52,138
WITH CANCER AND ALL SORTS OF 

4964
03:49:52,138 --> 03:49:53,739
OTHER MAGIC THINGS DEPENDING ON 

4965
03:49:53,739 --> 03:49:54,941
WHAT DAY OF THE WEEK IT WAS.

4966
03:49:54,941 --> 03:49:57,944
I DON'T MEAN TO IMPLY THAT ALAN 

4967
03:49:57,944 --> 03:50:00,213
WAS THE ONLY LAB IN DRP TO WORK 

4968
03:50:00,213 --> 03:50:05,084
ON THIS, THERE WERE A NUMBER OF 

4969
03:50:05,084 --> 03:50:10,623
DRPs AND PIs THAT SORTED OUT ALL

4970
03:50:10,623 --> 03:50:11,958
THESE CLAIMS, BUT WHAT WAS 

4971
03:50:11,958 --> 03:50:13,025
REALLY IMPORTANT IS SOMEONE 

4972
03:50:13,025 --> 03:50:14,427
NEEDED TO LOOK CAREFULLY AT THE 

4973
03:50:14,427 --> 03:50:16,896
PROSTATE CANCER CLAIMS.

4974
03:50:16,896 --> 03:50:21,734
ALAN MADE USE OF HIS CONTACTS 

4975
03:50:21,734 --> 03:50:24,637
WITHIN THE NCI TO GET PROSTATE 

4976
03:50:24,637 --> 03:50:26,572
TUMOR SAMPLES, AND HE WAS VERY 

4977
03:50:26,572 --> 03:50:28,341
BOTHERED BY THE FACT THAT, WHEN 

4978
03:50:28,341 --> 03:50:31,077
THE PROSTATE SAMPLES WERE 

4979
03:50:31,077 --> 03:50:33,479
CAREFULLY ANALYZED BOTH BY PCR 

4980
03:50:33,479 --> 03:50:35,548
AND ANTIBODY STAINING, ALL THE 

4981
03:50:35,548 --> 03:50:36,883
CONTROLS WORKED AND ALL THE 

4982
03:50:36,883 --> 03:50:39,585
PROSTATE SAMPLES WERE NEGATIVE.

4983
03:50:39,585 --> 03:50:44,357
THIS IS THE MOST RELEVANT PAPER 

4984
03:50:44,357 --> 03:50:49,295
THAT DESCRIBES THAT.

4985
03:50:49,295 --> 03:50:50,863
THIS IS ONE OF THE PIECES OF 

4986
03:50:50,863 --> 03:50:52,498
INFORMATION, THIS IS AN ANTIBODY

4987
03:50:52,498 --> 03:50:53,733
STAIN, AND YOU CAN SEE THAT THE 

4988
03:50:53,733 --> 03:50:56,068
CONTROLS AT THE TOP HAVE THINGS 

4989
03:50:56,068 --> 03:50:57,403
THAT OBVIOUSLY STAIN STRONGLY 

4990
03:50:57,403 --> 03:51:01,741
WITH ANTIBODY THAT RECOGNIZES 

4991
03:51:01,741 --> 03:51:04,877
XMRV, AND SURE ENOUGH, BOTH THE 

4992
03:51:04,877 --> 03:51:06,946
NORMAL PROSTATE AND THE TERMINAL

4993
03:51:06,946 --> 03:51:08,414
SAMPLES ARE ALL NEGATIVE.

4994
03:51:08,414 --> 03:51:09,515
IT'S IMPORTANT TO STATE, THOUGH 

4995
03:51:09,515 --> 03:51:11,851
I'M NOT GOING TO SHOW ANY OF THE

4996
03:51:11,851 --> 03:51:14,520
DATA, THAT ALL OF THE DATA ALAN 

4997
03:51:14,520 --> 03:51:16,055
AND HIS COLLEAGUES OBTAINED 

4998
03:51:16,055 --> 03:51:19,458
USING PCR WAS EQUIVALENTLY 

4999
03:51:19,458 --> 03:51:20,259
NEGATIVE.

5000
03:51:20,259 --> 03:51:21,327
THERE WERE TWO THINGS THAT VERY 

5001
03:51:21,327 --> 03:51:22,795
MUCH IMPRESSED ME ABOUT THE WORK

5002
03:51:22,795 --> 03:51:24,397
THAT WAS DONE WITH THE PROSTATE 

5003
03:51:24,397 --> 03:51:26,132
SAMPLES IN ALAN'S LAB.

5004
03:51:26,132 --> 03:51:28,601
THE FIRST WAS, IN CONTRAST TO 

5005
03:51:28,601 --> 03:51:30,836
WHAT A NUMBER OF OTHER LABS, NOT

5006
03:51:30,836 --> 03:51:34,006
HERE IN FREDERICK BUT ELSEWHERE,

5007
03:51:34,006 --> 03:51:35,174
WERE DOING, ALL THE RESULTS WERE

5008
03:51:35,174 --> 03:51:37,577
NEGATIVE.

5009
03:51:37,577 --> 03:51:39,078
WHICH OF COURSE TURNS OUT TO BE 

5010
03:51:39,078 --> 03:51:40,246
THE RIGHT ANSWER.

5011
03:51:40,246 --> 03:51:42,582
THE SECOND THING THAT WAS 

5012
03:51:42,582 --> 03:51:45,851
IMPRESSIVE WAS HOW WORRIED ALAN 

5013
03:51:45,851 --> 03:51:47,954
WAS BECAUSE HE DIDN'T WANT TO 

5014
03:51:47,954 --> 03:51:49,021
TRY TO REACH A CONCLUSION THAT 

5015
03:51:49,021 --> 03:51:53,259
WAS BASED ON NEGATIVE DATA.

5016
03:51:53,259 --> 03:51:56,395
AND THAT, I THINK, HAS A REALLY 

5017
03:51:56,395 --> 03:51:58,431
IMPORTANT MESSAGE THAT RESONATES

5018
03:51:58,431 --> 03:52:00,199
VERY STRONGLY WITH ME, AND THAT 

5019
03:52:00,199 --> 03:52:02,168
IS YOU'VE GOT TO BE YOUR OWN 

5020
03:52:02,168 --> 03:52:03,836
HARSHEST CRITIC, AND YOU'VE GOT 

5021
03:52:03,836 --> 03:52:06,739
TO BE REALLY CAREFUL WITH 

5022
03:52:06,739 --> 03:52:11,210
NEGATIVE DATA.

5023
03:52:11,210 --> 03:52:12,778
ALAN, AFTER HAVING DONE HEROIC 

5024
03:52:12,778 --> 03:52:15,314
WORK ON THIS AND HAVING SPOKEN 

5025
03:52:15,314 --> 03:52:19,452
WITH HIS DRP COLLEAGUES, WAS 

5026
03:52:19,452 --> 03:52:20,486
CONVINCED, IN FACT, THAT THE 

5027
03:52:20,486 --> 03:52:23,889
ANSWER WAS NO, WHICH IT IS.

5028
03:52:23,889 --> 03:52:26,859
NOW, YOU JUST HEARD ABOUT ALAN'S

5029
03:52:26,859 --> 03:52:32,865
AFFECTION FOR GREAT MOVIES.

5030
03:52:32,865 --> 03:52:35,167
MY SESSIONS WITH HIM WAS MORE 

5031
03:52:35,167 --> 03:52:36,636
OVER BOOKS THAN MOVIES.

5032
03:52:36,636 --> 03:52:39,405
HE HAS A REAL PASSION FOR GREAT 

5033
03:52:39,405 --> 03:52:41,574
BOOKS, AND AGAIN AS OTHERS HAVE 

5034
03:52:41,574 --> 03:52:43,275
MENTIONED IN OUR DISCUSSIONS, I 

5035
03:52:43,275 --> 03:52:45,011
THINK I'VE GAINED MUCH MORE FROM

5036
03:52:45,011 --> 03:52:47,346
HIM IN TERMS OF THE BOOKS THAT 

5037
03:52:47,346 --> 03:52:50,049
WE'VE READ AND THE THINGS THAT 

5038
03:52:50,049 --> 03:52:51,851
WE'VE SUGGESTED THAN HE EVER 

5039
03:52:51,851 --> 03:52:52,485
GAINED FROM ME.

5040
03:52:52,485 --> 03:52:54,720
BUT I THINK HE HAD SOME 

5041
03:52:54,720 --> 03:52:56,756
PARTICULAR AFFECTION FOR I GUESS

5042
03:52:56,756 --> 03:52:59,158
WHAT I WOULD CALL RECENT SOCIAL 

5043
03:52:59,158 --> 03:52:59,392
HISTORY.

5044
03:52:59,392 --> 03:53:01,460
THESE ARE A SERIES OF THREE 

5045
03:53:01,460 --> 03:53:04,063
BOOKS BY TAYLOR BRANCH THAT ALAN

5046
03:53:04,063 --> 03:53:05,564
RECOMMENDED TO ME.

5047
03:53:05,564 --> 03:53:07,967
THEY RECOUNT THE HISTORY OF THE 

5048
03:53:07,967 --> 03:53:09,001
CIVIL RIGHTS MOVEMENT IN THE 

5049
03:53:09,001 --> 03:53:10,636
UNITED STATES THROUGH THE LIFE 

5050
03:53:10,636 --> 03:53:11,337
OF MARTIN LUTHER KING.

5051
03:53:11,337 --> 03:53:12,972
THEY'RE BY TAYLOR BRANCH.

5052
03:53:12,972 --> 03:53:15,007
THESE ARE SOME OF THE MOST 

5053
03:53:15,007 --> 03:53:19,345
MOVING BOOKS I'VE EVER READ, AND

5054
03:53:19,345 --> 03:53:20,613
THEY ARE SEMINAL, THEY ARE 

5055
03:53:20,613 --> 03:53:21,547
BEAUTIFULLY WRITTEN, AND THEY 

5056
03:53:21,547 --> 03:53:23,049
ARE COMPLETELY RELEVANT TODAY.

5057
03:53:23,049 --> 03:53:24,884
I WOULD RECOMMEND THEM TO EACH 

5058
03:53:24,884 --> 03:53:25,551
OF YOU.

5059
03:53:25,551 --> 03:53:26,686
IN FACT, I THINK THE COUNTRY 

5060
03:53:26,686 --> 03:53:28,587
WOULD BE A BETTER PLACE IF EVERY

5061
03:53:28,587 --> 03:53:30,656
MAN, WOMAN, AND CHILD OVER THE 

5062
03:53:30,656 --> 03:53:37,596
AGE OF ABOUT 12 WOULD READ THEM.

5063
03:53:37,596 --> 03:53:39,732
THEN I THINK YOU MAY HAVE SEEN 

5064
03:53:39,732 --> 03:53:41,167
ALAN IN AN ORIOLES CAP.

5065
03:53:41,167 --> 03:53:44,170
I WOULD TEND TO SAY THAT HIS 

5066
03:53:44,170 --> 03:53:45,538
ORIOLES CAPS TEND TO BE OF 

5067
03:53:45,538 --> 03:53:46,839
SLIGHTLY HIGHER MILEAGE THAN 

5068
03:53:46,839 --> 03:53:47,373
THIS ONE.

5069
03:53:47,373 --> 03:53:49,241
THE INTERESTING THING IS FROM 

5070
03:53:49,241 --> 03:53:50,743
TIME TO TIME I'VE BEEN STANDING 

5071
03:53:50,743 --> 03:53:53,245
NEAR ALAN OFTEN IN PLACES LIKE 

5072
03:53:53,245 --> 03:53:55,014
COLD SPRING HARBOR WHERE HE'S 

5073
03:53:55,014 --> 03:53:57,183
KNOWN, BUT PEOPLE DON'T KNOW HIM

5074
03:53:57,183 --> 03:53:58,818
PERSONALLY QUITE AS WELL AS I 

5075
03:53:58,818 --> 03:53:58,984
DO.

5076
03:53:58,984 --> 03:54:00,419
SOMEBODY WILL COME UP TO HIM, 

5077
03:54:00,419 --> 03:54:02,588
AND HE'S GOT AN ORIOLES BASEBALL

5078
03:54:02,588 --> 03:54:03,789
CAP, AND THEY'LL SAY, DO YOU 

5079
03:54:03,789 --> 03:54:04,890
LIKE THE ORIOLES?

5080
03:54:04,890 --> 03:54:07,193
AND ALAN WILL GET THAT BEAUTIFUL

5081
03:54:07,193 --> 03:54:08,728
SMILE, WHICH HE ACTUALLY HAS ON 

5082
03:54:08,728 --> 03:54:11,764
HIS FACE RIGHT NOW, AND HE'LL 

5083
03:54:11,764 --> 03:54:15,501
SAY, I LIKE THE BIRD.

5084
03:54:15,501 --> 03:54:19,638
I THINK THAT SUMS IT UP EXACTLY.

5085
03:54:19,638 --> 03:54:26,245
ALAN DOES LOVE BIRDS, AND THAT'S

5086
03:54:26,245 --> 03:54:27,513
BEEN MENTIONED, AND HE'S 

5087
03:54:27,513 --> 03:54:28,314
INCREDIBLY KNOWLEDGEABLE OF 

5088
03:54:28,314 --> 03:54:29,315
BIRDS IN GENERAL.

5089
03:54:29,315 --> 03:54:30,950
WALKING WITH HIM AND HAVING HIM 

5090
03:54:30,950 --> 03:54:32,384
TELL YOU WHICH BIRD IT IS 

5091
03:54:32,384 --> 03:54:33,385
LISTENING TO THE BIRD SONG IS AN

5092
03:54:33,385 --> 03:54:40,159
EDUCATION IN ITSELF.

5093
03:54:40,159 --> 03:54:41,994
IN SPITE OF SOME OF HIS BETTER 

5094
03:54:41,994 --> 03:54:43,496
EFFORTS, I LIKE BIRDS VERY MUCH,

5095
03:54:43,496 --> 03:54:44,597
BUT MY KNOWLEDGE OF BIRDS IS 

5096
03:54:44,597 --> 03:54:46,065
THEY'RE PRETTY AND I LIKE TO SEE

5097
03:54:46,065 --> 03:54:48,667
THEM FLY AND SOME OF THEM ARE 

5098
03:54:48,667 --> 03:54:48,934
TASTY.

5099
03:54:48,934 --> 03:54:51,737
SO I'LL FINISH BY TRYING TO PICK

5100
03:54:51,737 --> 03:54:54,507
A BIRD THAT HAS SOMETHING THAT 

5101
03:54:54,507 --> 03:54:55,808
BOTH HE AND I CAN LIKE.

5102
03:54:55,808 --> 03:55:00,379
THIS IS A BIRD THAT'S A 

5103
03:55:00,379 --> 03:55:01,013
FISHERMAN.

5104
03:55:01,013 --> 03:55:02,381
ALAN, IT'S BEEN A TREMENDOUS 

5105
03:55:02,381 --> 03:55:02,648
PLEASURE.

5106
03:55:02,648 --> 03:55:04,884
I'VE LEARNED A GREAT DEAL FROM 

5107
03:55:04,884 --> 03:55:09,522
OUR LONG ASSOCIATION, AND I HOPE

5108
03:55:09,522 --> 03:55:11,590
THAT LEARNING PROCESS IS NOT 

5109
03:55:11,590 --> 03:55:12,858
DONE, BY WHICH I MEAN, OF 

5110
03:55:12,858 --> 03:55:15,060
COURSE, THAT WE SHOULD STAY IN 

5111
03:55:15,060 --> 03:55:16,061
CONTACT SO WE CAN RECOMMEND 

5112
03:55:16,061 --> 03:55:17,429
OTHER GOOD THINGS FOR ME TO READ

5113
03:55:17,429 --> 03:55:18,497
AND HELP ME CONTINUE MY 

5114
03:55:18,497 --> 03:55:19,565
UNDERSTANDING OF VIRUSES AND HOW

5115
03:55:19,565 --> 03:55:19,832
THEY WORK.

5116
03:55:19,832 --> 03:55:30,142
THANK YOU SO MUCH.

5117
03:55:34,547 --> 03:55:35,815
I WILL ANSWER QUESTIONS IF 

5118
03:55:35,815 --> 03:55:37,082
ANYONE THINKS THEY HAVE THEM, 

5119
03:55:37,082 --> 03:55:38,818
BUT NOT ABOUT MLV.

5120
03:55:38,818 --> 03:55:41,787
>> I WANT TO SAY ABOUT
XMRV 

5121
03:55:41,787 --> 03:55:44,290
THAT, YES, I PRODUCED CLEARCUT 

5122
03:55:44,290 --> 03:55:46,725
NEGATIVE DATA, BUT THE ANSWER TO

5123
03:55:46,725 --> 03:55:51,597
THE PUZZLE CAME FROM JOHN AND 

5124
03:55:51,597 --> 03:55:51,797
RENEE.

5125
03:55:51,797 --> 03:55:54,533
THEY REALLY PUT IT TO REST IN A 

5126
03:55:54,533 --> 03:55:56,535
WAY THAT SIMPLY FINDING NEGATIVE

5127
03:55:56,535 --> 03:55:57,403
DATA COULD NOT.

5128
03:55:57,403 --> 03:56:03,909
>> AND I WOULD POINT OUT
THAT 

5129
03:56:03,909 --> 03:56:04,910
ALAN'S RESPONSE SAYS SOMETHING 

5130
03:56:04,910 --> 03:56:07,379
ELSE ABOUT HIM THAT I ALSO LIKE.

5131
03:56:07,379 --> 03:56:09,515
HE'S VERY GENEROUS WITH CREDIT 

5132
03:56:09,515 --> 03:56:12,124
FOR OTHERS. THANK YOU.

5133
03:56:12,124 --> 03:56:14,060
>> OUR NEXT SPEAKER
PROBABLY 

5134
03:56:14,060 --> 03:56:17,497
NEEDS NO INTRODUCTION.

5135
03:56:17,497 --> 03:56:19,265
IT'S JOHN COFFIN, WHO HAS BEEN 

5136
03:56:19,265 --> 03:56:22,101
AT THE FOREFRONT OF SO MANY 

5137
03:56:22,101 --> 03:56:28,441
DIFFERENT AVENUES OF RETROVIRUS 

5138
03:56:28,441 --> 03:56:29,542
RESEARCH THAT I WOULD NOT 

5139
03:56:29,542 --> 03:56:32,245
ATTEMPT TO TELL YOU MORE ABOUT 

5140
03:56:32,245 --> 03:56:32,879
IT.

5141
03:56:32,879 --> 03:56:35,314
JUST ALWAYS BRINGS A WONDERFUL, 

5142
03:56:35,314 --> 03:56:44,323
ENLIGHTENED PERSPECTIVE.

5143
03:56:44,323 --> 03:56:47,260
HE ALWAYS INCLUDES A WONDERFUL 

5144
03:56:47,260 --> 03:56:48,294
EVOLUTION IN WHATEVER HE'S 

5145
03:56:48,294 --> 03:56:49,495
THINKING ABOUT, AND I HAVE 

5146
03:56:49,495 --> 03:56:50,930
CERTAINLY LEARNED MORE THAN I 

5147
03:56:50,930 --> 03:56:52,498
COULD SAY FROM MY LONG 

5148
03:56:52,498 --> 03:56:53,599
ASSOCIATION WITH JOHN.

5149
03:56:53,599 --> 03:56:54,834
SO I THINK I'LL JUST LEAVE IT 

5150
03:56:54,834 --> 03:56:55,668
WITH THAT.

5151
03:56:55,668 --> 03:57:05,778
JOHN?

5152
03:57:11,050 --> 03:57:12,518
>> THANK YOU, ALAN.

5153
03:57:12,518 --> 03:57:13,586
THIS HAS BEEN A WONDERFUL EVENT,

5154
03:57:13,586 --> 03:57:14,554
I HAVE TO SAY.

5155
03:57:14,554 --> 03:57:16,689
THE STORY I'M ABOUT TO TELL YOU,

5156
03:57:16,689 --> 03:57:18,591
I HOPE WILL SOMEWHAT CLOSE THE 

5157
03:57:18,591 --> 03:57:20,726
CIRCLE BETWEEN WHAT JONATHAN 

5158
03:57:20,726 --> 03:57:27,033
SAID EARLIER TODAY AND GOING ON 

5159
03:57:27,033 --> 03:57:29,001
TO A DIFFERENT MODEL.

5160
03:57:29,001 --> 03:57:31,504
BEFORE I DO THAT -- AND SOME OF 

5161
03:57:31,504 --> 03:57:32,838
YOU MAY RECOGNIZE THESE TWO GUYS

5162
03:57:32,838 --> 03:57:33,205
HERE.

5163
03:57:33,205 --> 03:57:34,206
SOME OF THE OLDER ONES AMONG 

5164
03:57:34,206 --> 03:57:39,045
YOU.

5165
03:57:39,045 --> 03:57:41,047
BEFORE I DO THAT, I'VE GOT TO 

5166
03:57:41,047 --> 03:57:51,591
FIGURE OUT HOW TO ADVANCE HERE.

5167
03:57:55,628 --> 03:57:58,230
WHEN I FIRST MET ALAN, 

5168
03:57:58,230 --> 03:57:59,498
PRESUMABLY AT A COLD SPRING 

5169
03:57:59,498 --> 03:58:01,867
HARBOR MEETING, AS THEY WERE 

5170
03:58:01,867 --> 03:58:08,874
CALLED AT THE TIME, TUMOR VIRUS 

5171
03:58:08,874 --> 03:58:09,508
MEETING.

5172
03:58:09,508 --> 03:58:11,811
BEFORE THAT, WE WERE ACTUALLY 

5173
03:58:11,811 --> 03:58:13,846
CUSTOMER INS IN AN INTERESTING 

5174
03:58:13,846 --> 03:58:16,882
WAY, IS THAT IS THAT OUR THESIS 

5175
03:58:16,882 --> 03:58:21,220
MENTORS WERE RESPECTIVELY HARRY 

5176
03:58:21,220 --> 03:58:24,724
RUBIN AND HOWARD TENEMAN.

5177
03:58:24,724 --> 03:58:27,493
THOSE OF YOU WHO KNOW ABOUT THE 

5178
03:58:27,493 --> 03:58:33,332
HISTORY OF THIS, HARRY AND 

5179
03:58:33,332 --> 03:58:34,500
HOWARD WERE TOGETHER IN A LAB, 

5180
03:58:34,500 --> 03:58:37,136
AND THEY DID THE SEMINAL 

5181
03:58:37,136 --> 03:58:39,005
EXPERIMENT THAT ACTUALLY 

5182
03:58:39,005 --> 03:58:42,475
REFERRED HOWARD TO DETERMINE 

5183
03:58:42,475 --> 03:58:45,511
RETROVIRUS MUST REPLICATE BY A 

5184
03:58:45,511 --> 03:58:47,246
STABLED CELLULAR INTERMEDIATE, 

5185
03:58:47,246 --> 03:58:51,150
WHICH HE REFERRED TO AS THE PRO 

5186
03:58:51,150 --> 03:58:51,550
VIRUS.

5187
03:58:51,550 --> 03:58:53,519
THEN HE INFERRED FROM THAT THE 

5188
03:58:53,519 --> 03:58:55,688
PRO VIRUS HAD TO BE DNA, EVEN 

5189
03:58:55,688 --> 03:58:56,622
THOUGH IT WAS WELL-KNOWN ALREADY

5190
03:58:56,622 --> 03:58:58,691
EVEN AT THE TIME, THIS WAS LATE 

5191
03:58:58,691 --> 03:59:01,627
'50s AND EARLY '60s, THAT THE 

5192
03:59:01,627 --> 03:59:02,862
VIRUS GENOME WAS RNA.

5193
03:59:02,862 --> 03:59:04,697
THERE WAS NO DNA IN THE VIRUS.

5194
03:59:04,697 --> 03:59:05,231
IT HAD ALREADY BEEN WELL 

5195
03:59:05,231 --> 03:59:09,835
ESTABLISHED.

5196
03:59:09,835 --> 03:59:11,904
BUT THAT KIND OF INFERENCE DID 

5197
03:59:11,904 --> 03:59:14,473
NOT GO SIT WELL WITH HARRY, WHO 

5198
03:59:14,473 --> 03:59:16,609
WAS NOT AN INFERENTIAL 

5199
03:59:16,609 --> 03:59:16,876
SCIENTIST.

5200
03:59:16,876 --> 03:59:21,580
HE HAD TO SEE SOLID DATA 

5201
03:59:21,580 --> 03:59:23,149
INCONTROVERTIBLE BEFORE HE WOULD

5202
03:59:23,149 --> 03:59:23,783
BELIEVE ANYTHING.

5203
03:59:23,783 --> 03:59:25,284
THEY HAD A FAMOUS FEUD ABOUT 

5204
03:59:25,284 --> 03:59:27,920
THIS FOR QUITE A NUMBER OF YEARS

5205
03:59:27,920 --> 03:59:29,989
UNTIL -- THAT WAS UNTIL THERE 

5206
03:59:29,989 --> 03:59:33,459
WAS IN 1970, THERE WAS A CANCER 

5207
03:59:33,459 --> 03:59:37,396
CONGRESS IN HOUSTON, IF I 

5208
03:59:37,396 --> 03:59:39,699
REMEMBER CORRECTLY, AND HOWARD 

5209
03:59:39,699 --> 03:59:42,868
PRESEN

5210
03:59:42,868 --> 03:59:45,671
PRESENTED THE DISCOVERY REVERSE 

5211
03:59:45,671 --> 03:59:46,906
TRANSCRIPTICS AND ALL THE 

5212
03:59:46,906 --> 03:59:48,474
EVIDENCE FOR THAT IN A SESSION 

5213
03:59:48,474 --> 03:59:50,876
AT THE MEETING WHERE HARRY WAS 

5214
03:59:50,876 --> 03:59:51,977
THE CHAIR.

5215
03:59:51,977 --> 03:59:54,513
I REMEMBER HIM COMING BACK ALL 

5216
03:59:54,513 --> 03:59:55,614
GLOWING BECAUSE HARRY KEPT 

5217
03:59:55,614 --> 03:59:57,650
SAYING, I DON'T BELIEVE IT, I 

5218
03:59:57,650 --> 03:59:58,751
DON'T BELIEVE IT, AND HOWARD 

5219
03:59:58,751 --> 04:00:00,519
WOULD SAY THIS IS THE DATA, AND 

5220
04:00:00,519 --> 04:00:03,155
SURE ENOUGH, IT WAS THE DATA.

5221
04:00:03,155 --> 04:00:04,557
SURE ENOUGH, THE REST IS 

5222
04:00:04,557 --> 04:00:05,024
HISTORY.

5223
04:00:05,024 --> 04:00:06,926
IN ANY CASE, THEY NEVER DID 

5224
04:00:06,926 --> 04:00:07,660
SPEAK TO EACH OTHER AGAIN AS FAR

5225
04:00:07,660 --> 04:00:17,837
AS I KNOW.

5226
04:00:19,105 --> 04:00:20,172
>> HARRY LEFT THE FIELD.

5227
04:00:20,172 --> 04:00:22,108
>> EXACTLY.

5228
04:00:22,108 --> 04:00:24,210
HE GOT MUCH MORE INTO CELL 

5229
04:00:24,210 --> 04:00:26,212
BIOLOGY KINDS OF THINGS.

5230
04:00:26,212 --> 04:00:32,051
BUT ALAN AT COLD SPRING HARBOR 

5231
04:00:32,051 --> 04:00:34,720
WAS HARDLY ALAN'S FIRST COLD 

5232
04:00:34,720 --> 04:00:36,555
SPRING HARBOR EXPERIENCE, AND I 

5233
04:00:36,555 --> 04:00:38,090
HAD THE GOOD FORTUNE OF BEING 

5234
04:00:38,090 --> 04:00:40,459
ABLE TO CONTACT THE COLD SPRING 

5235
04:00:40,459 --> 04:00:41,727
HARBOR PHOTOGRAPHER, WHO WE ALL 

5236
04:00:41,727 --> 04:00:44,396
KNOW ACTUALLY FROM THE MEETING, 

5237
04:00:44,396 --> 04:00:47,733
AND THE ARCHIVIST WHO MAINTAINS 

5238
04:00:47,733 --> 04:00:51,070
A VERY GOOD ARCHIVE OF PAST 

5239
04:00:51,070 --> 04:00:53,239
MEETINGS, AND ASKED THEM TO FIND

5240
04:00:53,239 --> 04:00:55,508
ME PICTURES OF ALAN REIN, AND 

5241
04:00:55,508 --> 04:00:57,276
THEY SAID, WELL, WE'VE GOT ONE 

5242
04:00:57,276 --> 04:01:01,280
FROM 1961.

5243
04:01:01,280 --> 04:01:02,815
I DON'T KNOW IF ALAN HIMSELF HAS

5244
04:01:02,815 --> 04:01:04,850
A COPY OF THIS.

5245
04:01:04,850 --> 04:01:09,288
SO THAT WAS ALAN, AS I THINK 

5246
04:01:09,288 --> 04:01:10,823
ABOUT A SENIOR IN COLLEGE DOING 

5247
04:01:10,823 --> 04:01:13,926
A PROJECT AT COLD SPRING HARBOR.

5248
04:01:13,926 --> 04:01:15,995
AS YOU CAN SEE FROM THE 

5249
04:01:15,995 --> 04:01:17,062
PARAPHERNALIA AROUND THERE, IT 

5250
04:01:17,062 --> 04:01:18,764
WAS A FLY PROJECT HE WAS WORKING

5251
04:01:18,764 --> 04:01:18,964
ON.

5252
04:01:18,964 --> 04:01:20,332
HE COULD TELL YOU ALL ABOUT IT 

5253
04:01:20,332 --> 04:01:30,676
PROBABLY IF YOU ASK.

5254
04:01:30,943 --> 04:01:32,645
ACTUALLY, I GOT ABOUT 60 OR SO 

5255
04:01:32,645 --> 04:01:34,880
PICTURES FROM THEM, WHICH I 

5256
04:01:34,880 --> 04:01:37,716
CALLED OUT A FEW TO SHARE WITH 

5257
04:01:37,716 --> 04:01:37,917
YOU.

5258
04:01:37,917 --> 04:01:39,351
THIS ACTUALLY WAS A MEETING THAT

5259
04:01:39,351 --> 04:01:42,254
I ORGANIZED, ALONG WITH IRVIN 

5260
04:01:42,254 --> 04:01:44,223
CHEN IN THE MIDDLE, AND THIS WAS

5261
04:01:44,223 --> 04:01:45,124
A PICTURE OF THE SESSION CHAIRS 

5262
04:01:45,124 --> 04:01:47,393
AT THE MEETING, AND ALAN IS IN 

5263
04:01:47,393 --> 04:01:48,627
THE BACTERIO.

5264
04:01:48,627 --> 04:01:49,829
TWO OF THE PEOPLE YOU MIGHT 

5265
04:01:49,829 --> 04:01:50,896
RECOGNIZE, NANCY HOPKINS OVER 

5266
04:01:50,896 --> 04:01:53,032
THERE ON THE RIGHT THERE.

5267
04:01:53,032 --> 04:01:55,768
BY THE WAY, WHO WILL BE GIVING A

5268
04:01:55,768 --> 04:01:58,170
TALK IN A COUPLE OF WEEKS -- I 

5269
04:01:58,170 --> 04:02:01,106
THINK IN BETHESDA ACTUALLY -- 

5270
04:02:01,106 --> 04:02:03,309
BUT IT'S VERY WORTH TUNING INTO 

5271
04:02:03,309 --> 04:02:05,544
TO HEAR WHAT SHE HAS TO SAY.

5272
04:02:05,544 --> 04:02:07,112
I'VE HEARD IT BEFORE.

5273
04:02:07,112 --> 04:02:08,914
DAVID HO IS THERE, A NUMBER OF 

5274
04:02:08,914 --> 04:02:12,751
PEOPLE YOU KNOW, DON BLAIR, 

5275
04:02:12,751 --> 04:02:15,254
KAREN IS THERE.

5276
04:02:15,254 --> 04:02:16,689
MUCH YOUNGER, THIS WAS 1991, 

5277
04:02:16,689 --> 04:02:20,993
THIS MEETING.

5278
04:02:20,993 --> 04:02:25,030
AND HERE'S ANOTHER MEETING.

5279
04:02:25,030 --> 04:02:35,507
HERE'S ALAN WITH TAYE ZHANG.

5280
04:02:40,746 --> 04:02:41,881
OH, YOU'RE ABSOLUTELY RIGHT.

5281
04:02:41,881 --> 04:02:44,350
THIS WAS SAID TO ME AS BEING 

5282
04:02:44,350 --> 04:02:45,684
ALAN, BUT YOU'RE ABSOLUTELY 

5283
04:02:45,684 --> 04:02:45,885
RIGHT.

5284
04:02:45,885 --> 04:02:49,955
THAT'S ALAN.

5285
04:02:49,955 --> 04:02:52,458
AND WITH DAVID BALTIMORE AND ANN

5286
04:02:52,458 --> 04:02:54,326
SKULKA AT THE 40th ANNIVERSARY 

5287
04:02:54,326 --> 04:02:56,795
OF REVERSE TRANSCRIPTICS, SO IT 

5288
04:02:56,795 --> 04:02:58,697
WOULD HAVE BEEN 2010.

5289
04:02:58,697 --> 04:03:00,733
AND HERE HE IS AT POSTER 

5290
04:03:00,733 --> 04:03:02,801
SESSIONS AT COLD SPRING HARBOR 

5291
04:03:02,801 --> 04:03:04,536
LOOKING EITHER SKEPTICAL, AS HE 

5292
04:03:04,536 --> 04:03:06,505
SOMETIMES DOES, OR ENJOYING A 

5293
04:03:06,505 --> 04:03:07,006
GOOD JOKE.

5294
04:03:07,006 --> 04:03:09,341
THEN PROBABLY IN THE MIDDLE OF 

5295
04:03:09,341 --> 04:03:11,043
THE ROOM OUTSIDE AWAY FROM THE 

5296
04:03:11,043 --> 04:03:11,744
POSTERS.

5297
04:03:11,744 --> 04:03:13,712
AND FINALLY, ALAN ALWAYS DOES 

5298
04:03:13,712 --> 04:03:15,447
TEND TO STAND OUT IN A CROWD A 

5299
04:03:15,447 --> 04:03:15,948
LITTLE BIT.

5300
04:03:15,948 --> 04:03:18,150
YOU'RE VERY EASY TO SPOT IN THIS

5301
04:03:18,150 --> 04:03:19,885
PICTURE, HARDER TO SPOT ARE THE 

5302
04:03:19,885 --> 04:03:22,488
THREE GUYS IN THE FRONT THERE 

5303
04:03:22,488 --> 04:03:27,393
THAT YOU MIGHT RECOGNIZE.

5304
04:03:27,393 --> 04:03:31,096
>> THIS IS THE ONE WITH
THE 

5305
04:03:31,096 --> 04:03:36,902
SILVER HAIR?

5306
04:03:36,902 --> 04:03:43,409
>> THAT'S YOUNG BURTOWSKY,
A 

5307
04:03:43,409 --> 04:03:45,311
STAPLE AT COLD SPRING HARBOR.

5308
04:03:45,311 --> 04:03:47,179
AT THE COCKTAIL PARTY, WE ALWAYS

5309
04:03:47,179 --> 04:03:48,847
TALK TO HIM ACTUALLY.

5310
04:03:48,847 --> 04:03:51,417
AND THIS IS THANKS TO THE COLD 

5311
04:03:51,417 --> 04:03:52,718
SPRING HARBOR LABORATORY PEOPLE 

5312
04:03:52,718 --> 04:03:54,420
WHO PROVIDED THESE PICTURES.

5313
04:03:54,420 --> 04:03:56,288
THEY HAVE A MARVELOUS COLLECTION

5314
04:03:56,288 --> 04:03:57,756
OF PICTURES FROM THESE MEETINGS.

5315
04:03:57,756 --> 04:03:59,425
BACK TO THE STORY.

5316
04:03:59,425 --> 04:04:02,361
WE ALL KNOW NOW ABOUT ENDOGENOUS

5317
04:04:02,361 --> 04:04:03,762
RETROVIRUSES, AND JONATHAN GAVE 

5318
04:04:03,762 --> 04:04:06,699
A LOVELY INTRODUCTION AT THE 

5319
04:04:06,699 --> 04:04:06,966
BEGINNING.

5320
04:04:06,966 --> 04:04:09,535
THIS IS STILL NOW A VERY LARGE 

5321
04:04:09,535 --> 04:04:10,669
AREA OF STUDY NOW.

5322
04:04:10,669 --> 04:04:14,473
WHAT THESE VIRUSES HAVE DONE IN 

5323
04:04:14,473 --> 04:04:18,110
TERMS OF THEIR EFFECTS ON THE 

5324
04:04:18,110 --> 04:04:19,378
EVOLUTION AND GENE EXPRESSION 

5325
04:04:19,378 --> 04:04:21,880
AND ALL KINDS OF OTHER ASPECTS 

5326
04:04:21,880 --> 04:04:28,787
OF US BEING US, THERE IS, AS I 

5327
04:04:28,787 --> 04:04:31,190
SAY, ONE OF THE REMARKABLE 

5328
04:04:31,190 --> 04:04:33,525
FINDINGS OF THE HUMAN GENOME 

5329
04:04:33,525 --> 04:04:35,127
PROJECT IS THERE'S MORE VIRUSES 

5330
04:04:35,127 --> 04:04:37,363
IN US THAN US IN US.

5331
04:04:37,363 --> 04:04:40,132
WE HAVE SOMETHING LIKE 80,000 

5332
04:04:40,132 --> 04:04:42,868
ENDOGENOUS PRO VIRUSES COMPARED 

5333
04:04:42,868 --> 04:04:44,403
TO LIKE 20,000 GENES.

5334
04:04:44,403 --> 04:04:45,971
THESE ARE KNOWN TO DO LOTS OF 

5335
04:04:45,971 --> 04:04:50,075
THINGS, LIKE BRINGING IN NEW 

5336
04:04:50,075 --> 04:04:53,312
GENES, MODIFYING GENE 

5337
04:04:53,312 --> 04:04:53,879
EXPRESSION.

5338
04:04:53,879 --> 04:04:55,748
THEY CAN BECOME PARTS OF A LONG 

5339
04:04:55,748 --> 04:04:58,684
LINE OF CODING RNAs AND THINGS 

5340
04:04:58,684 --> 04:05:01,820
LIKE THAT.

5341
04:05:01,820 --> 04:05:07,993
IN CASE OF MICE THEY CAN 

5342
04:05:07,993 --> 04:05:09,028
REPLICATE AND CAUSE DISEASE, 

5343
04:05:09,028 --> 04:05:10,696
WHICH IS NOT TRUE IN HUMANS, 

5344
04:05:10,696 --> 04:05:11,830
DESPITE THE EFFORTS OF MANY 

5345
04:05:11,830 --> 04:05:13,432
PEOPLE TO TRY TO SHOW THAT IT 

5346
04:05:13,432 --> 04:05:13,599
IS.

5347
04:05:13,599 --> 04:05:15,067
AND TO OUR INTEREST, THEY 

5348
04:05:15,067 --> 04:05:18,270
PROVIDE A FOSSIL RECORD OF THE 

5349
04:05:18,270 --> 04:05:19,204
HOST-VIRUS INTERACTION GOING 

5350
04:05:19,204 --> 04:05:21,340
BACK MANY MILLIONS OF YEARS.

5351
04:05:21,340 --> 04:05:22,441
JONATHAN WAS TALKING ABOUT 47 

5352
04:05:22,441 --> 04:05:24,543
MILLION, I THINK, IN HIS.

5353
04:05:24,543 --> 04:05:25,744
WE DON'T BACK QUITE THAT FAR 

5354
04:05:25,744 --> 04:05:27,413
WITH THE ONES WE'RE LOOKING AT, 

5355
04:05:27,413 --> 04:05:29,114
BUT WE GO BACK A LONG WAYS.

5356
04:05:29,114 --> 04:05:32,718
AND I THINK THAT ONE OF THE MOST

5357
04:05:32,718 --> 04:05:38,924
OVERLOOKED THINGS IS BECOMING, 

5358
04:05:38,924 --> 04:05:41,193
THANKS TO JONATHAN AND THE WORK 

5359
04:05:41,193 --> 04:05:43,996
OF MANY OTHERS, UNDERSTAND IS 

5360
04:05:43,996 --> 04:05:46,265
PROTECTION FROM INFECTION BY 

5361
04:05:46,265 --> 04:05:49,735
RELATED EXOGENOUS VIRUSES.

5362
04:05:49,735 --> 04:05:53,305
I WOULD ARGUE THAT, PROBABLY THE

5363
04:05:53,305 --> 04:05:55,674
REASON THESE VIRUSES HAVE GONE 

5364
04:05:55,674 --> 04:05:58,077
EXTINCT IS LARGELY DUE TO THIS 

5365
04:05:58,077 --> 04:05:58,811
HAPPENING.

5366
04:05:58,811 --> 04:06:03,382
SOONER OR LATER A PRO VIRUS GETS

5367
04:06:03,382 --> 04:06:05,017
INTEGRATED INTO THE GERMLINE OF 

5368
04:06:05,017 --> 04:06:07,519
THE HOST, AND IT BLOCKS THE 

5369
04:06:07,519 --> 04:06:09,455
INTERACTION OF THE INCOMING 

5370
04:06:09,455 --> 04:06:10,856
VIRUS IN SOME IMPORTANT HOST 

5371
04:06:10,856 --> 04:06:13,192
CELL MACHINERY OR DOES SOMETHING

5372
04:06:13,192 --> 04:06:15,594
ELSE OF INTEREST THAT PREVENTS 

5373
04:06:15,594 --> 04:06:17,463
INJECTION OF THAT HOST BY THE 

5374
04:06:17,463 --> 04:06:18,130
SAME VIRUS FROM OUTSIDE.

5375
04:06:18,130 --> 04:06:20,466
IN FACT, IN LOOKING AT MLV SOME 

5376
04:06:20,466 --> 04:06:21,900
YEARS AGO, WE PUBLISHED THAT YOU

5377
04:06:21,900 --> 04:06:23,402
COULD ACTUALLY SEE THIS BACK AND

5378
04:06:23,402 --> 04:06:26,538
FORTH AT THE LEVEL RECEPTOR 

5379
04:06:26,538 --> 04:06:28,474
REACTION, LOOKING AT ENDOGENOUS 

5380
04:06:28,474 --> 04:06:29,241
MLVs.

5381
04:06:29,241 --> 04:06:30,943
I'M NOT GOING TO GO INTO THAT 

5382
04:06:30,943 --> 04:06:31,143
STORY.

5383
04:06:31,143 --> 04:06:32,611
I WANT TO GO INTO A RATHER 

5384
04:06:32,611 --> 04:06:34,480
DIFFERENT STORY.

5385
04:06:34,480 --> 04:06:38,350
THIS JUST SHOWS A VERY, VERY 

5386
04:06:38,350 --> 04:06:44,890
PARTIAL TREE, IF YOU LIKE, 

5387
04:06:44,890 --> 04:06:46,825
PHILOGENY OF ENDOGENOUS 

5388
04:06:46,825 --> 04:06:47,960
RETROVIRUSES COMPARED TO THE 

5389
04:06:47,960 --> 04:06:49,061
RELATED EXOGENOUS.

5390
04:06:49,061 --> 04:06:51,096
YOU CAN SEE THE X IS MARKED 

5391
04:06:51,096 --> 04:06:52,264
EXOGENOUS VIRUSES.

5392
04:06:52,264 --> 04:06:54,333
THE BLUE SQUARES MARK ONES THAT 

5393
04:06:54,333 --> 04:06:56,668
ARE FOUND IN HUMANS, ONLY A VERY

5394
04:06:56,668 --> 04:06:57,903
SMALL FRACTION OF THE 80,000 OR 

5395
04:06:57,903 --> 04:06:59,204
SO THAT WE CARRY.

5396
04:06:59,204 --> 04:07:00,572
AS YOU CAN SEE, THERE JUST HAVE 

5397
04:07:00,572 --> 04:07:02,841
BEEN LOTS AND LOTS OF THESE.

5398
04:07:02,841 --> 04:07:04,610
EACH OF THESE, WE CONSIDER, 

5399
04:07:04,610 --> 04:07:10,482
REPRESENTS A WAVE OF INJECTION 

5400
04:07:10,482 --> 04:07:14,853
THAT A PANZOOTIC INFECTION THAT 

5401
04:07:14,853 --> 04:07:16,955
IT CAME AND THEN WENT EXTINCT 

5402
04:07:16,955 --> 04:07:20,058
AND IS LEFT TO US ONLY BY THIS 

5403
04:07:20,058 --> 04:07:22,828
REMARKABLE FOSSIL RECORD.

5404
04:07:22,828 --> 04:07:25,230
A FAIR AMOUNT OF THE WORK OF MY 

5405
04:07:25,230 --> 04:07:26,798
LABORATORY UP IN BOSTON IS 

5406
04:07:26,798 --> 04:07:28,600
DEVOTED TO TRY TO INTERPRET 

5407
04:07:28,600 --> 04:07:28,834
THIS.

5408
04:07:28,834 --> 04:07:31,170
I'M GOING TO TELL YOU 

5409
04:07:31,170 --> 04:07:32,437
PARTICULARLY ONE PRO VIRUS AT 

5410
04:07:32,437 --> 04:07:34,840
ONE LOCATION, THAT'S IN A GROUP 

5411
04:07:34,840 --> 04:07:42,447
OF VIRUSES CALLED HERV-K 

5412
04:07:42,447 --> 04:07:43,081
(HML-2).

5413
04:07:43,081 --> 04:07:44,516
IT'S THE MOST RECENTLY ACQUIRED 

5414
04:07:44,516 --> 04:07:46,418
AND THE MOST RECENTLY EXTINCT 

5415
04:07:46,418 --> 04:07:46,652
VIRUS.

5416
04:07:46,652 --> 04:07:49,021
THESE VIRUSES WERE STILL BEING 

5417
04:07:49,021 --> 04:07:50,722
INTRODUCED INTO THE HUMAN GENOME

5418
04:07:50,722 --> 04:07:52,257
UP UNTIL MAYBE AS LITTLE AS HALF

5419
04:07:52,257 --> 04:07:53,292
A MILLION YEARS AGO.

5420
04:07:53,292 --> 04:07:54,760
AS FAR AS WE COULD TELL, THERE 

5421
04:07:54,760 --> 04:07:56,795
WERE NO MORE INTRODUCTIONS SINCE

5422
04:07:56,795 --> 04:07:57,095
THEN.

5423
04:07:57,095 --> 04:07:59,598
AS I SAID AGAIN, THEY'VE ALL 

5424
04:07:59,598 --> 04:08:08,907
GONE EXTINCT.

5425
04:08:08,907 --> 04:08:10,809
SO THERE ARE ABOUT 100 PRO 

5426
04:08:10,809 --> 04:08:11,210
VIRUSES.

5427
04:08:11,210 --> 04:08:13,879
EACH OF US CARRIES ABOUT 1 HOW 

5428
04:08:13,879 --> 04:08:15,380
MANY OF THESE PROVIRUSES, MOST 

5429
04:08:15,380 --> 04:08:16,949
OF THEM IDENTICAL FROM ONE OF US

5430
04:08:16,949 --> 04:08:19,184
TO ANOTHER, AT LEAST IN THEIR 

5431
04:08:19,184 --> 04:08:19,451
LOCATION.

5432
04:08:19,451 --> 04:08:21,019
THERE ARE A FEW THAT ARE 

5433
04:08:21,019 --> 04:08:21,820
POLYMORPHIC IN LOCATION.

5434
04:08:21,820 --> 04:08:24,356
THIS IS THE ONLY GROUP OF 

5435
04:08:24,356 --> 04:08:26,124
ENDOGENOUS VIRUSES IN HUMANS, AS

5436
04:08:26,124 --> 04:08:28,360
FAR AS WE KNOW, THAT HAS 

5437
04:08:28,360 --> 04:08:29,895
POLYMORPHIC INSERTION OF THAT 

5438
04:08:29,895 --> 04:08:30,095
SORT.

5439
04:08:30,095 --> 04:08:31,563
I HAVE A FEW THAT YOU MIGHT NOT 

5440
04:08:31,563 --> 04:08:33,031
HAVE, AND EACH OF YOU MAY HAVE A

5441
04:08:33,031 --> 04:08:34,099
FEW THAT I DON'T HAVE.

5442
04:08:34,099 --> 04:08:36,034
AS YOU CAN SEE FROM LOOKING AT 

5443
04:08:36,034 --> 04:08:38,437
THESE, THESE ARE JUST GENOME 

5444
04:08:38,437 --> 04:08:45,177
MAPS REFERRED, MANY OF THEM BY 

5445
04:08:45,177 --> 04:08:47,112
PULLING THEM OFF A HUMAN GENOME 

5446
04:08:47,112 --> 04:08:48,146
SEQUENCE EARLY ON.

5447
04:08:48,146 --> 04:08:51,750
MANY OF THEM ARE DEFECTIVE.

5448
04:08:51,750 --> 04:08:53,485
ALL OF THEM ARE DEFECTIVE IN ONE

5449
04:08:53,485 --> 04:08:56,755
WAY OR ANOTHER, MANY OF THESE BY

5450
04:08:56,755 --> 04:08:59,324
DELETIONS AND STOP CODON 

5451
04:08:59,324 --> 04:09:01,193
INSERTIONS AND SO ON AND SO 

5452
04:09:01,193 --> 04:09:01,460
FORTH.

5453
04:09:01,460 --> 04:09:03,195
I WANT TO FOE UBIQUITOUS YOUR 

5454
04:09:03,195 --> 04:09:04,529
ATTENTION TO TWO OF THESE.

5455
04:09:04,529 --> 04:09:06,498
THE ONE I'M REALLY INTERESTED IN

5456
04:09:06,498 --> 04:09:11,870
IS THE ONE AT CHROMOSOME 3Q12.3.

5457
04:09:11,870 --> 04:09:15,207
WE IDENTIFY THESE BY THEIR 

5458
04:09:15,207 --> 04:09:15,974
CHROMOSOMAL BAND POSITION.

5459
04:09:15,974 --> 04:09:17,809
THERE'S PRETTY MUCH ONLY ONE PER

5460
04:09:17,809 --> 04:09:18,944
BAND, MAYBE TWO OR THREE.

5461
04:09:18,944 --> 04:09:22,447
IN FACT, WE FIND THAT IN MUCH 

5462
04:09:22,447 --> 04:09:23,749
BETTER WAYS THAN ANY OF THE 

5463
04:09:23,749 --> 04:09:25,350
PREVIOUS NOMENCLATURES.

5464
04:09:25,350 --> 04:09:27,686
WE'RE NOT QUITE ALONE IN THIS, A

5465
04:09:27,686 --> 04:09:30,088
LOT OF PEOPLE PICK IT UP.

5466
04:09:30,088 --> 04:09:31,957
BUT WE STILL USE OLD 

5467
04:09:31,957 --> 04:09:32,291
NOMENCLATURES.

5468
04:09:32,291 --> 04:09:33,525
IF YOU READ A PAPER ON THIS 

5469
04:09:33,525 --> 04:09:35,060
SUBJECT, YOU NEED A TRANSLATOR 

5470
04:09:35,060 --> 04:09:35,761
WITH YOU.

5471
04:09:35,761 --> 04:09:37,829
FORTUNATELY, THIS PAPER ACTUALLY

5472
04:09:37,829 --> 04:09:39,031
SERVES AS THIS BECAUSE WE HAVE 

5473
04:09:39,031 --> 04:09:40,465
SYNONYMS NEXT TO ALL OF THESE.

5474
04:09:40,465 --> 04:09:41,800
AND THE OTHER PROVIRUS I'M GOING

5475
04:09:41,800 --> 04:09:46,371
TO MENTION IS THIS ONE AT 

5476
04:09:46,371 --> 04:09:47,706
22Q11.23.

5477
04:09:47,706 --> 04:09:49,041
NOW, YOU'LL NOTE I HAVE THESE IN

5478
04:09:49,041 --> 04:09:50,909
DIFFERENT COLORS, AND THAT'S 

5479
04:09:50,909 --> 04:09:52,611
BECAUSE, IF YOU LOOK AT MY LAST 

5480
04:09:52,611 --> 04:09:55,280
POINT THERE, THIS GROUP OF 

5481
04:09:55,280 --> 04:10:01,653
VIRUSES, HERV-K (HML-2) GROUP 

5482
04:10:01,653 --> 04:10:04,656
CAN BE SUBDIVIDED INTO THREE 

5483
04:10:04,656 --> 04:10:13,965
TAXA, LTRSA, 5B, AND 5HS.

5484
04:10:13,965 --> 04:10:16,868
THE OTHER TWO TAXA, THEY'RE 

5485
04:10:16,868 --> 04:10:18,804
KNOWN INTEGRATIONS FOR MORE THAN

5486
04:10:18,804 --> 04:10:20,339
THE LAST 5 MILLION YEARS HAVE 

5487
04:10:20,339 --> 04:10:21,606
FOUND THEIR WAY INTO THE 

5488
04:10:21,606 --> 04:10:21,873
GERMLINE.

5489
04:10:21,873 --> 04:10:23,442
IF YOU LOOK AT THE THREE ON THE 

5490
04:10:23,442 --> 04:10:25,811
LEFT, THE ONES IN BLUE NOW JUST 

5491
04:10:25,811 --> 04:10:28,613
FOR THE GAG GENE, BUT ANY OTHER 

5492
04:10:28,613 --> 04:10:30,816
TREE WOULD LOOK THE SAME.

5493
04:10:30,816 --> 04:10:36,254
THE ONES IN BLUE ARE THE MOST 

5494
04:10:36,254 --> 04:10:37,723
RECENT, AS YOU CAN TELL IN THE 

5495
04:10:37,723 --> 04:10:39,458
TERMINAL, THE BRANCH ON THESE, 

5496
04:10:39,458 --> 04:10:41,426
BRANCH LENGTHS ON THESE IS MORE 

5497
04:10:41,426 --> 04:10:42,761
OR LESS PROPORTIONAL TO THE AGE 

5498
04:10:42,761 --> 04:10:45,263
OF THE PROVIRUS, AND THESE GO 

5499
04:10:45,263 --> 04:10:48,133
BACK MANY MILLIONS, 25 OR MORE 

5500
04:10:48,133 --> 04:10:51,303
MILLION YEARS.

5501
04:10:51,303 --> 04:10:53,438
THE 5B PROVIRUSES OR THE OTHER 

5502
04:10:53,438 --> 04:10:55,907
I'LL CALL YOUR ATTENTION TO IN 

5503
04:10:55,907 --> 04:10:58,710
PINK, AND THEY ARE -- THEY 

5504
04:10:58,710 --> 04:11:00,345
APPEAR TO BE ANCESTRAL TO THE 

5505
04:11:00,345 --> 04:11:03,348
OTHER TWO GROUPS.  THE 5A I'M 

5506
04:11:03,348 --> 04:11:04,750
NOT GOING TO TALK ABOUT DOWN AT 

5507
04:11:04,750 --> 04:11:09,121
THE BOTTOM, THE GREEN ONES.

5508
04:11:09,121 --> 04:11:13,492
NOW, ALL OF THESE, I SAID, ARE 

5509
04:11:13,492 --> 04:11:13,759
DEFECTIVE.

5510
04:11:13,759 --> 04:11:17,062
THEY ALL HAVE VARIOUS KINDS OF 

5511
04:11:17,062 --> 04:11:17,963
DELETIONS.

5512
04:11:17,963 --> 04:11:22,501
PAUL'S LAB AND ALSO TERRY 

5513
04:11:22,501 --> 04:11:24,369
HINMAN'S LAB IN FRANCE, HAD THE 

5514
04:11:24,369 --> 04:11:26,104
ORIGINAL INSIGHT THE VIRUS THAT 

5515
04:11:26,104 --> 04:11:27,672
GAVE RISE TO THE RECENT ONES IS 

5516
04:11:27,672 --> 04:11:29,574
PRETTY MUCH ESSENTIALLY THE 

5517
04:11:29,574 --> 04:11:31,576
SAME, AND MOST OF THESE 

5518
04:11:31,576 --> 04:11:33,478
MUTATIONS HAVE ARISEN IN THE 

5519
04:11:33,478 --> 04:11:34,980
MANY MILLIONS OF YEARS FOR THE 

5520
04:11:34,980 --> 04:11:37,315
MOST PART SINCE THEY WERE 

5521
04:11:37,315 --> 04:11:38,183
INTEGRATED.

5522
04:11:38,183 --> 04:11:40,952
THEREFORE, THE CONSENSUS 

5523
04:11:40,952 --> 04:11:42,888
SEQUENCE BETWEEN ALL OF THOSE 

5524
04:11:42,888 --> 04:11:43,722
SHOULD REPRESENT SOMETHING CLOSE

5525
04:11:43,722 --> 04:11:44,456
TO THE ORIGINAL VIRUS.

5526
04:11:44,456 --> 04:11:46,892
SO THEY CREATED THIS VIRUS 

5527
04:11:46,892 --> 04:11:53,432
CALLED KCON, WHICH I'LL USE A 

5528
04:11:53,432 --> 04:11:54,833
LOT, WHICH WAS DONE BY TAKING 

5529
04:11:54,833 --> 04:11:57,169
THE CONSENSUS OF ALL OF THE 

5530
04:11:57,169 --> 04:11:59,671
SEQUENCES SHOWN UP AT THE TOP 

5531
04:11:59,671 --> 04:12:02,207
AND PUT THAT ON A NON-ROOTED 

5532
04:12:02,207 --> 04:12:03,675
TREE, YOU CAN SEE THAT SITS MORE

5533
04:12:03,675 --> 04:12:05,610
OR LESS AT THE CENTER, WHICH IS 

5534
04:12:05,610 --> 04:12:16,154
WHERE YOU WOULD EXPECT IT TO BE.

5535
04:12:16,721 --> 04:12:18,123
THAT ACTUALLY CREATES AN 

5536
04:12:18,123 --> 04:12:18,623
INFECTIOUS VIRUS.

5537
04:12:18,623 --> 04:12:21,860
IF YOU JUST REFLECT THE GAG OR 

5538
04:12:21,860 --> 04:12:26,198
GAG GENES, YOU GET THIS NICE 

5539
04:12:26,198 --> 04:12:27,399
VIRUS LIKE PARTICLES.

5540
04:12:27,399 --> 04:12:29,534
SO THAT ACTUALLY HAS BEEN 

5541
04:12:29,534 --> 04:12:31,770
EXTREMELY USEFUL TO US FOR THE 

5542
04:12:31,770 --> 04:12:32,137
EXPERIMENTATION.

5543
04:12:32,137 --> 04:12:36,541
I DON'T KNOW WHAT TO TELL YOU.

5544
04:12:36,541 --> 04:12:39,478
YOU CAN AGE THE INSERTION OF 

5545
04:12:39,478 --> 04:12:41,847
ENDOGENOUS PROVIRUS BY LOOKING 

5546
04:12:41,847 --> 04:12:46,451
AT PHILOGENETIC DISTRIBUTION FOR

5547
04:12:46,451 --> 04:12:48,286
THE ONES IT'S IN, OR ONES THAT 

5548
04:12:48,286 --> 04:12:49,688
ARE OLD ENOUGH TO BE IN THE 

5549
04:12:49,688 --> 04:12:49,921
SPECIES.

5550
04:12:49,921 --> 04:12:52,157
THESE TWO PROVIRUSES, AS YOU CAN

5551
04:12:52,157 --> 04:12:56,194
SEE, ARE ALSO FOUND IN 

5552
04:12:56,194 --> 04:12:59,397
CHIMPANZEES, AND IN PARTICULAR 

5553
04:12:59,397 --> 04:13:04,402
IN GORILLAS, BUT NOT IN 

5554
04:13:04,402 --> 04:13:04,736
ORANGUTANS.

5555
04:13:04,736 --> 04:13:06,571
THAT DATES THE COMMON ANCESTOR 

5556
04:13:06,571 --> 04:13:08,640
AND PROBABLY ALMOST CERTAINLY 

5557
04:13:08,640 --> 04:13:10,675
THE INTEGRATION GAVE RISE TO 

5558
04:13:10,675 --> 04:13:12,277
THIS, BACK INTO SOME COMMON 

5559
04:13:12,277 --> 04:13:15,280
ANCESTOR OF HUMANS AND GORILLAS.

5560
04:13:15,280 --> 04:13:17,649
THAT IS ON THE ORDER OF 8 TO 12 

5561
04:13:17,649 --> 04:13:19,184
MILLION YEARS AGO.

5562
04:13:19,184 --> 04:13:21,620
WE'LL SEE -- IN ROUND NUMBERS, 

5563
04:13:21,620 --> 04:13:23,588
WE'LL SAY 10 MILLION YEARS AGO.

5564
04:13:23,588 --> 04:13:24,389
THESE ARE ABOUT 10 MILLION YEARS

5565
04:13:24,389 --> 04:13:31,997
OLD.

5566
04:13:31,997 --> 04:13:38,169
THE 3Q 12.3 HAS AN INTERESTING 

5567
04:13:38,169 --> 04:13:38,470
PROPERTIES.

5568
04:13:38,470 --> 04:13:39,771
THE STRUCTURE OF THE VIRUS, THE 

5569
04:13:39,771 --> 04:13:41,239
GAG GENE IS COMPLETELY INTACT.

5570
04:13:41,239 --> 04:13:42,707
IT'S BEEN PRESERVED INTACT FOR 

5571
04:13:42,707 --> 04:13:45,443
10 MILLION YEARS SITTING IN THE 

5572
04:13:45,443 --> 04:13:45,944
GENOME.

5573
04:13:45,944 --> 04:13:47,045
HOWEVER, IT WAS DERIVED -- IF 

5574
04:13:47,045 --> 04:13:49,447
YOU LOOK AT THAT, THERE WAS A 

5575
04:13:49,447 --> 04:13:50,815
RECOMBINATION EVENT THAT 

5576
04:13:50,815 --> 04:13:51,883
GENERATED THIS PROVIRUS 

5577
04:13:51,883 --> 04:13:54,185
ORIGINALLY THAT BROUGHT IN THIS 

5578
04:13:54,185 --> 04:13:59,057
OLDER TYPE B PROVIRUS, AND 

5579
04:13:59,057 --> 04:14:00,492
CROSSOVER WAS IN THE MIDDLE OF 

5580
04:14:00,492 --> 04:14:06,364
THE GAG TO THE MIDDLE OF POL.

5581
04:14:06,364 --> 04:14:08,400
THAT WAS ALMOST IDENTICAL 

5582
04:14:08,400 --> 04:14:09,834
BETWEEN THE TWO PROVIRUSES.

5583
04:14:09,834 --> 04:14:17,142
THE OTHER PROVIRUS, THE 22Q11.23

5584
04:14:17,142 --> 04:14:19,344
IS STILL IN OUR GENOME AND HAS A

5585
04:14:19,344 --> 04:14:22,080
COMPLETELY INTACT GAG GENE.

5586
04:14:22,080 --> 04:14:23,582
SO THERE'S RATHER STRONG 

5587
04:14:23,582 --> 04:14:24,616
EVIDENCE THERE'S SOMETHING 

5588
04:14:24,616 --> 04:14:25,584
SELECTIVELY ADVANTAGEOUS ABOUT 

5589
04:14:25,584 --> 04:14:28,286
HAVING THESE GAG GENES IN OUR 

5590
04:14:28,286 --> 04:14:30,388
GENOME.

5591
04:14:30,388 --> 04:14:31,656
ANOTHER INTERESTING PROPERTY IS 

5592
04:14:31,656 --> 04:14:36,661
OF 3Q12.3 IS THAT THE 5 PRIME 

5593
04:14:36,661 --> 04:14:40,699
LTR BUT NOT THE 3 PRIME LTR 

5594
04:14:40,699 --> 04:14:44,102
CONTAINS THE FOUR PAIR BASE 

5595
04:14:44,102 --> 04:14:46,271
MUTATION THAT, AS IT HAPPENS 

5596
04:14:46,271 --> 04:14:50,609
CREATES THE BINDING SITES FOR 

5597
04:14:50,609 --> 04:14:53,278
HOX-PBX TRANSCRIPTION FAMILY 

5598
04:14:53,278 --> 04:14:53,612
PROTEIN.

5599
04:14:53,612 --> 04:14:54,813
THIS IS A COMMON TRANSCRIPTION 

5600
04:14:54,813 --> 04:14:56,481
FACTOR, AND IT APPEARS A LOT OF 

5601
04:14:56,481 --> 04:14:56,681
TIMES.

5602
04:14:56,681 --> 04:14:58,516
IT APPEARS IN CANCER AS 

5603
04:14:58,516 --> 04:14:58,783
IMPORTANT.

5604
04:14:58,783 --> 04:14:59,951
IT APPEARS IN LOTS OF 

5605
04:14:59,951 --> 04:15:08,026
DIFFERENTIATION AS IMPORTANT.

5606
04:15:08,026 --> 04:15:11,329
AS I WILL SHOW YOU IN THIS 

5607
04:15:11,329 --> 04:15:13,698
SLIDE, THIS IS DONE SOME YEARS 

5608
04:15:13,698 --> 04:15:17,002
AGO BY MEGAN IN MY LAB, IF YOU 

5609
04:15:17,002 --> 04:15:18,970
LOOK AT -- WHAT SHE DID IS JUST 

5610
04:15:18,970 --> 04:15:21,006
COMPARED WHAT THAT TRANSCRIPTION

5611
04:15:21,006 --> 04:15:23,308
FACTOR DOES IN A STANDARD 

5612
04:15:23,308 --> 04:15:24,809
PROMOTER BASHING ASSAY AND PUTS 

5613
04:15:24,809 --> 04:15:28,880
IT NEXT TO A VOCIFEROUS GENE.

5614
04:15:28,880 --> 04:15:37,055
IF YOU LOOK AT THE BOTTOM, THE

5615
04:15:37,055 --> 04:15:41,693
3Q PRIME WILD TYPE LTR, IF YOU 

5616
04:15:41,693 --> 04:15:43,862
MAKE NO OTHER CHANGE, IT BRINGS 

5617
04:15:43,862 --> 04:15:45,664
DOWN THE EXPRESSION A LOT.

5618
04:15:45,664 --> 04:15:49,634
ANOTHER RELATED PROPERTY OF THIS

5619
04:15:49,634 --> 04:15:54,005
PROVIRUS WAS WORKED OUT BY AIDAN

5620
04:15:54,005 --> 04:15:55,440
BURN A FEW YEARS AGO WHEN HE 

5621
04:15:55,440 --> 04:15:57,575
LOOKED AT THE EXPRESSION OF ALL 

5622
04:15:57,575 --> 04:16:00,111
ENDOGENOUS PROVIRUSES OF THESE 

5623
04:16:00,111 --> 04:16:08,687
GROUPS ACROSS THE LARGE GTEX 

5624
04:16:08,687 --> 04:16:12,223
DATABASE OF LARGE RNASEQ DATA 

5625
04:16:12,223 --> 04:16:15,360
FROM TISSUES FROM 100 OR MORE 

5626
04:16:15,360 --> 04:16:20,699
INDIVIDUALS, AND AS YOU CAN SEE,

5627
04:16:20,699 --> 04:16:22,167
3Q12.3 IS ONE THAT'S BY FAR THE 

5628
04:16:22,167 --> 04:16:23,601
MOST HIGHLY EXPRESSED.

5629
04:16:23,601 --> 04:16:26,604
THE ONE EXPRESSED MORE IS HIGHLY

5630
04:16:26,604 --> 04:16:26,871
DEFECTIVE.

5631
04:16:26,871 --> 04:16:28,106
IT DOES SOMETHING, BUT IT HAS 

5632
04:16:28,106 --> 04:16:30,608
NOTHING TO DO WITH BEING A 

5633
04:16:30,608 --> 04:16:31,342
VIRUS.

5634
04:16:31,342 --> 04:16:37,849
THIS HAS A TECH ADD GENE, AND 

5635
04:16:37,849 --> 04:16:39,551
THAT GENE IS HIGHLY SPECIFIC, 

5636
04:16:39,551 --> 04:16:41,286
NOT IN CHIMPANZEES.

5637
04:16:41,286 --> 04:16:43,088
SO IT OCCURRED IN THE LAST 5 

5638
04:16:43,088 --> 04:16:43,855
MILLION YEARS.

5639
04:16:43,855 --> 04:16:46,458
AND THAT MUTATION FURTHER, THAT 

5640
04:16:46,458 --> 04:16:49,427
MUTATION FURTHER IS FIXED IN THE

5641
04:16:49,427 --> 04:16:58,937
HUMAN GENOME.

5642
04:16:58,937 --> 04:17:00,271
THE OTHER THING THAT IS 

5643
04:17:00,271 --> 04:17:01,639
REMARKABLE IS THAT GAG -- IF YOU

5644
04:17:01,639 --> 04:17:05,009
LOOK AT THAT GAG GENE ACROSS 

5645
04:17:05,009 --> 04:17:11,349
5,000 ALLELES IN THE HUMAN 1,000

5646
04:17:11,349 --> 04:17:14,085
GENOME DATABASE, THE CAPSID 

5647
04:17:14,085 --> 04:17:17,422
DOMAIN HAS NOT A SINGLE 

5648
04:17:17,422 --> 04:17:17,689
MUTATION.

5649
04:17:17,689 --> 04:17:22,494
THERE'S NO SINGLE CHANGE IN THE 

5650
04:17:22,494 --> 04:17:25,430
CAPSID DOMAIN.

5651
04:17:25,430 --> 04:17:26,731
THERE ARE VERY FEW OVERALL, AND 

5652
04:17:26,731 --> 04:17:27,966
IT'S OPEN TO EVERYBODY.

5653
04:17:27,966 --> 04:17:29,601
THAT FOR US IS RATHER STRONG 

5654
04:17:29,601 --> 04:17:31,569
EVIDENCE THAT THIS IS DOING 

5655
04:17:31,569 --> 04:17:32,203
SOMETHING INTERESTING AND MAYBE 

5656
04:17:32,203 --> 04:17:35,306
EVEN IMPORTANT.

5657
04:17:35,306 --> 04:17:36,441
SO OUR HYPOTHESIS GIVEN WHAT 

5658
04:17:36,441 --> 04:17:38,209
I'VE TOLD YOU BEFORE ABOUT 

5659
04:17:38,209 --> 04:17:40,245
ENDOGENOUS PROVIRUSES IS THAT 

5660
04:17:40,245 --> 04:17:41,946
THIS MAY HAVE EVOLVED INTO SOME 

5661
04:17:41,946 --> 04:17:44,415
KIND OF RESTRICTION FACTOR FOR 

5662
04:17:44,415 --> 04:17:46,818
REPLICATION OF THAT RELATED 

5663
04:17:46,818 --> 04:17:50,355
VIRUS, AND OUR SPECULATIVE 

5664
04:17:50,355 --> 04:17:51,189
HYPOTHESIS, BECAUSE WE'LL NEVER 

5665
04:17:51,189 --> 04:17:53,658
BE ABLE TO ACTUALLY PROVE IS, IS

5666
04:17:53,658 --> 04:17:54,726
THAT WE HAVE EVIDENCE OF THIS 

5667
04:17:54,726 --> 04:17:57,595
GROUP OF PROVIRUS, THIS GROUP OF

5668
04:17:57,595 --> 04:17:59,664
VIRUSES WENT EXTINCT AS A VIRUS 

5669
04:17:59,664 --> 04:18:02,467
ON THE ORDER OF HALF A MILLION 

5670
04:18:02,467 --> 04:18:04,669
YEARS AGO.

5671
04:18:04,669 --> 04:18:06,137
OUR SPECULATIVE HYPOTHESIS IS IT

5672
04:18:06,137 --> 04:18:08,573
WAS THE ORIGIN OF THIS MUTATION 

5673
04:18:08,573 --> 04:18:09,574
AND ITS SELECTION THAT SPREAD 

5674
04:18:09,574 --> 04:18:11,009
THROUGH THE POPULATION THAT 

5675
04:18:11,009 --> 04:18:12,277
ACTUALLY LED TO THE EXTINCTION 

5676
04:18:12,277 --> 04:18:12,811
OF THE VIRUS.

5677
04:18:12,811 --> 04:18:14,112
I'LL JUST LEAVE YOU WITH THAT AS

5678
04:18:14,112 --> 04:18:20,351
A SPECULATION RIGHT NOW.

5679
04:18:20,351 --> 04:18:22,687
SO THE REST OF WHAT I'LL TELL 

5680
04:18:22,687 --> 04:18:24,923
YOU IS ABOUT TRYING TO PROVE 

5681
04:18:24,923 --> 04:18:27,425
THIS OR TEST THIS HYPOTHESIS AND

5682
04:18:27,425 --> 04:18:29,194
THEN CHECK IF IT TESTS OUT 

5683
04:18:29,194 --> 04:18:30,762
BECAUSE YOU MIGHT GUESS IT DID, 

5684
04:18:30,762 --> 04:18:39,137
CHECK THE MECHANISM OF IT.

5685
04:18:39,137 --> 04:18:41,573
THE GENERAL EXPERIMENT IS TO 

5686
04:18:41,573 --> 04:18:46,344
EXPRESS THE KCON, EITHER 

5687
04:18:46,344 --> 04:18:50,415
PSEUDOVIRUS OR EXPRESSIVE VIRUS 

5688
04:18:50,415 --> 04:18:53,418
BY ADDING AN EXTRA GENE AND TEST

5689
04:18:53,418 --> 04:18:57,121
WITH THE 3Q12.3 GAG GENE BECAUSE

5690
04:18:57,121 --> 04:18:58,256
THE GAG GENE IS MOST CERTAINLY 

5691
04:18:58,256 --> 04:19:00,158
DOING THE ACTION HERE, IF IT'S 

5692
04:19:00,158 --> 04:19:05,129
GOING TO DO ANYTHING, AND 

5693
04:19:05,129 --> 04:19:07,065
CO-TRANSEFFECT AND CHECK FOR 

5694
04:19:07,065 --> 04:19:08,466
NON-INFECTIOUS VIRUSES, OR IF 

5695
04:19:08,466 --> 04:19:10,068
YOU ADD AN ENVELOPE GENE, TO 

5696
04:19:10,068 --> 04:19:11,069
ACTUALLY LOOK FOR AN INFECTIOUS 

5697
04:19:11,069 --> 04:19:13,371
VIRUS.

5698
04:19:13,371 --> 04:19:14,505
AT THE SAME TIME, WE CAN DO LOTS

5699
04:19:14,505 --> 04:19:17,275
OF OTHER STUDIES TO SEE WHAT'S 

5700
04:19:17,275 --> 04:19:17,542
GOING ON.

5701
04:19:17,542 --> 04:19:20,144
THIS CURVE IS PLOTTED PERHAPS A 

5702
04:19:20,144 --> 04:19:22,614
LITTLE BIT BACKWARDS, BUT YOU'LL

5703
04:19:22,614 --> 04:19:27,352
NOTICE NOW AT THE -- SO THIS 

5704
04:19:27,352 --> 04:19:36,394
SHOWS THAT THE HERV-K, THE 

5705
04:19:36,394 --> 04:19:37,362
3Q12.3 RATHER STRONGLY BLOCKS 

5706
04:19:37,362 --> 04:19:38,796
PRODUCTION, IN THIS CASE FOR AN 

5707
04:19:38,796 --> 04:19:41,299
INFECTIOUS VIRUS, AS YOU'LL SEE 

5708
04:19:41,299 --> 04:19:43,034
THE SAME IF YOU LOOK FOR 

5709
04:19:43,034 --> 04:19:45,236
PARTICLE FORMATION, AND UP TO 

5710
04:19:45,236 --> 04:19:48,506
ABOUT -- I'M GOING BACKWARDS 

5711
04:19:48,506 --> 04:19:50,675
THERE.

5712
04:19:50,675 --> 04:19:58,182
UP TO ABOUT 1 IN 16 IS ENOUGH TO

5713
04:19:58,182 --> 04:20:00,418
GIVE ABOUT A 50 PERCENT 

5714
04:20:00,418 --> 04:20:00,718
INHIBITION.

5715
04:20:00,718 --> 04:20:05,156
ONE GAG MOLECULE PER 16 OR 

5716
04:20:05,156 --> 04:20:15,700
ONE -- OR ABOUT PER 3 HEXAMERS, 

5717
04:20:18,636 --> 04:20:21,506
IF YOU LIKE, IS ABOUT THE RATIO,

5718
04:20:21,506 --> 04:20:22,941
AND SOME VIRUSES YOU GET EVEN 

5719
04:20:22,941 --> 04:20:24,842
MORE.

5720
04:20:24,842 --> 04:20:27,078
JUST TO REMIND EVERYBODY OF 

5721
04:20:27,078 --> 04:20:29,247
ERIC'S VERY POPULAR SLIDE, ON 

5722
04:20:29,247 --> 04:20:31,049
THE TINY CORNER OF THIS SLIDE, 

5723
04:20:31,049 --> 04:20:32,650
JUST TO MAKE SURE WE ALL 

5724
04:20:32,650 --> 04:20:33,618
UNDERSTAND -- I'M SURE YOU ALL 

5725
04:20:33,618 --> 04:20:35,119
KNOW THIS ACTUALLY.

5726
04:20:35,119 --> 04:20:38,356
BUT THE GAG CONSISTS OF TOLEDO 

5727
04:20:38,356 --> 04:20:42,060
MAINS, MATRIX, CAPSID, AND 

5728
04:20:42,060 --> 04:20:46,431
NUCLEOCAPSID PRINCIPALLY, PLUS 

5729
04:20:46,431 --> 04:20:48,366
SOME OTHERS INVOLVED WITH MATRIX

5730
04:20:48,366 --> 04:20:49,867
DOMAINS, BUT I'M NOT GOING TO 

5731
04:20:49,867 --> 04:20:50,868
TALK ABOUT THAT.

5732
04:20:50,868 --> 04:20:53,871
THAT GOES WITH MATRIX ASSEMBLING

5733
04:20:53,871 --> 04:20:58,977
THE MEMBRANE, AND CAPSIDS 

5734
04:20:58,977 --> 04:21:01,713
ASSEMBLING EACH OTHER INTO 

5735
04:21:01,713 --> 04:21:04,082
HEXAMERS GENERALLY, AND THEN 

5736
04:21:04,082 --> 04:21:05,917
WRAPPING THE MEMBRANE AROUND, 

5737
04:21:05,917 --> 04:21:08,720
BUDDING OCCURS, AND IT BUDS OFF 

5738
04:21:08,720 --> 04:21:10,922
AFTER INCLUDING MACHINERY TO 

5739
04:21:10,922 --> 04:21:13,091
CLOSE OFF THE MEMBRANE AT THE 

5740
04:21:13,091 --> 04:21:13,391
END.

5741
04:21:13,391 --> 04:21:16,661
THE BUDS IN THE MATURE PARTICLE 

5742
04:21:16,661 --> 04:21:17,895
AND ONLY AFTERWARDS GETS 

5743
04:21:17,895 --> 04:21:18,563
PROCESSED INTO ITS FINAL MATURE 

5744
04:21:18,563 --> 04:21:20,331
STRUCTURE.

5745
04:21:20,331 --> 04:21:22,367
WHAT IS QUITE REMARKABLE IS THAT

5746
04:21:22,367 --> 04:21:23,001
THE DIFFERENCE IN STRUCTURE 

5747
04:21:23,001 --> 04:21:24,836
BETWEEN THE MATURE AND THE 

5748
04:21:24,836 --> 04:21:26,671
IMMATURE, THAT WAS ALREADY 

5749
04:21:26,671 --> 04:21:28,573
MENTIONED A LITTLE BIT THAT T

5750
04:21:28,573 --> 04:21:32,510
THE -- THAT THERE'S QUITE A 

5751
04:21:32,510 --> 04:21:34,846
SUBSTANTIAL REARRANGEMENT 

5752
04:21:34,846 --> 04:21:38,549
BETWEEN HEXAMERS, BUT THEY'RE 

5753
04:21:38,549 --> 04:21:40,585
QUITE A MATURE ORGANIZATION.

5754
04:21:40,585 --> 04:21:42,720
IT'S STRIKING THAT THE IMMATURE 

5755
04:21:42,720 --> 04:21:43,988
PARTICLES, ALTHOUGH THEY CAN'T 

5756
04:21:43,988 --> 04:21:45,490
BE COMPLETE PARTICLES, FOR 

5757
04:21:45,490 --> 04:21:47,859
REASONS I'M NOT GOING TO GO 

5758
04:21:47,859 --> 04:21:49,961
INTO, BUT THEY'RE ALWAYS ALMOST 

5759
04:21:49,961 --> 04:21:51,662
PERFECTLY SPHERICAL IN THE 

5760
04:21:51,662 --> 04:21:52,830
APPEARANCE, IN THE MICROGRAPH, 

5761
04:21:52,830 --> 04:21:54,065
AND YOU'VE ALREADY SEEN SOME OF 

5762
04:21:54,065 --> 04:21:54,665
THAT.

5763
04:21:54,665 --> 04:21:56,667
WHEREAS THE MATURE PARTICLES 

5764
04:21:56,667 --> 04:21:59,037
HAVE THIS -- OFTEN HAVE SOME -- 

5765
04:21:59,037 --> 04:22:06,377
ARE NEVER ACTUALLY PERFECTLY HEX

5766
04:22:06,377 --> 04:22:06,778
GON

5767
04:22:06,778 --> 04:22:07,678
HEXAGONAL SORT OF STRUCTURES 

5768
04:22:07,678 --> 04:22:11,215
THAT YOU SEE WITH MANY OTHER 

5769
04:22:11,215 --> 04:22:11,449
VIRUSES.

5770
04:22:11,449 --> 04:22:13,885
THEY'RE ALWAYS EITHER THIS 

5771
04:22:13,885 --> 04:22:14,519
BULLET-SHAPED CAPSID OR 

5772
04:22:14,519 --> 04:22:16,220
CONE-SHAPED CAPSID THAT YOU SEE 

5773
04:22:16,220 --> 04:22:18,222
IN HIV, OR SOME OTHER STRUCTURE 

5774
04:22:18,222 --> 04:22:20,725
WHICH IS NOT -- WE DON'T -- IT'S

5775
04:22:20,725 --> 04:22:26,164
NOT VERY WELL DEFINED FOR OTHER 

5776
04:22:26,164 --> 04:22:26,497
RETROVIRUSES.

5777
04:22:26,497 --> 04:22:29,300
IT'S KIND OF LUMPY AND UGLY BY 

5778
04:22:29,300 --> 04:22:31,102
AND LARGE.

5779
04:22:31,102 --> 04:22:37,041
IT'S NEVER PERFECT LY ICOSOHEDRL

5780
04:22:37,041 --> 04:22:38,876
IN APPEARANCE.

5781
04:22:38,876 --> 04:22:40,711
THAT'S MADE POSSIBLE BY THE 

5782
04:22:40,711 --> 04:22:42,547
DIFFERENCE BETWEEN THE NTD THAT 

5783
04:22:42,547 --> 04:22:44,582
YOU CAN SEE DOWN IN THE LOWER 

5784
04:22:44,582 --> 04:22:46,851
LEFT THERE BEING -- I'M SORRY, 

5785
04:22:46,851 --> 04:22:48,719
BY THE CAPSID PROTEIN BEING 

5786
04:22:48,719 --> 04:22:50,254
DIVIDED INTO THE N-TERMINAL 

5787
04:22:50,254 --> 04:22:52,290
DOMAIN AND THE C-TERMINAL 

5788
04:22:52,290 --> 04:22:52,723
DOMAIN.

5789
04:22:52,723 --> 04:22:55,426
THE C-TERMINAL DOMAIN OF THE 

5790
04:22:55,426 --> 04:22:58,996
CAPSID INTERACTS CAPSID 

5791
04:22:58,996 --> 04:23:00,264
MOLECULES WITH ONE ANOTHER TO 

5792
04:23:00,264 --> 04:23:04,102
FORM HEXAMERS, AND THE INTERNAL 

5793
04:23:04,102 --> 04:23:08,039
DOMAIN INTERACTS HEXAMER TO 

5794
04:23:08,039 --> 04:23:09,507
HEXAMER TO FORM THE STRUCTURE, 

5795
04:23:09,507 --> 04:23:10,908
AND THAT'S AN OVERSIMPLIFIED 

5796
04:23:10,908 --> 04:23:13,044
VERSION OF WHAT'S GOING ON.

5797
04:23:13,044 --> 04:23:13,911
AND THAT'S MADE POSSIBILITY BY 

5798
04:23:13,911 --> 04:23:15,947
THE STRUCTURE OF THIS LINKER 

5799
04:23:15,947 --> 04:23:17,582
DOMAIN SO THERE CAN BE DIFFERENT

5800
04:23:17,582 --> 04:23:19,650
LEVELS OF INTERACTION BETWEEN 

5801
04:23:19,650 --> 04:23:21,619
DIFFERENT HEXAMERS, AND THAT 

5802
04:23:21,619 --> 04:23:24,655
ALLOWS THE PARTICLES TO ASSUME 

5803
04:23:24,655 --> 04:23:30,561
OTHER THAN THIS ICOSALHEDRAL 

5804
04:23:30,561 --> 04:23:30,828
STRUCTURE.

5805
04:23:30,828 --> 04:23:34,499
AND YOU SEE IT STARTS WITH THE 

5806
04:23:34,499 --> 04:23:35,433
EXPERIMENT IN BLUE THAT STARTS 

5807
04:23:35,433 --> 04:23:39,604
WITH KCON, THE WILD SIDE GAG 

5808
04:23:39,604 --> 04:23:42,874
PROTEIN, AND WITH SUBSTITUTIONS 

5809
04:23:42,874 --> 04:23:45,343
OF THE VARIOUS DOMAINS, CAPSID, 

5810
04:23:45,343 --> 04:23:46,477
NUCLEOCAPSID, AND SO ON.

5811
04:23:46,477 --> 04:23:48,713
IF YOU SUBSTITUTE JUST THE 

5812
04:23:48,713 --> 04:23:51,415
INTERIM PLUS THE LINKER, THEN 

5813
04:23:51,415 --> 04:23:53,918
YOU GET A STRONG INHIBITION, AND

5814
04:23:53,918 --> 04:23:56,020
THE OTHER PARTS YOU SEE LESS 

5815
04:23:56,020 --> 04:24:03,327
INHIBITION OR NO INHIBITION.

5816
04:24:03,327 --> 04:24:05,062
THAT LOCALIZES THE EFFECT OF 

5817
04:24:05,062 --> 04:24:06,330
THIS PARTICULAR REGION.

5818
04:24:06,330 --> 04:24:07,498
WHEN YOU LOOK HARD, PARTICULARLY

5819
04:24:07,498 --> 04:24:09,867
AT THE LINKER SEQUENCE, ONE 

5820
04:24:09,867 --> 04:24:12,136
MUTATION STANDS OUT, AND THAT'S 

5821
04:24:12,136 --> 04:24:20,778
THE DELETION OF THE LUCYRINE.

5822
04:24:20,778 --> 04:24:23,014
AND THAT DELETION IN THE 

5823
04:24:23,014 --> 04:24:26,717
STRUCTURE OF THE IMMATURE KCON 

5824
04:24:26,717 --> 04:24:28,719
PROTEIN FORMS A VERY SPECIFIC 

5825
04:24:28,719 --> 04:24:32,723
HYDROGEN BOND WITHIN A HELICAL 

5826
04:24:32,723 --> 04:24:34,792
REGION IN THE INTERNAL DOMAIN.

5827
04:24:34,792 --> 04:24:36,661
THAT HYDROGEN BOND IS NOT SEEN 

5828
04:24:36,661 --> 04:24:37,762
IN THE MORE MATURE PROTEIN.

5829
04:24:37,762 --> 04:24:38,963
YOU CAN SEE A COUPLE OF OTHERS, 

5830
04:24:38,963 --> 04:24:47,305
BUT NOT THAT ONE.

5831
04:24:47,305 --> 04:24:56,447
IF YOU REPAIR THE SERINE 

5832
04:24:56,447 --> 04:24:58,149
MUTATION -- SORRY.

5833
04:24:58,149 --> 04:25:01,319
IF YOU TAKE THE WILD TYPE VIRUS 

5834
04:25:01,319 --> 04:25:02,553
AND INTRODUCE THESE VARIOUS 

5835
04:25:02,553 --> 04:25:05,156
MUTATIONS INTO IT, YOU FIND ON 

5836
04:25:05,156 --> 04:25:09,427
THE FAR RIGHT INTRODUCING THIS 

5837
04:25:09,427 --> 04:25:11,562
SERINE DELETION RECAPITULATES 

5838
04:25:11,562 --> 04:25:13,231
MOST OF THE ACTIVITY IN THIS 

5839
04:25:13,231 --> 04:25:13,464
PROTEIN.

5840
04:25:13,464 --> 04:25:15,032
NOT QUITE ALL, SO WE THINK THERE

5841
04:25:15,032 --> 04:25:16,667
ARE OTHER MUTATIONS THAT ARE 

5842
04:25:16,667 --> 04:25:18,135
ALSO IMPORTANT, BUT WE HAVEN'T 

5843
04:25:18,135 --> 04:25:19,103
TRACKED THOSE DOWN YET.

5844
04:25:19,103 --> 04:25:20,404
THOSE ARE THE OTHER DIFFERENCES 

5845
04:25:20,404 --> 04:25:23,107
THAT WE SEE REALLY MATTER VERY 

5846
04:25:23,107 --> 04:25:27,211
MUCH AT ALL AS FAR AS THE 

5847
04:25:27,211 --> 04:25:29,480
INHIBITION OF ACTIVITY IS 

5848
04:25:29,480 --> 04:25:29,780
CONCERNED.

5849
04:25:29,780 --> 04:25:31,482
SO WE WANTED TO SEE WHAT THIS 

5850
04:25:31,482 --> 04:25:32,750
LOOKED LIKE IN TERMS OF WHAT THE

5851
04:25:32,750 --> 04:25:34,452
VIRUS IS ACTUALLY DOING AS THE 

5852
04:25:34,452 --> 04:25:36,387
BUDDING IS TRYING TO OCCUR, AS 

5853
04:25:36,387 --> 04:25:39,357
THE REPLICATION IS TRYING TO 

5854
04:25:39,357 --> 04:25:42,159
OCCUR, AND OF COURSE AS THE TAG 

5855
04:25:42,159 --> 04:25:45,062
LINE OF AN OLD JINGLE THAT I 

5856
04:25:45,062 --> 04:25:47,698
THINK YOU HAD TO LIVE IN NEW 

5857
04:25:47,698 --> 04:25:53,304
ENGLAND TO HEAR, IT'S WHO YOU 

5858
04:25:53,304 --> 04:25:53,871
GONNA CALL?

5859
04:25:53,871 --> 04:25:55,106
OF COURSE THE ANSWER IS OBVIOUS,

5860
04:25:55,106 --> 04:25:55,306
RIGHT?

5861
04:25:55,306 --> 04:25:59,677
IT'S ALAN.

5862
04:25:59,677 --> 04:26:02,647
WE CALL OUR VIRUS ASSEMBLY GURU 

5863
04:26:02,647 --> 04:26:08,085
HE

5864
04:26:08,085 --> 04:26:13,624
HERE, AND ALAN CALLS FERRI 

5865
04:26:13,624 --> 04:26:15,493
SOHELLAN, OUR MICROSCOPE EXPERT,

5866
04:26:15,493 --> 04:26:18,763
AND WE SENT ALL THE SAMPLES OF 

5867
04:26:18,763 --> 04:26:19,130
THIS.

5868
04:26:19,130 --> 04:26:22,433
THE FIRST IS ALL THE WILD TYPE 

5869
04:26:22,433 --> 04:26:24,969
SAMPLES, THE KCON WILD TYPE 

5870
04:26:24,969 --> 04:26:27,371
PROTEIN, AND YOU SEE THE 

5871
04:26:27,371 --> 04:26:29,540
PERFECTLY GOOD PARTICLE 

5872
04:26:29,540 --> 04:26:29,807
ASSEMBLY.

5873
04:26:29,807 --> 04:26:32,143
THEY ALWAYS APPEAR PERFECTLY 

5874
04:26:32,143 --> 04:26:36,614
CIRCULAR, IF YOU LIKE PERFECTLY 

5875
04:26:36,614 --> 04:26:39,150
SPHERICAL, AND QUITE UNIFORM IN 

5876
04:26:39,150 --> 04:26:39,917
SIZE.

5877
04:26:39,917 --> 04:26:46,023
UNLIKE THE FINAL PRODUCTS.

5878
04:26:46,023 --> 04:26:49,460
HOWEVER, WHEN YOU LOOK AT 3Q12.3

5879
04:26:49,460 --> 04:26:51,429
GAG ALONE, REMEMBER IS THE 

5880
04:26:51,429 --> 04:26:52,963
MUTANT PROVIRUS, YOU DON'T SEE 

5881
04:26:52,963 --> 04:26:53,931
ANYTHING LIKE THAT.

5882
04:26:53,931 --> 04:26:56,634
YOU JUST SEE THESE SORT OF -- 

5883
04:26:56,634 --> 04:26:58,703
LOOKS LIKE THE THICKENINGS OF 

5884
04:26:58,703 --> 04:27:00,371
THE MEMBRANES WHERE IT'S TRYING 

5885
04:27:00,371 --> 04:27:01,005
TO BUD.

5886
04:27:01,005 --> 04:27:02,373
THEY HAVEN'T ACTUALLY PROVEN 

5887
04:27:02,373 --> 04:27:03,641
YET, WE HAVEN'T DONE ANY 

5888
04:27:03,641 --> 04:27:05,509
EXPERIMENTS TO SHOW DIRECTLY 

5889
04:27:05,509 --> 04:27:07,478
THOSE ARE GAG PROTEINS, BUT IT'S

5890
04:27:07,478 --> 04:27:08,612
HARD TO IMAGINE THEY'RE ANYTHING

5891
04:27:08,612 --> 04:27:11,015
ELSE BECAUSE YOU DON'T SEE THIS 

5892
04:27:11,015 --> 04:27:13,284
EVER UNLESS YOU HAVE THIS IN 

5893
04:27:13,284 --> 04:27:15,019
THERE.

5894
04:27:15,019 --> 04:27:21,826
BUT NOW IF YOU TRANSFECT WITH 

5895
04:27:21,826 --> 04:27:24,995
THE WILD TYPE PROTEINS, PROTEINS

5896
04:27:24,995 --> 04:27:27,631
FROM KCON, WHAT YOU GET ARE WHAT

5897
04:27:27,631 --> 04:27:31,168
WE LIKE TO CALL VUPs, OR VIRUS 

5898
04:27:31,168 --> 04:27:32,970
UNLIKE PARTICLES, AND YOU GET 

5899
04:27:32,970 --> 04:27:35,473
THIS BIZARRE KIND OF ATTEMPT TO 

5900
04:27:35,473 --> 04:27:39,477
BOND BUT NOTHING -- THEY NEVER 

5901
04:27:39,477 --> 04:27:39,677
CLOSE.

5902
04:27:39,677 --> 04:27:41,145
THEY JUST SORT OF GROW AROUND 

5903
04:27:41,145 --> 04:27:51,622
INTO THESE UGLY STRUCTURES.

5904
04:27:56,794 --> 04:27:58,329
IT'S AS THOUGH YOU HAD A MIXTURE

5905
04:27:58,329 --> 04:28:00,598
OF THINGS WITH THOSE THAT CAN'T.

5906
04:28:00,598 --> 04:28:02,032
THAT'S WHAT YOU GET.

5907
04:28:02,032 --> 04:28:03,267
WE HAVEN'T PROVEN THAT'S WHAT IT

5908
04:28:03,267 --> 04:28:03,968
LOOKS LIKE.

5909
04:28:03,968 --> 04:28:05,703
WE HAVE COLLABORATION SET UP TO 

5910
04:28:05,703 --> 04:28:07,037
DO REAL STRUCTURAL ANALYSIS ON 

5911
04:28:07,037 --> 04:28:17,548
THESE THINGS, BUT THAT IS OUR 

5912
04:28:17,748 --> 04:28:18,783
HYPOTHESIS AT LEAST, AND I THINK

5913
04:28:18,783 --> 04:28:20,317
IT'S A PRETTY GOOD ONE, TO 

5914
04:28:20,317 --> 04:28:21,285
UNDERSTAND WHAT'S GOING ON.

5915
04:28:21,285 --> 04:28:22,853
THAT ALLOWS THE FORMATION OF 

5916
04:28:22,853 --> 04:28:24,221
THESE IMMATURE PARTICLES.

5917
04:28:24,221 --> 04:28:31,929
IF YOU DON'T HAVE IT, THEN YOU 

5918
04:28:31,929 --> 04:28:33,864
JUST CAN'T -- THE VIRUS PROBABLY

5919
04:28:33,864 --> 04:28:35,699
CAN'T PULL TOGETHER STRONG 

5920
04:28:35,699 --> 04:28:39,603
ENOUGH TO FOLD A MEMBRANE AROUND

5921
04:28:39,603 --> 04:28:40,671
IT WITHOUT THAT PARTICULAR 

5922
04:28:40,671 --> 04:28:40,905
LINKAGE.

5923
04:28:40,905 --> 04:28:42,840
SO IT STARTS OUT, BUT IT CAN'T 

5924
04:28:42,840 --> 04:28:43,607
GET VERY FAR.

5925
04:28:43,607 --> 04:28:45,443
IF YOU MIX IT WITH GOOD, THEN 

5926
04:28:45,443 --> 04:28:47,211
YOU JUST GET THESE MISSES.

5927
04:28:47,211 --> 04:28:50,014
ERIC ACTUALLY YEARS AGO 

5928
04:28:50,014 --> 04:28:51,382
PUBLISHED WITH HIV WHAT 

5929
04:28:51,382 --> 04:28:59,423
HAPPENS -- THERE'S A HOMOLOGOUS 

5930
04:28:59,423 --> 04:29:03,060
SERINE IS HIV, AND IF YOU LOOK 

5931
04:29:03,060 --> 04:29:05,329
AT THIS, YOU GET THE SIMILAR 

5932
04:29:05,329 --> 04:29:07,264
BUDDING TYPE PHENOTYPE FROM IT.

5933
04:29:07,264 --> 04:29:08,599
IT WILL BE INTERESTING TO SEE 

5934
04:29:08,599 --> 04:29:10,201
WHAT THE DELETION OF THAT LOOKED

5935
04:29:10,201 --> 04:29:12,636
LIKE TOO, BUT THAT HASN'T BEEN 

5936
04:29:12,636 --> 04:29:20,478
DONE YET.

5937
04:29:20,478 --> 04:29:23,314
INTERESTING NOW, 22Q11.3, WHICH 

5938
04:29:23,314 --> 04:29:25,382
HAS IN THE UPPER RIGHT, THE 

5939
04:29:25,382 --> 04:29:27,218
IDENTICAL AMINO ACID SEQUENCE.

5940
04:29:27,218 --> 04:29:29,286
THIS HAS BEEN PRESERVED FOR 10 

5941
04:29:29,286 --> 04:29:31,989
MILLION YEARS IN OUR ANCESTRAL 

5942
04:29:31,989 --> 04:29:33,157
GENOMES, DIFFERENT FROM ANOTHER 

5943
04:29:33,157 --> 04:29:35,759
AMINO ACID THAT WE DON'T THINK 

5944
04:29:35,759 --> 04:29:38,829
IS MEANINGFUL, ALTHOUGH WE 

5945
04:29:38,829 --> 04:29:40,097
HAVEN'T PROVEN IT YET, BUT 

5946
04:29:40,097 --> 04:29:41,799
THERE'S NO REASON TO BELIEVE 

5947
04:29:41,799 --> 04:29:43,634
THAT POSITION HAS ANY EFFECT ON 

5948
04:29:43,634 --> 04:29:44,034
IT.

5949
04:29:44,034 --> 04:29:44,902
THAT DOES NOT HAVE THIS EFFECT.

5950
04:29:44,902 --> 04:29:46,537
IF YOU LOOK AT THE BOTTOM, YOU 

5951
04:29:46,537 --> 04:29:49,106
CAN SEE IT DOES NOT BLOCK THE 

5952
04:29:49,106 --> 04:29:52,309
BUDDING AND RELEASE.

5953
04:29:52,309 --> 04:29:53,811
SO WHY DOES IT DO THAT?

5954
04:29:53,811 --> 04:29:55,679
THIS IS THE GAG GENE OF THE 

5955
04:29:55,679 --> 04:29:56,680
ANCESTRAL VIRUS.

5956
04:29:56,680 --> 04:29:58,182
YOU LOOK, AND YOU DON'T SEE 

5957
04:29:58,182 --> 04:29:58,816
ANYTHING HAPPENING.

5958
04:29:58,816 --> 04:30:00,384
YOU LOOK AT THE TOP PICTURE, AND

5959
04:30:00,384 --> 04:30:01,585
YOU DON'T SEE ANYTHING HAPPENING

5960
04:30:01,585 --> 04:30:02,753
WITH THE MEMBRANE.

5961
04:30:02,753 --> 04:30:04,455
YOU DO SEE BLOBS AND STUFF 

5962
04:30:04,455 --> 04:30:04,688
INSIDE.

5963
04:30:04,688 --> 04:30:06,924
WHEN WE LOOK AT THE BLOBS OF 

5964
04:30:06,924 --> 04:30:17,434
HIGHER MACK -- MAGNIFY , WHAT 

5965
04:30:19,370 --> 04:30:21,705
YOU SEE, IF YOU REMEMBER YOUR 

5966
04:30:21,705 --> 04:30:23,407
HISTORICAL SEROLOGY, SOMETHING 

5967
04:30:23,407 --> 04:30:25,476
YOU CALL A PARTICLES.

5968
04:30:25,476 --> 04:30:29,647
MANY RETROVIRUSES, INCLUDING 

5969
04:30:29,647 --> 04:30:32,516
MPNV AND OTHERS, DO NOT MATURE 

5970
04:30:32,516 --> 04:30:34,084
THE VIRUS MEMBRANE THE WAY HIV 

5971
04:30:34,084 --> 04:30:35,853
AND MOST OTHER RETROVIRUSES WE 

5972
04:30:35,853 --> 04:30:38,155
WORK WITH DO, THEY MATURE AS 

5973
04:30:38,155 --> 04:30:41,625
INTERCELLULAR PARTICLES, 

5974
04:30:41,625 --> 04:30:43,460
INTERCELLULARPLASMIC PARTICLES 

5975
04:30:43,460 --> 04:30:45,729
THAT THEN FIND THEIR WAY TO THE 

5976
04:30:45,729 --> 04:30:47,164
MEMBRANE AND WRAP THE MEMBRANE 

5977
04:30:47,164 --> 04:30:48,065
AROUND THEM.

5978
04:30:48,065 --> 04:30:49,500
THAT SORT OF SEEMS TO BE WHAT'S 

5979
04:30:49,500 --> 04:30:51,435
HAPPENING HERE, BUT THESE NEVER 

5980
04:30:51,435 --> 04:30:53,103
GET OUT TO THE MEMBRANE, AND 

5981
04:30:53,103 --> 04:30:54,705
THEY'RE INEFFECTIVE IN DOING 

5982
04:30:54,705 --> 04:30:54,905
THAT.

5983
04:30:54,905 --> 04:30:57,575
YOU SEE THAT, THEY SEEM NOT TO 

5984
04:30:57,575 --> 04:30:58,442
CLOSE COMPLETELY EITHER.

5985
04:30:58,442 --> 04:31:01,178
YOU APPROACH THE IDENTICAL AMINO

5986
04:31:01,178 --> 04:31:01,979
ACID SEQUENCE, LIKE THE ONE I 

5987
04:31:01,979 --> 04:31:04,315
SHOWED YOU BEFORE.

5988
04:31:04,315 --> 04:31:06,617
AND THEN NOT AN IMPORTANT ONE, 

5989
04:31:06,617 --> 04:31:08,185
AS FAR AS WE CAN TELL.

5990
04:31:08,185 --> 04:31:11,522
THE DIFFERENCE IS IN THE MATRIX 

5991
04:31:11,522 --> 04:31:14,258
PROTEIN OF THE ANCESTRAL VIRUS.

5992
04:31:14,258 --> 04:31:16,393
MOST MATRIX PROTEINS WE'VE 

5993
04:31:16,393 --> 04:31:19,530
TALKED ABOUT AT THE AMINO 

5994
04:31:19,530 --> 04:31:27,304
TERMINUS HAVE DONE ONLY GLY

5995
04:31:27,304 --> 04:31:28,272
GLYCOLSYLATION SITE, AND THAT 

5996
04:31:28,272 --> 04:31:29,773
ALLOWS THE ASSOCIATION AND THE 

5997
04:31:29,773 --> 04:31:30,741
BUDDING PROCESS TO GO.

5998
04:31:30,741 --> 04:31:35,479
HOWEVER, IF YOU LOOK AT THE 

5999
04:31:35,479 --> 04:31:38,716
LTR5B, IF YOU LOOK, THERE ARE 

6000
04:31:38,716 --> 04:31:40,250
MORE ACIDIC AMINO ACIDS IN THE 

6001
04:31:40,250 --> 04:31:41,585
REGION THAN BASIC AMINO ACIDS.

6002
04:31:41,585 --> 04:31:44,154
AND THE PREDICTED PK OF THIS IS 

6003
04:31:44,154 --> 04:31:45,456
ACTUALLY QUITE ACIDIC.

6004
04:31:45,456 --> 04:31:47,124
THERE'S NO WAY THIS PROTEIN IS 

6005
04:31:47,124 --> 04:31:48,392
GOING TO ASSOCIATE WITH THE 

6006
04:31:48,392 --> 04:31:49,827
MEMBRANE THE SAME WAY.

6007
04:31:49,827 --> 04:31:50,527
THAT'S WHAT HAPPENS.

6008
04:31:50,527 --> 04:31:51,862
THESE DON'T GO TO THE MEMBRANE 

6009
04:31:51,862 --> 04:31:52,463
THEY ASSOCIATE.

6010
04:31:52,463 --> 04:31:53,631
THEY DON'T GO TO THE MEMBRANE AT

6011
04:31:53,631 --> 04:31:54,765
THAT POINT.

6012
04:31:54,765 --> 04:31:55,966
HOW THEY DO GET THERE EVENTUALLY

6013
04:31:55,966 --> 04:31:57,801
IN THE REAL VIRUS, I HAVE NO 

6014
04:31:57,801 --> 04:31:58,669
IDEA, BUT THEY DON'T ASSOCIATE 

6015
04:31:58,669 --> 04:32:05,042
WITH IT THERE.

6016
04:32:05,042 --> 04:32:06,410
I'LL TELL YOU HOW WE INTERPRET 

6017
04:32:06,410 --> 04:32:07,878
ALL THIS IN EVOLUTIONARY TERMS 

6018
04:32:07,878 --> 04:32:09,780
AT THE END, BUT FIRST I WANT TO 

6019
04:32:09,780 --> 04:32:10,681
POINT OUT SOMETHING ELSE.

6020
04:32:10,681 --> 04:32:12,783
IF YOU LOOK AT THE TWO LITTLE 

6021
04:32:12,783 --> 04:32:15,819
SETS OF BARS, IF YOU DO THE SAME

6022
04:32:15,819 --> 04:32:18,789
KINDS OF EXPERIMENTS WITHIN MLV,

6023
04:32:18,789 --> 04:32:22,493
THE GAMMA RETROVIRUS, OR THE 

6024
04:32:22,493 --> 04:32:24,928
ALPHA RETROVIRUS, THIS HAS NO 

6025
04:32:24,928 --> 04:32:27,731
EFFECT ON THESE RETROVIRUSES, 

6026
04:32:27,731 --> 04:32:30,334
WHICH MAKES SENSE BECAUSE 

6027
04:32:30,334 --> 04:32:31,301
COEXISTENCE WOULD BE NECESSARY 

6028
04:32:31,301 --> 04:32:32,770
FOR THIS EFFECT TO OCCUR.

6029
04:32:32,770 --> 04:32:35,739
IF YOU LOOK AT MPMV, THIS IS A 

6030
04:32:35,739 --> 04:32:39,176
VIRUS THAT ALSO ACCESSIBLES 

6031
04:32:39,176 --> 04:32:39,877
INTRACELLULAR, YOU CAN SEE THIS 

6032
04:32:39,877 --> 04:32:46,884
IS INHIBITED BY THE 22Q11.23, 

6033
04:32:46,884 --> 04:32:49,887
WHICH HAS THE INTERPLASMIC 

6034
04:32:49,887 --> 04:32:50,487
ASSEMBLY AS WELL.

6035
04:32:50,487 --> 04:32:52,556
THIS IS THE SAME KIND OF 

6036
04:32:52,556 --> 04:32:58,696
COASSEMBLY DEFECT, JUST IN 

6037
04:32:58,696 --> 04:33:02,866
INTERCYTOPLASMIC.

6038
04:33:02,866 --> 04:33:03,834
REMARKABLY, AND WHY I WAS ABLE 

6039
04:33:03,834 --> 04:33:06,503
TO GET A GRANT ON THIS SUBJECT, 

6040
04:33:06,503 --> 04:33:11,141
THE -- THIS PROTEIN DOES BLOCK 

6041
04:33:11,141 --> 04:33:16,980
THE 3Q12.23 PROTEIN DOES BLOCK 

6042
04:33:16,980 --> 04:33:20,017
THE HIV ASSEMBLY.

6043
04:33:20,017 --> 04:33:20,984
AGAIN, YOU SEE IT'S ALMOST 

6044
04:33:20,984 --> 04:33:22,619
EXACTLY THE SAME KIND OF CURVE I

6045
04:33:22,619 --> 04:33:30,661
SHOWED YOU BEFORE WHEN YOU 

6046
04:33:30,661 --> 04:33:32,863
TITRATE IT AGAINST HIV, AND THE 

6047
04:33:32,863 --> 04:33:34,698
LTR RELEASE.

6048
04:33:34,698 --> 04:33:38,302
SO IT BLOCKS HIV PARTICLE 

6049
04:33:38,302 --> 04:33:40,037
MATURATION WHEN DOSED IN THE 

6050
04:33:40,037 --> 04:33:40,971
INTENDED MANNER.

6051
04:33:40,971 --> 04:33:42,873
IF YOU LOOK AT PICTURES -- 

6052
04:33:42,873 --> 04:33:45,542
HERE'S HIV ALONE ON THE LEFT.

6053
04:33:45,542 --> 04:33:48,512
HERE'S THE GAG PROFILE, SO IT 

6054
04:33:48,512 --> 04:33:49,980
GETS MATURE PARTICLES.

6055
04:33:49,980 --> 04:33:52,616
IF YOU ADD THAT WITH KCON GAG, 

6056
04:33:52,616 --> 04:33:53,984
WHICH ACTUALLY HAS BEEN SHOWN BY

6057
04:33:53,984 --> 04:33:56,620
OTHERS, INCLUDING A PAPER WITH 

6058
04:33:56,620 --> 04:33:58,355
ALAN REIN, TO BE SOMEWHAT 

6059
04:33:58,355 --> 04:34:00,724
INHIBITORY, BUT NOWHERE NEARLY 

6060
04:34:00,724 --> 04:34:05,763
AS MUCH AS 3Q12.3.

6061
04:34:05,763 --> 04:34:07,531
YOU GET PERFECTLY GOOD LOOKING 

6062
04:34:07,531 --> 04:34:08,799
STRUCTURES, BUT YOU GET THE SAME

6063
04:34:08,799 --> 04:34:10,400
KIND OF ASSEMBLY DEFECTS ON THE 

6064
04:34:10,400 --> 04:34:12,903
RIGHT THAT WE SAW WITH THE WILD 

6065
04:34:12,903 --> 04:34:13,604
TY

6066
04:34:13,604 --> 04:34:18,742
TYPE.

6067
04:34:18,742 --> 04:34:22,412
HERV-K.

6068
04:34:22,412 --> 04:34:23,847
SO IN ALMOST ALL THE EXPERIMENTS

6069
04:34:23,847 --> 04:34:26,416
I SHOWED YOU BEFORE, EXCEPT FOR 

6070
04:34:26,416 --> 04:34:33,724
THE END, SET UP A NANO-BRET 

6071
04:34:33,724 --> 04:34:44,268
ASSAY, WHICH USES BIOLEUKEMIA --

6072
04:34:45,068 --> 04:34:46,904
BIOLUMINESCENCE ASSAY.

6073
04:34:46,904 --> 04:34:50,040
SO YOU HAVE TWO POTENTIAL 

6074
04:34:50,040 --> 04:34:54,578
INTERACTORS, ONE HAS THE 

6075
04:34:54,578 --> 04:34:56,280
NANO-LUKE TAG AND THE OTHER HAS 

6076
04:34:56,280 --> 04:34:57,347
A HALO TAG.

6077
04:34:57,347 --> 04:35:00,717
THE EMISSIONS FROM THE NANO-LUKE

6078
04:35:00,717 --> 04:35:02,820
CAN THEN STIMULATE THE HALO AND 

6079
04:35:02,820 --> 04:35:05,455
GIVE THE HALO SPECTRAL 

6080
04:35:05,455 --> 04:35:06,890
FLUORESCENCE, AND YOU CAN USE 

6081
04:35:06,890 --> 04:35:08,125
THAT AS AN INTRACELLULAR MEASURE

6082
04:35:08,125 --> 04:35:10,561
OF INTERACTION, AND YOU CAN SEE 

6083
04:35:10,561 --> 04:35:16,366
FROM THE BARS THAT THE HALO 

6084
04:35:16,366 --> 04:35:19,903
ALONE DOES NOTHING OF 

6085
04:35:19,903 --> 04:35:20,637
SIGNIFIC

6086
04:35:20,637 --> 04:35:20,971
SIGNIFICANCE.

6087
04:35:20,971 --> 04:35:22,706
THEN ON THE RIGHT, THE OTHER 

6088
04:35:22,706 --> 04:35:26,643
CONTROLS, MLV AND MPNV DON'T 

6089
04:35:26,643 --> 04:35:28,779
HAVE ANY EFFECT, SHOW NO 

6090
04:35:28,779 --> 04:35:32,349
INTERACTIONS, WHEREAS BOTH THE 

6091
04:35:32,349 --> 04:35:36,053
3Q12.3 GAG HALO, THE KCON GAG 

6092
04:35:36,053 --> 04:35:38,355
HALO INTERACTS WITH ITSELF, AS 

6093
04:35:38,355 --> 04:35:39,289
YOU'D EXPECT.

6094
04:35:39,289 --> 04:35:45,062
THE 3Q12.3 HALO INTERACTS, AND 

6095
04:35:45,062 --> 04:35:46,263
HIV INTERACTS WITH MORE WHAT 

6096
04:35:46,263 --> 04:35:48,332
SEEM TO BE THE SAME LEVEL.

6097
04:35:48,332 --> 04:35:50,300
THE FACT THAT THEY INTERACT WITH

6098
04:35:50,300 --> 04:35:53,437
QUITE SURPRISING, THESE TWO 

6099
04:35:53,437 --> 04:35:54,605
CAPSID PROTEINS HAVE VERY LITTLE

6100
04:35:54,605 --> 04:35:56,440
SIMILARITY IN SEQUENCE, BUT THEY

6101
04:35:56,440 --> 04:35:58,642
ARE QUITE SIMILAR IN STRUCTURE 

6102
04:35:58,642 --> 04:36:01,411
AT THE CRITICAL JUNCTION WHERE 

6103
04:36:01,411 --> 04:36:06,583
THE HEXAMER IS FORMED IN THE 

6104
04:36:06,583 --> 04:36:07,451
IMMATURE FORM.

6105
04:36:07,451 --> 04:36:08,552
YOU CAN SEE THE GRAY TRACE ON 

6106
04:36:08,552 --> 04:36:10,654
THE RIGHT IS HIV.

6107
04:36:10,654 --> 04:36:19,096
THE ORANGE IS HML-2.

6108
04:36:19,096 --> 04:36:21,365
THEY'RE REMARKABLY SIMILAR IN 

6109
04:36:21,365 --> 04:36:23,100
THEIR STRUCTURE EVEN THOUGH THE 

6110
04:36:23,100 --> 04:36:24,134
AMINO ACID SEQUENCE IS 

6111
04:36:24,134 --> 04:36:24,401
DIFFERENT.

6112
04:36:24,401 --> 04:36:26,536
WE IMAGINE THIS DOES ALLOW AT 

6113
04:36:26,536 --> 04:36:29,640
LEAST SOME COASSEMBLY OF THE 

6114
04:36:29,640 --> 04:36:32,209
TWO, AND THAT IS WHAT ALLOWS IT 

6115
04:36:32,209 --> 04:36:36,513
TO BE PROTECTIVE, IF YOU LIKE, 

6116
04:36:36,513 --> 04:36:39,116
EVEN THOUGH, IF YOU LOOK AT THE 

6117
04:36:39,116 --> 04:36:40,851
SEQUENCE OF THESE, AS SHOWN BY 

6118
04:36:40,851 --> 04:36:44,054
THE TREE THERE, THERE'S REALLY 

6119
04:36:44,054 --> 04:36:49,159
NO PARTICULAR SPECIAL SEQUENCE 

6120
04:36:49,159 --> 04:36:50,627
RELATIONSHIP BETWEEN THESE 

6121
04:36:50,627 --> 04:36:51,895
PROTEINS OVERALL.

6122
04:36:51,895 --> 04:36:53,964
NOW TO THE SORT OF BOTTOM LINE 

6123
04:36:53,964 --> 04:36:54,164
HERE.

6124
04:36:54,164 --> 04:36:56,099
WE INFERRED FROM ALL OF THIS 

6125
04:36:56,099 --> 04:36:58,835
WITH A LITTLE BIT OF SPECULATION

6126
04:36:58,835 --> 04:37:02,306
MIXED IN AN EVOLUTIONARY PATHWAY

6127
04:37:02,306 --> 04:37:05,108
BY WHICH THIS HAS COME TO BE.

6128
04:37:05,108 --> 04:37:12,449
YOU START OUT WITH THE LTR-5B, 

6129
04:37:12,449 --> 04:37:15,252
THE ANCESTRAL VIRUS, TO THE HS 

6130
04:37:15,252 --> 04:37:17,788
GROUP, AND REPLICATED IN OUR 

6131
04:37:17,788 --> 04:37:19,156
ANCESTORS GOING BACK 25 MILLION 

6132
04:37:19,156 --> 04:37:19,356
YEARS.

6133
04:37:19,356 --> 04:37:20,991
I ONLY SHOW WHAT'S HAPPENED IN 

6134
04:37:20,991 --> 04:37:23,260
THE LAST 10 MILLION YEARS.

6135
04:37:23,260 --> 04:37:26,730
AT SOME POINT, AND ALSO WE THINK

6136
04:37:26,730 --> 04:37:28,999
IN THE HUMAN, COMMON ANCESTORS 

6137
04:37:28,999 --> 04:37:32,536
HUMANS AND GORILLAS, BUT PRIOR 

6138
04:37:32,536 --> 04:37:39,176
TO THE OTHER ONE, THIS MUTATION 

6139
04:37:39,176 --> 04:37:41,645
INSERTED, WE SPECULATE THAT THIS

6140
04:37:41,645 --> 04:37:43,413
WAS THEN PROTECTED AGAINST 

6141
04:37:43,413 --> 04:37:45,916
REPLICATION OF THE 5B TYPE 

6142
04:37:45,916 --> 04:37:48,318
VIRUSES WHICH HAD THIS 

6143
04:37:48,318 --> 04:37:50,420
INTRACELLULAR PLASMIC ASSEMBLY.

6144
04:37:50,420 --> 04:37:54,358
PERHAPS IN RESPONSE TO THIS, THE

6145
04:37:54,358 --> 04:37:57,828
5HS TYPE, WHICH HAS THE 

6146
04:37:57,828 --> 04:37:59,930
CYTOPLASMIC ASSEMBLY MAY HAVE 

6147
04:37:59,930 --> 04:38:01,932
EVOLVED BY MUTATIONS OF 

6148
04:38:01,932 --> 04:38:03,433
DUPLICATION, AS YOU CAN SEE IN 

6149
04:38:03,433 --> 04:38:05,235
THE PREVIOUS SLIDE, IN THE 

6150
04:38:05,235 --> 04:38:07,971
MATRIX N-TERMINAL REGION TO GIVE

6151
04:38:07,971 --> 04:38:11,541
IT MEMBRANE ASSOCIATED ASSEMBLY,

6152
04:38:11,541 --> 04:38:13,043
AND THAT HAS ALLOWED IT TO 

6153
04:38:13,043 --> 04:38:15,979
ESCAPE FROM THE EFFECTS OF THIS 

6154
04:38:15,979 --> 04:38:16,580
INHIBITORY EFFECT MIGHT HAVE 

6155
04:38:16,580 --> 04:38:18,515
BEEN.

6156
04:38:18,515 --> 04:38:19,683
THAT WAS THEN IN THE COURSE OF 

6157
04:38:19,683 --> 04:38:22,219
THE REPLICATION THAT VIRUS, THE 

6158
04:38:22,219 --> 04:38:23,820
RECOMBINATION OCCURRED, WHICH 

6159
04:38:23,820 --> 04:38:25,655
BROUGHT IN THE CAPSID, 

6160
04:38:25,655 --> 04:38:27,791
PARTICULARLY THE CAPSID DOMAIN 

6161
04:38:27,791 --> 04:38:33,096
FROM THE RESIDENT 5B VIRUS.

6162
04:38:33,096 --> 04:38:35,932
THAT THEN REPLICATED ALL THE WAY

6163
04:38:35,932 --> 04:38:38,468
UP AND PROBABLY ALSO GAVE AT 

6164
04:38:38,468 --> 04:38:40,404
LEAST SOME PROTECTION INTO 

6165
04:38:40,404 --> 04:38:42,305
HUMANS, AND THEN AT SOME POINT 

6166
04:38:42,305 --> 04:38:43,874
IN THE HUMAN SPECIFIC ANCESTRY 

6167
04:38:43,874 --> 04:38:46,176
IN THE LAST 5 MILLION YEARS OR 

6168
04:38:46,176 --> 04:38:48,311
SO, THAT DOES DUPLICATION IN THE

6169
04:38:48,311 --> 04:38:50,547
LTR OCCURRED, AND THAT GAVE IT A

6170
04:38:50,547 --> 04:38:54,117
MUCH BETTER EXPRESSION.

6171
04:38:54,117 --> 04:38:55,685
AND WE SPECULATE IT MAY HAVE 

6172
04:38:55,685 --> 04:38:56,686
CONTRIBUTED AT LEAST TO THE 

6173
04:38:56,686 --> 04:38:58,555
EXTINCTION OF THAT AS A VIRUS.

6174
04:38:58,555 --> 04:39:01,224
THAT IS OUR SORT OF CURRENT 

6175
04:39:01,224 --> 04:39:02,426
HYPOTHETICAL FRAMEWORK FOR 

6176
04:39:02,426 --> 04:39:04,528
THINKING ABOUT HOW THE EVOLUTION

6177
04:39:04,528 --> 04:39:06,296
OF THIS VIRUS, THIS PARTICULAR 

6178
04:39:06,296 --> 04:39:07,564
VIRUS AND RESTRICTION FACTOR 

6179
04:39:07,564 --> 04:39:09,332
MIGHT HAVE OCCURRED.

6180
04:39:09,332 --> 04:39:10,500
MANY PEOPLE IN THE LAB HAVE MADE

6181
04:39:10,500 --> 04:39:12,536
THIS POSSIBLE.

6182
04:39:12,536 --> 04:39:14,171
THIS IS A LONG SERIES OF 

6183
04:39:14,171 --> 04:39:15,972
STUDENTS AND POST DOCS WHO 

6184
04:39:15,972 --> 04:39:19,676
WORKED ON THIS OVER THE YEARS, 

6185
04:39:19,676 --> 04:39:26,750
RAVI, MEAGHAN, FARRAH, AIDAN, 

6186
04:39:26,750 --> 04:39:28,285
AND MOST OF THEM, ALL THE WORK I

6187
04:39:28,285 --> 04:39:30,487
SHOWED YOU, IS NOW THE WORK OF 

6188
04:39:30,487 --> 04:39:32,189
MIKE FREEMAN, NOW A POST DOC IN 

6189
04:39:32,189 --> 04:39:34,691
THE LAB, WITH THE ENORMOUS 

6190
04:39:34,691 --> 04:39:38,628
SUPPORT TO GET THE BEAUTIFUL 

6191
04:39:38,628 --> 04:39:40,097
ELECTROMICROGRAPHS.

6192
04:39:40,097 --> 04:39:42,099
WITH PARTICULAR THANKS OVERALL 

6193
04:39:42,099 --> 04:39:45,202
TO ALAN REIN, WHO EVER SINCE HE 

6194
04:39:45,202 --> 04:39:48,004
JOINED THE DRP BACK IN '98 OR SO

6195
04:39:48,004 --> 04:39:49,739
HAS BEEN A REALLY VALUED 

6196
04:39:49,739 --> 04:39:51,808
COLLEAGUE AND SOURCE OF 

6197
04:39:51,808 --> 04:39:54,478
INSPIRATION ON CRITICAL SCIENCE.

6198
04:39:54,478 --> 04:40:04,754
I'LL THANK YOU.

6199
04:40:07,390 --> 04:40:08,525
>> YOU'VE SHOWN THIS DATA.

6200
04:40:08,525 --> 04:40:09,593
I DON'T REMEMBER THE RESULTS.

6201
04:40:09,593 --> 04:40:12,362
HOW BROAD IS THE CELLULAR TISSUE

6202
04:40:12,362 --> 04:40:13,797
DISTRIBUTION OF THE INTERFERING 

6203
04:40:13,797 --> 04:40:14,097
RETROVIRUS?

6204
04:40:14,097 --> 04:40:15,232
>> IT'S EVERYWHERE.

6205
04:40:15,232 --> 04:40:16,766
THE ONLY PLACE IN THE DATA SET 

6206
04:40:16,766 --> 04:40:19,636
IT DIDN'T SHOW UP WAS IN WHOLE 

6207
04:40:19,636 --> 04:40:21,805
BLOOD, WHICH YOU WOULD ARGUE 

6208
04:40:21,805 --> 04:40:24,107
COULD WELL BE ALLOWING -- IF IT 

6209
04:40:24,107 --> 04:40:26,109
HAD BEEN EXPRESSED THERE WELL 

6210
04:40:26,109 --> 04:40:27,711
ENOUGH, WE MIGHT NEVER HAVE HAD 

6211
04:40:27,711 --> 04:40:28,445
THE HIV PANDEMIC.

6212
04:40:28,445 --> 04:40:31,114
NOW WE'RE GETTING REALLY 

6213
04:40:31,114 --> 04:40:33,316
SPECULATIVE OF COURSE.

6214
04:40:33,316 --> 04:40:35,519
IN FACT, MY -- ONE OF THE 

6215
04:40:35,519 --> 04:40:37,220
STRETCH AIMS OF MY GRANT IS TO 

6216
04:40:37,220 --> 04:40:39,189
TRY TO GET IT EXPRESSED AGAIN TO

6217
04:40:39,189 --> 04:40:40,390
SEE IF WE CAN ACTUALLY SEE WHAT 

6218
04:40:40,390 --> 04:40:47,230
THE EFFECT WOULD BE.

6219
04:40:47,230 --> 04:40:51,501
>> HOW CAN THE ASSAY WITH
THE 

6220
04:40:51,501 --> 04:40:52,836
CYTOPLASMIC PROTECTION DESCRIBED

6221
04:40:52,836 --> 04:40:54,137
BY ERIC HUNTER SEVERAL YEARS 

6222
04:40:54,137 --> 04:40:57,407
AGO, AND IT'S FURTHER DOWN -- 

6223
04:40:57,407 --> 04:40:59,476
>> I WOULD HAVE TO DO
OTHER 

6224
04:40:59,476 --> 04:41:01,845
THINGS TO MPNV TO MAKE IT 

6225
04:41:01,845 --> 04:41:02,112
NECESSARY.

6226
04:41:02,112 --> 04:41:03,413
>> ARE THERE CHANGES IN
THAT 

6227
04:41:03,413 --> 04:41:03,647
REGION?

6228
04:41:03,647 --> 04:41:05,148
>> TO BE HONEST WITH YOU,
WE 

6229
04:41:05,148 --> 04:41:05,682
HAVEN'T LOOKED.

6230
04:41:05,682 --> 04:41:06,783
THAT'S A VERY GOOD POINT.

6231
04:41:06,783 --> 04:41:16,860
WE SHOULD TAKE A LOOK AT THAT.

6232
04:41:16,860 --> 04:41:23,400
>> WITH THE MYRISILATION
-- 

6233
04:41:23,400 --> 04:41:24,634
>> WE HAVEN'T LOOKED TO
SEE, 

6234
04:41:24,634 --> 04:41:26,069
BUT THE SIGNAL IS THERE.

6235
04:41:26,069 --> 04:41:36,413
THANK YOU VERY MUCH.

6236
04:41:53,096 --> 04:41:54,931
>> OUR NEXT SPEAKER HAS A
TRULY 

6237
04:41:54,931 --> 04:41:55,832
UNIQUE DISTINCTION.

6238
04:41:55,832 --> 04:41:57,767
HE HAS BEEN IN THE FIELD LONGER 

6239
04:41:57,767 --> 04:41:59,135
THAN I HAVE.

6240
04:41:59,135 --> 04:42:03,373
THE REASON I KNOW THAT IS WHEN I

6241
04:42:03,373 --> 04:42:06,176
JOINED RUBIN'S LAB IN THE SUMMER

6242
04:42:06,176 --> 04:42:11,881
OF 1962, PETER WAS JUST LEAVING.

6243
04:42:11,881 --> 04:42:15,018
WE OVERLAPPED FOR PROBABLY A FEW

6244
04:42:15,018 --> 04:42:17,721
WEEKS IN HARRY RUBIN'S LAB WHEN 

6245
04:42:17,721 --> 04:42:20,323
HE WAS JUST LEAVING TO FORM HIS 

6246
04:42:20,323 --> 04:42:22,792
OWN LAB.

6247
04:42:22,792 --> 04:42:26,830
SO IT'S BEEN MANY, MANY DECADES,

6248
04:42:26,830 --> 04:42:31,468
AND HE HAS PERFORMED, MADE MANY 

6249
04:42:31,468 --> 04:42:37,173
CRUCIAL DISCOVERIES PURSUING THE

6250
04:42:37,173 --> 04:42:40,277
VIROLOGY OF THE ALPHA 

6251
04:42:40,277 --> 04:42:42,712
RETROVIRUSES AND THEN ACTUALLY 

6252
04:42:42,712 --> 04:42:45,248
HE WAS A PIONEER IN GOING AROUND

6253
04:42:45,248 --> 04:42:47,817
AND DIGGING VIRUSES OUT OF 

6254
04:42:47,817 --> 04:42:48,885
PEOPLE'S FREEZERS AND 

6255
04:42:48,885 --> 04:42:52,756
DISCOVERING NEW ONCOGENES THAT 

6256
04:42:52,756 --> 04:42:52,922
WAY.

6257
04:42:52,922 --> 04:42:56,192
IN MORE RECENT YEARS, I THINK 

6258
04:42:56,192 --> 04:43:00,130
HE'S SPECIALIZED IN LIPID 

6259
04:43:00,130 --> 04:43:04,434
SIGNALING, WHICH REALLY ARISES 

6260
04:43:04,434 --> 04:43:08,805
FROM HIS DISCOVERY OF YET 

6261
04:43:08,805 --> 04:43:12,042
UNEXPECTED NEW ONCOGENES AND 

6262
04:43:12,042 --> 04:43:16,379
CONSIDERING THE POSSIBLE 

6263
04:43:16,379 --> 04:43:18,648
APPLICATIONS OF HIS DISCOVERIES 

6264
04:43:18,648 --> 04:43:23,586
ON LIPID SIGNALING IN CANCER.

6265
04:43:23,586 --> 04:43:24,487
WITHOUT FURTHER ADO HERE, 

6266
04:43:24,487 --> 04:43:28,758
WELCOME TO THE MEETING.

6267
04:43:28,758 --> 04:43:29,926
>> THANK YOU FOR YOUR KIND
AND 

6268
04:43:29,926 --> 04:43:32,295
GENEROUS INTRODUCTION.

6269
04:43:32,295 --> 04:43:34,764
I AM HONORED AND DELIGHTED TO 

6270
04:43:34,764 --> 04:43:38,268
SPEAK AT THE SYMPOSIUM 

6271
04:43:38,268 --> 04:43:40,704
CELEBRATING ALAN REIN'S WORK.

6272
04:43:40,704 --> 04:43:42,472
ALAN AND I SHARE EARLY 

6273
04:43:42,472 --> 04:43:45,075
SCIENTIFIC ROOTS, AS YOU JUST 

6274
04:43:45,075 --> 04:43:45,308
LEARNED.

6275
04:43:45,308 --> 04:43:50,747
WE WERE BOTH WORKING ON THE 

6276
04:43:50,747 --> 04:43:51,681
NEUROSARCOMA VIRUS IN THE SAME 

6277
04:43:51,681 --> 04:43:53,650
LAB IN BERKELEY.

6278
04:43:53,650 --> 04:43:55,885
IN THIS SYMPOSIUM I WILL BE THE 

6279
04:43:55,885 --> 04:43:58,288
ONLY SPEAKER TALKING MAINLY 

6280
04:43:58,288 --> 04:44:02,659
ABOUT AVIAN VIRUSES, AND AS THE 

6281
04:44:02,659 --> 04:44:04,427
LAST SPEAKER, I HAVE DECIDED TO 

6282
04:44:04,427 --> 04:44:07,597
TAKE A LIGHT HEARTED APPROACH 

6283
04:44:07,597 --> 04:44:11,401
AND DISTILL IT INTO SEVEN 

6284
04:44:11,401 --> 04:44:13,803
SIMPLIFIED LESSONS LEARNED 

6285
04:44:13,803 --> 04:44:15,238
DURING A LIFELONG SCIENTIFIC 

6286
04:44:15,238 --> 04:44:21,611
EDUCATION.

6287
04:44:21,611 --> 04:44:24,114
I NEED TO EMPHASIZE THESE ARE 

6288
04:44:24,114 --> 04:44:25,014
LESSONS LEARNED.

6289
04:44:25,014 --> 04:44:27,384
THEY ARE NOT MEANT AS CAREER 

6290
04:44:27,384 --> 04:44:27,684
ADVICE.

6291
04:44:27,684 --> 04:44:38,228
SO LET ME BEGIN SCREEN SHARING.

6292
04:45:02,485 --> 04:45:03,787
LESSON NUMBER ONE, YOU SHOULD 

6293
04:45:03,787 --> 04:45:05,622
JOIN AN EMERGING, NOVEL FIELD, 

6294
04:45:05,622 --> 04:45:08,091
AND APPLY NEW CONCEPTS AND 

6295
04:45:08,091 --> 04:45:09,726
GENERATE NEW TECHNOLOGIES.

6296
04:45:09,726 --> 04:45:11,060
DON'T JOIN A MATURE FIELD.

6297
04:45:11,060 --> 04:45:12,996
I THINK ALL OF YOU HAVE APPLIED 

6298
04:45:12,996 --> 04:45:19,269
THAT LESSON IN YOUR CAREERS WHEN

6299
04:45:19,269 --> 04:45:22,005
YOU JOINED HIV.

6300
04:45:22,005 --> 04:45:25,341
THE EMERGING FIELD OF MY TIME 

6301
04:45:25,341 --> 04:45:28,278
WAS A QUANTITATIVE ANALYSIS FOR 

6302
04:45:28,278 --> 04:45:30,980
TUMOR VIRUS REPLICATION AND A 

6303
04:45:30,980 --> 04:45:31,915
VIRUS-INDUCED CELLULAR 

6304
04:45:31,915 --> 04:45:32,282
TRANSFORMATION.

6305
04:45:32,282 --> 04:45:37,654
THAT IS WORK THAT START ED AT CL

6306
04:45:37,654 --> 04:45:45,528
TECH WITH HARRY RUBIN AND HOWARD

6307
04:45:45,528 --> 04:45:45,762
TENEMAN.

6308
04:45:45,762 --> 04:45:47,864
AND THE BASIC TECHNOLOGY WAS THE

6309
04:45:47,864 --> 04:45:52,035
FOCUS ASSAY OF ROUS SARCOMA 

6310
04:45:52,035 --> 04:45:52,235
VIRUS.

6311
04:45:52,235 --> 04:45:56,306
THE FIRST ASSAY THAT ALLOWED YOU

6312
04:45:56,306 --> 04:45:59,142
TO QUANTIFY ONCOGENIC 

6313
04:45:59,142 --> 04:45:59,609
TRANSFORMATION.

6314
04:45:59,609 --> 04:46:02,879
HERE YOU SEE AN ILLUSTRATION OF 

6315
04:46:02,879 --> 04:46:05,648
THIS NORMAL AVIAN FIBROBLAST TO 

6316
04:46:05,648 --> 04:46:09,986
THE LEFT AND ROUS SARCOMA VIRUS 

6317
04:46:09,986 --> 04:46:11,688
TRANSFORMED FIBROBLASTS TO THE 

6318
04:46:11,688 --> 04:46:11,888
RIGHT.

6319
04:46:11,888 --> 04:46:14,657
YOU COULD REALLY HAVE THIS 

6320
04:46:14,657 --> 04:46:18,428
TRANSFORMATION IN VERY DISCRETE 

6321
04:46:18,428 --> 04:46:20,730
FOCI, AND THE FOCI, A SINGLE 

6322
04:46:20,730 --> 04:46:22,398
FOCUS WAS THE RESULT OF 

6323
04:46:22,398 --> 04:46:25,134
INFECTION WITH A SINGLE VIRAL 

6324
04:46:25,134 --> 04:46:25,401
PARTICLE.

6325
04:46:25,401 --> 04:46:26,536
SO IT WAS QUANTITATIVE FROM THE 

6326
04:46:26,536 --> 04:46:36,646
START.

6327
04:46:37,714 --> 04:46:39,082
IT WAS BROUGHT ON BY THE WORK 

6328
04:46:39,082 --> 04:46:41,217
BEING DONE AT CAL TECH.

6329
04:46:41,217 --> 04:46:43,353
LESSON NUMBER 2, TRUST YOUR 

6330
04:46:43,353 --> 04:46:43,620
INSTINCTS.

6331
04:46:43,620 --> 04:46:45,688
CHICKEN TUMOR VIRUSES ARE 

6332
04:46:45,688 --> 04:46:47,223
SIGNIFICANT FOR HUMAN MEDICINE.

6333
04:46:47,223 --> 04:46:49,459
THERE WAS ALWAYS THIS TENDENCY 

6334
04:46:49,459 --> 04:46:53,062
OF DOWNPLAYING THE AVIAN VIRUSES

6335
04:46:53,062 --> 04:46:55,231
BECAUSE THEY COULDN'T POSSIBLY 

6336
04:46:55,231 --> 04:47:05,675
BE RELEVANT FOR HUMANS IN 

6337
04:47:11,915 --> 04:47:12,115
MEDICINE.

6338
04:47:12,115 --> 04:47:13,650
SO THE FIRST IMPORTANT FEATURE 

6339
04:47:13,650 --> 04:47:16,286
THAT SETS AVIAN TUMOR VIRUSES 

6340
04:47:16,286 --> 04:47:19,822
APART IS THAT MOST ROUS SARCOMA 

6341
04:47:19,822 --> 04:47:22,258
VIRUSES ARE NON-DEFECTIVE.

6342
04:47:22,258 --> 04:47:24,127
THEY CONTAIN ALL THE VIRAL GENES

6343
04:47:24,127 --> 04:47:25,828
PLUS THE ONCOGENE.

6344
04:47:25,828 --> 04:47:28,498
THAT'S THE ONLY WAY YOU CAN DO 

6345
04:47:28,498 --> 04:47:39,042
GENETICS IN A CONVENTIONAL WAY.

6346
04:47:39,309 --> 04:47:42,645
AND ONLY AFTER THE ROUS SARCOMA 

6347
04:47:42,645 --> 04:47:45,515
VIRUS WAS FOUND TO BE 

6348
04:47:45,515 --> 04:47:49,485
NON-DEFECTIVE WAS A GENETIC 

6349
04:47:49,485 --> 04:47:51,120
APPROACH TO THIS GENOME 

6350
04:47:51,120 --> 04:47:51,621
POSSIBLE.

6351
04:47:51,621 --> 04:47:53,156
AND THE SECOND ADVANTAGE IS 

6352
04:47:53,156 --> 04:47:56,926
AVIAN RETROVIRUSES USE TEN 

6353
04:47:56,926 --> 04:47:58,962
DIFFERENT SURFACE GLYCOPROTEINS,

6354
04:47:58,962 --> 04:48:02,098
AND IT GIVES YOU ENORMOUS 

6355
04:48:02,098 --> 04:48:04,267
RICHNESS AND WEALTH OF ALL KINDS

6356
04:48:04,267 --> 04:48:09,806
OF TOOLS AND APPROACHES.

6357
04:48:09,806 --> 04:48:12,575
EACH GLYCOPROTEIN TARGETS A 

6358
04:48:12,575 --> 04:48:16,579
SPECIFIC CELL SURFACE RECEPTOR, 

6359
04:48:16,579 --> 04:48:18,581
AND WHEN THE GLYCOPROTEIN 

6360
04:48:18,581 --> 04:48:21,317
OCCUPIES THAT RECEPTOR, THAT 

6361
04:48:21,317 --> 04:48:24,220
INDUCES VIRAL INTERFERENCE.

6362
04:48:24,220 --> 04:48:27,690
LESSON 3 IS TAKE THESE 

6363
04:48:27,690 --> 04:48:32,328
OBSERVATIONS AND BUILD NEW TO

6364
04:48:32,328 --> 04:48:35,164
TOOLS, AND FOR THE AVIAN VIRUSES

6365
04:48:35,164 --> 04:48:39,502
IT WAS CLEAR THAT GLYCOPROTEIN 

6366
04:48:39,502 --> 04:48:42,138
ALLOWED YOU TO CLASSIFY THE 

6367
04:48:42,138 --> 04:48:52,548
VIRUSES INTO SUBGROUPS.

6368
04:48:52,849 --> 04:48:54,717
THAT WAS COMPLEMENTED BY FINDING

6369
04:48:54,717 --> 04:48:56,152
CHICKEN LINES THAT ARE RESISTANT

6370
04:48:56,152 --> 04:48:58,655
TO INFECTION BY A SINGLE VIRAL 

6371
04:48:58,655 --> 04:48:59,922
SUBGROUP BECAUSE THEY LACK THAT 

6372
04:48:59,922 --> 04:49:02,425
PARTICULAR RECEPTOR, AND THIS 

6373
04:49:02,425 --> 04:49:06,162
INTERPLAY OF VIRAL GLYCOPROTEIN 

6374
04:49:06,162 --> 04:49:10,166
AND CELLULAR RECEPTOR COULD BE 

6375
04:49:10,166 --> 04:49:11,901
MADE INTO AN ASSAY OF VIRAL 

6376
04:49:11,901 --> 04:49:14,170
INTERFERENCE.

6377
04:49:14,170 --> 04:49:16,973
SO YOU HAVE TO REMEMBER THIS IS 

6378
04:49:16,973 --> 04:49:27,483
65 YEARS AGO, AND IT WAS THE 

6379
04:49:32,155 --> 04:49:33,423
BEGINNING OF THE MONOLAYER.

6380
04:49:33,423 --> 04:49:36,726
PLAQUE AND FOCUS ASSAYS HAD ONLY

6381
04:49:36,726 --> 04:49:37,794
BEEN DISCOVERED.

6382
04:49:37,794 --> 04:49:39,128
ANTIBODIES COULD BE GENERATED, 

6383
04:49:39,128 --> 04:49:41,764
BUT ONLY FROM ANIMALS, THERE WAS

6384
04:49:41,764 --> 04:49:47,036
NO BIOCHEMISTRY AND NO MOLECULAR

6385
04:49:47,036 --> 04:49:47,403
SEQUENCING.

6386
04:49:47,403 --> 04:49:49,839
LET'S LOOK AT THE VIRAL SURFACE 

6387
04:49:49,839 --> 04:49:50,173
GLYCOPROTEINS.

6388
04:49:50,173 --> 04:49:53,309
A LOT OF THEM FROM A TO J, SOME 

6389
04:49:53,309 --> 04:49:57,780
ARE EXCLUSIVELY AVIAN HOST 

6390
04:49:57,780 --> 04:50:06,789
RANGE, SOME CAN INFECT MAMMALIAN

6391
04:50:06,789 --> 04:50:09,625
CELLS, AND SOME ARE AVIAN 

6392
04:50:09,625 --> 04:50:11,627
ENDOGENOUS RETROVIRUSES.

6393
04:50:11,627 --> 04:50:13,029
WHAT'S INTERESTING IS THERE IS 

6394
04:50:13,029 --> 04:50:16,165
AN EVOLVING SUBGROUP THAT IS THE

6395
04:50:16,165 --> 04:50:17,567
PREDOMINANT PATHOGEN IN THE 

6396
04:50:17,567 --> 04:50:19,535
POULTRY INDUSTRY IN CHINA, AND 

6397
04:50:19,535 --> 04:50:20,903
IT IS SPREADING VERY RAPIDLY 

6398
04:50:20,903 --> 04:50:23,706
OVER THE GLOBE.

6399
04:50:23,706 --> 04:50:29,812
YOU CAN USE THESE SUBGROUPS TO 

6400
04:50:29,812 --> 04:50:32,315
DESIGN INTERFERENCE ASSAYS, AND 

6401
04:50:32,315 --> 04:50:33,216
INTERFERENCE WITH SUBGROUP 

6402
04:50:33,216 --> 04:50:33,483
SPECIFIC.

6403
04:50:33,483 --> 04:50:36,953
IF A CELL IS INFECTED WITH A 

6404
04:50:36,953 --> 04:50:40,523
SUBGROUP A VIRUS, IT WOULD BE 

6405
04:50:40,523 --> 04:50:42,825
SPECIFICALLY RESISTANT TO A 

6406
04:50:42,825 --> 04:50:48,331
CHALLENGE WITH A VIRUS, AND B 

6407
04:50:48,331 --> 04:50:53,369
MAKES RESISTANT TO B AND SO ON.

6408
04:50:53,369 --> 04:50:55,071
HERE IS AN EXAMPLE.

6409
04:50:55,071 --> 04:50:58,741
IT'S AN AVIAN LEUKOSIS VIRUS 

6410
04:50:58,741 --> 04:51:02,145
THAT INFECTS CELLS WITHOUT ANY 

6411
04:51:02,145 --> 04:51:03,179
MORPHOLOGICAL EVENT.

6412
04:51:03,179 --> 04:51:05,581
YOU WOULD NOT BE ABLE TO TELL 

6413
04:51:05,581 --> 04:51:08,484
THE INFECTED FROM THE 

6414
04:51:08,484 --> 04:51:10,153
NON-INFECTED CULTURES WITHOUT 

6415
04:51:10,153 --> 04:51:13,956
ANY SORT OF BIOCHEMICAL TOOLS, 

6416
04:51:13,956 --> 04:51:16,459
WHICH DIDN'T EXIST AT THAT TIME.

6417
04:51:16,459 --> 04:51:20,963
BUT IF YOU HAVE AN AVIAN 

6418
04:51:20,963 --> 04:51:23,800
LEUKOSIS VIRUS A IN INFECTED 

6419
04:51:23,800 --> 04:51:25,635
CELL LINE AND THEN CHALLENGE 

6420
04:51:25,635 --> 04:51:28,404
WITH A, YOU WILL EXPERIENCE 

6421
04:51:28,404 --> 04:51:28,805
INTERFERENCE.

6422
04:51:28,805 --> 04:51:30,540
THERE'S NO INTERFERENCE WITH 

6423
04:51:30,540 --> 04:51:37,246
SUBGROUP B, C, OR D.

6424
04:51:37,246 --> 04:51:40,249
THAT WAS USED IN, I THINK, 

6425
04:51:40,249 --> 04:51:43,853
IMPORTANT EXPERIMENT THAT SHOWED

6426
04:51:43,853 --> 04:51:46,889
THAT EARLY DNA SYNTHESIS 

6427
04:51:46,889 --> 04:51:48,291
REQUIRED IN RETROVIRAL INFECTION

6428
04:51:48,291 --> 04:51:50,126
IS VIRUS SPECIFIC.

6429
04:51:50,126 --> 04:51:52,228
SO YOU TAKE YOUR CELLS AND 

6430
04:51:52,228 --> 04:51:59,135
INFECT THEM WITH RSV A AS ZERO 

6431
04:51:59,135 --> 04:51:59,335
HOURS.

6432
04:51:59,335 --> 04:52:02,572
AT 24 HOURS YOU INHIBIT THE DNA 

6433
04:52:02,572 --> 04:52:05,074
SYNTHESIS, SO YOU GIVE VIRUS A 

6434
04:52:05,074 --> 04:52:07,310
TIME TO MAKE DNA.

6435
04:52:07,310 --> 04:52:11,581
THEN YOU CHALLENGE AT 96 HOURS 

6436
04:52:11,581 --> 04:52:17,620
WITH RSV C, AND THEN YOU ANALYZE

6437
04:52:17,620 --> 04:52:20,523
THE PROGENY.

6438
04:52:20,523 --> 04:52:24,060
RSV A CAN BE MADE, BUT RSV C 

6439
04:52:24,060 --> 04:52:25,328
CANNOT BE MADE.

6440
04:52:25,328 --> 04:52:27,763
SO THESE TWO VIRUSES CANNOT 

6441
04:52:27,763 --> 04:52:31,534
SHARE THE SAME DNA.

6442
04:52:31,534 --> 04:52:34,003
DNA MADE EARLY IN INFECTION IS 

6443
04:52:34,003 --> 04:52:36,739
VIRUS SPECIFIC.

6444
04:52:36,739 --> 04:52:47,250
SO THAT BRINGS ME TO LESSON 4.

6445
04:52:50,353 --> 04:52:53,522
YOU SHOULD EARLY ON GET TO KNOW 

6446
04:52:53,522 --> 04:52:54,991
PEOPLE IN THE FIELD AND ASK FOR 

6447
04:52:54,991 --> 04:52:55,858
THEIR REAGENTS.

6448
04:52:55,858 --> 04:52:58,628
I DID THAT EARLY ON, AND THEY 

6449
04:52:58,628 --> 04:53:00,496
TURNED OUT TO BE VERY 

6450
04:53:00,496 --> 04:53:00,796
INTERESTING.

6451
04:53:00,796 --> 04:53:03,165
THE PRAGUE STRAIN OF RSV WAS THE

6452
04:53:03,165 --> 04:53:04,400
FIRST, REPRESENTATIVE OF A NEW 

6453
04:53:04,400 --> 04:53:14,610
SUBGROUP C.

6454
04:53:18,748 --> 04:53:22,151
THEN I GOT THE CARR-Z IRBGS LBER

6455
04:53:22,151 --> 04:53:23,853
STRAIN OF RSV, AND 

6456
04:53:23,853 --> 04:53:25,388
NEUROFIBROMATOSIS A NEW SUBGROUP

6457
04:53:25,388 --> 04:53:25,688
D.

6458
04:53:25,688 --> 04:53:29,558
THEN SCHMIDT-RUPPIN SENT ME HIS 

6459
04:53:29,558 --> 04:53:31,160
VIRUS, AND IT WAS ALSO IN 

6460
04:53:31,160 --> 04:53:32,261
SUBGROUP D.

6461
04:53:32,261 --> 04:53:36,432
AND THEN SOMEONE SENT ME THE 

6462
04:53:36,432 --> 04:53:38,034
SARCOMA VIRUS, WHICH WAS VERY, 

6463
04:53:38,034 --> 04:53:40,536
VERY SIMILAR TO OUR SARCOMA 

6464
04:53:40,536 --> 04:53:41,170
VIRUS.

6465
04:53:41,170 --> 04:53:44,907
I SUBMITTED IT WAS IN 

6466
04:53:44,907 --> 04:53:46,008
CZECHOSLOVAKIA AND CALLED IT 

6467
04:53:46,008 --> 04:53:48,311
VIRUS B77 AND ALSO SUBGROUP C.

6468
04:53:48,311 --> 04:53:50,713
SO YOU COULD ENRICH YOUR 

6469
04:53:50,713 --> 04:53:52,949
INVENTORY AND YOUR TOOLS BY 

6470
04:53:52,949 --> 04:53:53,582
COLLECTING SOME VIRUSES FROM 

6471
04:53:53,582 --> 04:53:59,555
OTHER LABS.

6472
04:53:59,555 --> 04:54:01,724
THAT BRINGS ME TO LESSON 5, 

6473
04:54:01,724 --> 04:54:02,858
ATTRACT GREAT POST DOCS.

6474
04:54:02,858 --> 04:54:05,628
I WANT TO BRIEFLY MENTION THREE 

6475
04:54:05,628 --> 04:54:08,364
EXAMP

6476
04:54:08,364 --> 04:54:09,265
EXAMPLES.

6477
04:54:09,265 --> 04:54:11,334
I'LL START AT THE BOTTOM BECAUSE

6478
04:54:11,334 --> 04:54:13,502
IT'S AN HIV MEETING AND YOU ALL 

6479
04:54:13,502 --> 04:54:15,104
WILL KNOW ERIC HUNTER.

6480
04:54:15,104 --> 04:54:18,607
YOU ALSO WILL KNOW ROBIN WEISS 

6481
04:54:18,607 --> 04:54:22,645
BECAUSE HE ALSO WORKED WITH HIV.

6482
04:54:22,645 --> 04:54:27,750
AND THEN THERE'S KUMAO 

6483
04:54:27,750 --> 04:54:29,151
TOYOSHIMA.

6484
04:54:29,151 --> 04:54:30,953
KUMAO WAS ESSENTIALLY THE 

6485
04:54:30,953 --> 04:54:32,955
EMPEROR OF TUMOR VIROLOGY IN 

6486
04:54:32,955 --> 04:54:34,190
JAPAN.

6487
04:54:34,190 --> 04:54:37,626
HE RECEIVED THE HIGHEST 

6488
04:54:37,626 --> 04:54:39,929
DISTINCTION AWARD FROM THE 

6489
04:54:39,929 --> 04:54:40,629
JAPANESE GOVERNMENT.

6490
04:54:40,629 --> 04:54:44,266
HE PASSED AWAY TWO YEARS AGO.

6491
04:54:44,266 --> 04:54:46,335
ROBIN WEISS IS A MEMBER OF THE 

6492
04:54:46,335 --> 04:54:50,306
NATIONAL ACADEMY OF SCIENCES 

6493
04:54:50,306 --> 04:54:50,473
USA.

6494
04:54:50,473 --> 04:54:52,441
HE IS A MEMBER OF THE AMERICAN 

6495
04:54:52,441 --> 04:54:53,709
PHILOSOPHICAL SOCIETY, AND OF 

6496
04:54:53,709 --> 04:54:54,710
COURSE HE IS A MEMBER OF THE 

6497
04:54:54,710 --> 04:54:58,280
ROYAL SOCIETY IN LONDON.

6498
04:54:58,280 --> 04:55:04,754
ERIC HUNTER STILL VERY ACTIVE, 

6499
04:55:04,754 --> 04:55:06,422
AND HE COMES TO A MEETING EVERY 

6500
04:55:06,422 --> 04:55:08,324
YEAR IN JANUARY OUT HERE IN SAN 

6501
04:55:08,324 --> 04:55:09,525
DIEGO, AND WE ALWAYS MEET AND 

6502
04:55:09,525 --> 04:55:14,864
HAVE DINNER TOGETHER.

6503
04:55:14,864 --> 04:55:17,299
LESSON 6 IS TO INTERROGATE 

6504
04:55:17,299 --> 04:55:19,802
NATURE, GO OUT AND COLLECT 

6505
04:55:19,802 --> 04:55:22,471
VIRUSES FROM THE ANIMAL SOURCE.

6506
04:55:22,471 --> 04:55:24,807
SO I WENT INTO THESE CHICKEN 

6507
04:55:24,807 --> 04:55:27,143
SLAUGHTER HOUSES WITH PERMISSION

6508
04:55:27,143 --> 04:55:34,283
OF THE DEPARTMENT OF AGRICULTURE

6509
04:55:34,283 --> 04:55:36,218
AND COLLECTED TUMORS AND 

6510
04:55:36,218 --> 04:55:37,653
ISOLATES VIRUSES.

6511
04:55:37,653 --> 04:55:40,856
I ALSO GOT VIRUSES FROM OTHER 

6512
04:55:40,856 --> 04:55:46,429
COLLECTORS SOME EXAMPLES OF 

6513
04:55:46,429 --> 04:55:49,298
IMPORTANT ONCOGENES THAT EMERGED

6514
04:55:49,298 --> 04:55:53,102
FROM THESE SPONTANEOUS TUMORS, 

6515
04:55:53,102 --> 04:55:57,206
MC29, WHICH IS AN AVIAN LEUKEMIA

6516
04:55:57,206 --> 04:56:00,142
VIRUS ISOLATED OR ACTUALLY 

6517
04:56:00,142 --> 04:56:05,247
COLLECTED BY JOE BEARD AT DUKE 

6518
04:56:05,247 --> 04:56:09,618
UNIVERSITY THAT CONTAINED MYC.

6519
04:56:09,618 --> 04:56:12,188
MYC IS PERHAPS THE EMPEROR OF 

6520
04:56:12,188 --> 04:56:13,689
ONCOGENES, VERY, VERY IMPORTANT 

6521
04:56:13,689 --> 04:56:14,857
IN HUMAN MEDICINE.

6522
04:56:14,857 --> 04:56:18,828
THEN AVIAN SARCOMA VIRUS 17 

6523
04:56:18,828 --> 04:56:20,796
YIELDED JUN, AN IMPORTANT 

6524
04:56:20,796 --> 04:56:25,568
REGULATOR OF TRANSCRIPTION.

6525
04:56:25,568 --> 04:56:29,205
AN NOT COGENE THAT IS DERIVED 

6526
04:56:29,205 --> 04:56:32,341
FROM OTHER AVIAN SARCOMA VIRUS 

6527
04:56:32,341 --> 04:56:34,343
IS QIN.

6528
04:56:34,343 --> 04:56:36,612
IT IS NOW CALLED FOX G.

6529
04:56:36,612 --> 04:56:38,914
IT WAS THE FIRST EXAMPLE OF A 

6530
04:56:38,914 --> 04:56:43,085
FOX PROTEIN ISOLATED FROM A 

6531
04:56:43,085 --> 04:56:43,652
VERTEBRATE.

6532
04:56:43,652 --> 04:56:48,757
AND FINALLY, AVIAN SARCOMA VIRUS

6533
04:56:48,757 --> 04:56:53,095
16 YIELDED PI 3-KINASE.

6534
04:56:53,095 --> 04:56:54,830
SO THAT BRINGS ME TO THE FINAL 

6535
04:56:54,830 --> 04:56:58,534
LESSON, WHICH IS LESSON 7, 

6536
04:56:58,534 --> 04:56:59,068
COLLABORATE, COLLABORATE, 

6537
04:56:59,068 --> 04:57:03,639
COLLABORATE.

6538
04:57:03,639 --> 04:57:05,174
SCIENCE IS DRIVEN BY ADVANCES IN

6539
04:57:05,174 --> 04:57:05,574
TECHNOLOGY.

6540
04:57:05,574 --> 04:57:07,610
YOU WILL NOT BE ABLE TO MASTER 

6541
04:57:07,610 --> 04:57:08,911
ALL NEW TECHNIQUES.

6542
04:57:08,911 --> 04:57:09,512
YOU HAVE TO COLLABORATE WITH 

6543
04:57:09,512 --> 04:57:16,352
EXPERTS.

6544
04:57:16,352 --> 04:57:17,686
I'D LIKE TO CITE BRIEFLY THREE 

6545
04:57:17,686 --> 04:57:17,953
EXAMPLES.

6546
04:57:17,953 --> 04:57:20,222
ONE IS THE DISCOVERY OF THE 

6547
04:57:20,222 --> 04:57:26,095
FIRST ONCOGENE COLLABORATION.

6548
04:57:26,095 --> 04:57:27,029
THE SECOND IS -- OOPS.

6549
04:57:27,029 --> 04:57:37,306
LET ME GO BACK.

6550
04:57:40,242 --> 04:57:43,646
GENERATING THE SRC PROBE BY 

6551
04:57:43,646 --> 04:57:47,049
SUBTRACTIVE HYBRIDIZATION.

6552
04:57:47,049 --> 04:57:51,954
IT'S A CO-STUDY WITH THE BISHOP 

6553
04:57:51,954 --> 04:57:54,156
LAB, AND ONE OF THE MAJOR FORCES

6554
04:57:54,156 --> 04:57:55,791
IN THIS LAB WAS STEPHEN HUGHES.

6555
04:57:55,791 --> 04:57:57,760
SO I WANT TO SAY HI TO STEVE.

6556
04:57:57,760 --> 04:57:58,394
HE IS PROBABLY STILL IN THE 

6557
04:57:58,394 --> 04:58:03,599
AUDIENCE.

6558
04:58:03,599 --> 04:58:07,570
AND THE FINAL IS ON THE CRYO-EM 

6559
04:58:07,570 --> 04:58:10,005
STRUCTURE OF WILD TYPE AND 

6560
04:58:10,005 --> 04:58:16,645
MUTANT PI 3-KINASES.

6561
04:58:16,645 --> 04:58:18,914
SO THE SRC PARADIGM IS REALLY 

6562
04:58:18,914 --> 04:58:21,417
BASED ON THE NON-DEFECTIVENESS 

6563
04:58:21,417 --> 04:58:24,653
OF ROUS SARCOMA VIRUS.

6564
04:58:24,653 --> 04:58:26,689
THIS VIRUS HIGHLY SIMPLIFIED 

6565
04:58:26,689 --> 04:58:28,424
GENOME STRUCTURE CONTAINS ALL 

6566
04:58:28,424 --> 04:58:32,294
THE BIOGENES PLUS SRC.

6567
04:58:32,294 --> 04:58:37,633
AND SRC IS LOST IN REPLICATION, 

6568
04:58:37,633 --> 04:58:43,939
AND THIS DELETION MUTANT 

6569
04:58:43,939 --> 04:58:46,475
ACTUALLY REPLICATES FASTER THAN 

6570
04:58:46,475 --> 04:58:50,446
THE ORIGINAL COMPLETE ROUS 

6571
04:58:50,446 --> 04:58:51,614
SARCOMA VIRUS.

6572
04:58:51,614 --> 04:58:53,115
EVENTUALLY YOU CAN SEPARATE 

6573
04:58:53,115 --> 04:58:55,484
DELETION FROM TRANSFORMING 

6574
04:58:55,484 --> 04:58:55,684
VIRUS.

6575
04:58:55,684 --> 04:58:57,953
IF YOU THEN COMPARE SIZE OF RNA,

6576
04:58:57,953 --> 04:58:59,588
YOU'LL FIND THAT THE RNA OF THE 

6577
04:58:59,588 --> 04:59:01,423
TRANSFORMING VIRUS IS LARGER 

6578
04:59:01,423 --> 04:59:03,926
THAN THE RNA OF THE 

6579
04:59:03,926 --> 04:59:06,562
NON-TRANSFORMING VIRUS.

6580
04:59:06,562 --> 04:59:08,764
THE CONCLUSION, SUGGESTED 

6581
04:59:08,764 --> 04:59:09,798
CONCLUSION IS THAT THE 

6582
04:59:09,798 --> 04:59:13,936
DIFFERENCE IS IN THE ONCOGENIC 

6583
04:59:13,936 --> 04:59:16,138
TRANSFORMATION IN THE ONCOGENE.

6584
04:59:16,138 --> 04:59:19,842
THAT BRINGS ME TO THE 

6585
04:59:19,842 --> 04:59:20,542
SUBTRAC

6586
04:59:20,542 --> 04:59:23,912
SUBTRACTIVE HYBRIDIZATION THAT 

6587
04:59:23,912 --> 04:59:26,715
BISHOP PERFORMED WITH THIS SAME 

6588
04:59:26,715 --> 04:59:27,449
SET OF VIRUSES.

6589
04:59:27,449 --> 04:59:30,452
YOU HAVE THE RNA OF A 

6590
04:59:30,452 --> 04:59:33,055
NON-DEFECTIVE ROUS SARCOMA 

6591
04:59:33,055 --> 04:59:33,255
VIRUS.

6592
04:59:33,255 --> 04:59:36,125
YOU TRANSCRIBE IT WITH REVERSE 

6593
04:59:36,125 --> 04:59:37,860
TRANSCRIPTORS INTO SINGLE STRAND

6594
04:59:37,860 --> 04:59:40,429
DNA, USUALLY IN SMALL PIECES, 

6595
04:59:40,429 --> 04:59:45,567
AND HYBRIDIZE THAT RNA IN 

6596
04:59:45,567 --> 04:59:46,468
TRANSFORMATION-DEFECTIVE ROUS 

6597
04:59:46,468 --> 04:59:47,503
SARCOMA VIRUS GENOME.

6598
04:59:47,503 --> 04:59:50,739
THAT CAPTURES ALL THE VIRAL 

6599
04:59:50,739 --> 04:59:54,943
SEQUENCES, AND YOU ARE LEFT WITH

6600
04:59:54,943 --> 04:59:56,912
SINGLE STRAND OF DNA THAT IS 

6601
04:59:56,912 --> 04:59:59,348
SPECIFIC FOR THE ONCOGENE.

6602
04:59:59,348 --> 05:00:02,751
THAT'S THE SRC ROPE.

6603
05:00:02,751 --> 05:00:04,853
AND THAT LED TO THIS REALLY 

6604
05:00:04,853 --> 05:00:06,655
IMPORTANT CONCLUSION THAT ALL 

6605
05:00:06,655 --> 05:00:09,425
RETROVIRAL ONCOGENES ARE 

6606
05:00:09,425 --> 05:00:10,826
CELLULAR GENES ARE HITCHHIKERS 

6607
05:00:10,826 --> 05:00:15,898
ON THE VIRAL GENOME.

6608
05:00:15,898 --> 05:00:17,533
THAT BRINGS ME TO THE 

6609
05:00:17,533 --> 05:00:24,973
COLLABORATION OF PI3-KINASE.

6610
05:00:24,973 --> 05:00:29,978
PI3-KINASE IS A DIMERIC LIPID 

6611
05:00:29,978 --> 05:00:30,212
KINASE.

6612
05:00:30,212 --> 05:00:33,115
THE ISOFORM IMPORTANT FOR HUMAN 

6613
05:00:33,115 --> 05:00:36,652
CANCER IS THE A ISOFORM.

6614
05:00:36,652 --> 05:00:38,587
IT CONSISTS OF CATALYTIC 

6615
05:00:38,587 --> 05:00:40,089
SUBUNITS AND REGULATORY 

6616
05:00:40,089 --> 05:00:40,356
SUBUNITS.

6617
05:00:40,356 --> 05:00:41,824
THERE HAVE BEEN MANY, MANY 

6618
05:00:41,824 --> 05:00:44,660
ATTEMPTS TO GET THE CRYO-M 

6619
05:00:44,660 --> 05:00:46,028
STRUCTURE OF THAT PROTEIN, AND 

6620
05:00:46,028 --> 05:00:47,529
ALL OF THEM FAILED.

6621
05:00:47,529 --> 05:00:50,299
WHEN WE STARTED, WE WERE TOLD TO

6622
05:00:50,299 --> 05:00:52,468
FORGET ABOUT IT, THAT WE WERE 

6623
05:00:52,468 --> 05:00:56,972
NEW BYES IN THIS FIELD, WE WOULD

6624
05:00:56,972 --> 05:00:57,940
NEVER SUCCEED BECAUSE MANY OF 

6625
05:00:57,940 --> 05:01:00,008
THE REAL EXPERTS HAD NOT BEEN 

6626
05:01:00,008 --> 05:01:06,348
ABLE TO DO IT.

6627
05:01:06,348 --> 05:01:07,983
BUT THAT ADVICE IGNORED THE FACT

6628
05:01:07,983 --> 05:01:11,653
THAT IN THE U.S. ACCESS TO 

6629
05:01:11,653 --> 05:01:15,557
CRYO-EM IS VERY LIMITED AND VERY

6630
05:01:15,557 --> 05:01:17,760
EXPENSIVE, AND WE COULDN'T GET 

6631
05:01:17,760 --> 05:01:22,564
ENOUGH TIME IN CRYO-EM SCRIPTS 

6632
05:01:22,564 --> 05:01:24,166
TO DO THAT WORK.

6633
05:01:24,166 --> 05:01:25,534
SO EVENTUALLY I GOT TOGETHER 

6634
05:01:25,534 --> 05:01:29,905
WITH A FRIEND IN SHANGHAI, MING 

6635
05:01:29,905 --> 05:01:31,874
HOO WON, AND WE DECIDED TO DO IT

6636
05:01:31,874 --> 05:01:33,375
HERE, THREE SCOPES, UNLIMITED 

6637
05:01:33,375 --> 05:01:36,011
TIME, WE COULD TRIAL AND ERROR 

6638
05:01:36,011 --> 05:01:39,281
APPROACH IT, USE TRIAL AND ERROR

6639
05:01:39,281 --> 05:01:42,317
APPROACHES TO SEE WHAT WORKS.

6640
05:01:42,317 --> 05:01:44,520
THERE WERE LOTS OF DIFFICULTIES.

6641
05:01:44,520 --> 05:01:50,092
FIRST OF ALL, NCI WOULDN'T 

6642
05:01:50,092 --> 05:01:51,527
APPROVE, AND WE HAD TO BEG AND 

6643
05:01:51,527 --> 05:01:55,063
BEG AND BEG, AND EVENTUALLY OUR 

6644
05:01:55,063 --> 05:01:58,233
REQUEST WENT UP ALL THE WAY TO 

6645
05:01:58,233 --> 05:02:02,070
THE EMBASSY IN BEIJING, THE U.S.

6646
05:02:02,070 --> 05:02:04,173
EMBASSY IN BEIJING, AND THEY 

6647
05:02:04,173 --> 05:02:06,275
SAID, OKAY, YOU CAN GO AHEAD.

6648
05:02:06,275 --> 05:02:09,044
THEN WE GOT APPROVAL, AND WE 

6649
05:02:09,044 --> 05:02:12,581
STARTED WORK WITH THE SHANGHAI 

6650
05:02:12,581 --> 05:02:13,215
TEAM.

6651
05:02:13,215 --> 05:02:15,350
MONTHS OF MONTHS OF MONTHS OF NO

6652
05:02:15,350 --> 05:02:17,519
SUCCESS, AND FINALLY WE HIT THE 

6653
05:02:17,519 --> 05:02:19,755
RIGHT FREEZING CONDITIONS, THE 

6654
05:02:19,755 --> 05:02:21,223
RIGHT BUFFER CONDITIONS, AND WE 

6655
05:02:21,223 --> 05:02:24,059
WERE SUCCESSFUL.

6656
05:02:24,059 --> 05:02:27,129
I JUST WANT TO SHOW YOU THE 

6657
05:02:27,129 --> 05:02:34,470
ELECTRON DENSITY MAP OF P

6658
05:02:34,470 --> 05:02:34,770
PI3-KINASE.

6659
05:02:34,770 --> 05:02:36,104
WE WERE ABLE TO IDENTIFY EACH OF

6660
05:02:36,104 --> 05:02:38,373
THE DOMAINS OF THESE PROTEINS, 

6661
05:02:38,373 --> 05:02:40,542
AND NOT ONLY THE WILD TYPE, BUT 

6662
05:02:40,542 --> 05:02:41,810
ALSO THE CANCER ASSOCIATED 

6663
05:02:41,810 --> 05:02:44,313
MUTANTS, AND WE USED THAT 

6664
05:02:44,313 --> 05:02:48,083
INFORMATION TO GENERATE 

6665
05:02:48,083 --> 05:02:49,485
MUTANT-SPECIFIC LIGANDS THAT 

6666
05:02:49,485 --> 05:02:50,452
COULD BE DEVELOPED INTO 

6667
05:02:50,452 --> 05:02:52,821
INHIBITORS.

6668
05:02:52,821 --> 05:02:55,657
THAT'S REALLY ALL I WANT TO SAY,

6669
05:02:55,657 --> 05:02:58,327
AND I WANT TO END BY SAYING 

6670
05:02:58,327 --> 05:02:59,361
HAPPY RETIREMENT, ALAN.

6671
05:02:59,361 --> 05:03:09,538
THANK YOU.

6672
05:03:31,760 --> 05:03:32,728
>> THANK YOU, PETER.

6673
05:03:32,728 --> 05:03:34,363
THANK YOU FOR A WONDERFUL TALK.

6674
05:03:34,363 --> 05:03:35,297
EXCELLENT ADVICE TO ALL YOU 

6675
05:03:35,297 --> 05:03:41,236
YOUNG KIDS DOWN THERE.

6676
05:03:41,236 --> 05:03:43,138
I'M WISHING YOU REALLY HAPPY, 

6677
05:03:43,138 --> 05:03:49,578
PRODUCTIVE YEARS AHEAD.

6678
05:03:49,578 --> 05:03:51,980
AND WHAT'S NEXT ON THE PROGRAM 

6679
05:03:51,980 --> 05:04:02,324
IS CLOSING REMARKS.

6680
05:04:09,298 --> 05:04:10,666
I HAVE A FEW THINGS I WANT TO 

6681
05:04:10,666 --> 05:04:10,832
SAY.

6682
05:04:10,832 --> 05:04:12,901
FIRST OF ALL, I COULD NEVER 

6683
05:04:12,901 --> 05:04:14,670
BELIEVE THAT I WOULD LIVE TO SEE

6684
05:04:14,670 --> 05:04:15,571
AN EVENT LIKE THIS.

6685
05:04:15,571 --> 05:04:18,006
I REALLY APPRECIATE ALL THE KIND

6686
05:04:18,006 --> 05:04:20,175
THINGS THAT EVERYONE HAS SAID 

6687
05:04:20,175 --> 05:04:28,650
AND THE FACT THAT AND THE EFFORT

6688
05:04:28,650 --> 05:04:30,986
THAT WAS MADE TO PRODUCE THIS 

6689
05:04:30,986 --> 05:04:31,453
EVENT.

6690
05:04:31,453 --> 05:04:33,021
THANK YOU TO YOU ALL FOR COMING.

6691
05:04:33,021 --> 05:04:34,823
I SPECIFICALLY WANT TO THANK THE

6692
05:04:34,823 --> 05:04:36,725
PEOPLE WHO MADE THIS EVENT 

6693
05:04:36,725 --> 05:04:36,992
POSSIBLE.

6694
05:04:36,992 --> 05:04:38,427
THAT'S ERIC WHO SUGGESTED IT, 

6695
05:04:38,427 --> 05:04:40,495
WHICH AGAIN WAS A COMPLETE 

6696
05:04:40,495 --> 05:04:42,731
SURPR

6697
05:04:42,731 --> 05:04:44,499
SURPRISE, AND SPECIFICALLY ANNA 

6698
05:04:44,499 --> 05:04:47,869
NORRIS AND ALSO TERRY BURDETT.

6699
05:04:47,869 --> 05:04:50,305
NO DOUBT OTHER PEOPLE BEHIND THE

6700
05:04:50,305 --> 05:04:51,306
SCENES WHOSE NAMES I MAY NOT 

6701
05:04:51,306 --> 05:04:59,815
EVEN KNOW.

6702
05:04:59,815 --> 05:05:01,083
SECONDLY, I WANT TO GIVE CREDIT 

6703
05:05:01,083 --> 05:05:03,852
TO A NUMBER OF OTHER PEOPLE FOR 

6704
05:05:03,852 --> 05:05:07,522
DIFFERENT CONTRIBUTIONS TO MY 

6705
05:05:07,522 --> 05:05:09,858
TRAJECTORY AND TO THE FACT THAT 

6706
05:05:09,858 --> 05:05:13,962
I'M HERE TODAY.

6707
05:05:13,962 --> 05:05:17,099
I HAVE BEEN VERY FORTUNATE IN MY

6708
05:05:17,099 --> 05:05:21,103
CHOICE OF BOSSES, AND I WANTED 

6709
05:05:21,103 --> 05:05:23,972
TO MENTION, FIRST OF ALL, BOB 

6710
05:05:23,972 --> 05:05:29,277
BASSEN, WHO INTRODUCED ME TO THE

6711
05:05:29,277 --> 05:05:31,279
MURINE LEUKEMIA VIRUS AND WAS 

6712
05:05:31,279 --> 05:05:33,782
ALWAYS GENEROUS AND SUPPORTIVE.

6713
05:05:33,782 --> 05:05:35,984
I'M SURE FRANCOIS WOULD AGREE.

6714
05:05:35,984 --> 05:05:38,820
SHE MAY HAVE GONE TO BED BY NOW,

6715
05:05:38,820 --> 05:05:40,222
BUT ANYWAY, I'M SURE SHE WOULD 

6716
05:05:40,222 --> 05:05:42,524
AGREE.

6717
05:05:42,524 --> 05:05:46,161
WE WERE ALL VERY CLOSE TO BOB.

6718
05:05:46,161 --> 05:05:50,065
HE AND, AS I THINK FRANCOIS 

6719
05:05:50,065 --> 05:05:52,100
MENTIONED, HE DIED A YEAR AGO 

6720
05:05:52,100 --> 05:05:53,368
QUITE UNEXPECTEDLY.

6721
05:05:53,368 --> 05:06:03,512
SO I'M SORRY HE'S NOT HERE.

6722
05:06:03,512 --> 05:06:07,449
AFTER BOB, I WAS HIRED IN 

6723
05:06:07,449 --> 05:06:11,019
FREDERICK BY REGULAN, WHO AGAIN 

6724
05:06:11,019 --> 05:06:12,721
WAS HELPFUL AND SUPPORTIVE AND 

6725
05:06:12,721 --> 05:06:14,456
HELPED ESTABLISH GOOD RESEARCH 

6726
05:06:14,456 --> 05:06:15,691
HERE IN FREDERICK.

6727
05:06:15,691 --> 05:06:26,201
THEN GEORGE VANDRUD, WHO PUT 

6728
05:06:26,568 --> 05:06:30,906
FREDERICK ON THE MAP AS A FIRST 

6729
05:06:30,906 --> 05:06:32,574
CLASS SITE FOR RESEARCH.

6730
05:06:32,574 --> 05:06:34,943
GEORGE HAD A VERY BROAD VISION 

6731
05:06:34,943 --> 05:06:45,487
ABOUT CANCER RESEARCH AND HOW TO

6732
05:06:55,130 --> 05:06:55,464
FOSTER RESEARCH.

6733
05:06:55,464 --> 05:06:57,165
ONE OF THE THINGS GEORGE DID WAS

6734
05:06:57,165 --> 05:06:58,533
FOSTER THE CREATION OF THE DRP, 

6735
05:06:58,533 --> 05:07:00,769
AND THAT WAS REALLY THE CREATION

6736
05:07:00,769 --> 05:07:09,177
OF JOHN COFFIN, SO JOHN SET UP 

6737
05:07:09,177 --> 05:07:09,778
THE DRP.

6738
05:07:09,778 --> 05:07:10,879
AND STEVE AND I WERE ALREADY 

6739
05:07:10,879 --> 05:07:12,681
HERE WHEN THE DRP WAS FORMED.

6740
05:07:12,681 --> 05:07:14,549
EVERYONE ELSE IN THE DRP WAS 

6741
05:07:14,549 --> 05:07:16,952
REALLY RECRUITED BY JOHN OR 

6742
05:07:16,952 --> 05:07:17,753
SUBSEQUENTLY, BUT MAINLY 

6743
05:07:17,753 --> 05:07:23,925
RECRUITED BY JOHN.

6744
05:07:23,925 --> 05:07:27,262
THEN EVENTUALLY JOHN HANDED THE 

6745
05:07:27,262 --> 05:07:32,167
BATON TO STEVE, AND EVENTUALLY 

6746
05:07:32,167 --> 05:07:33,969
AFTER THAT, STEVE HANDED THE 

6747
05:07:33,969 --> 05:07:35,637
BATON TO ERIC, AND ALL THREE OF 

6748
05:07:35,637 --> 05:07:38,240
THEM, ONE AFTER THE OTHER, HAVE 

6749
05:07:38,240 --> 05:07:42,077
BEEN AGAIN IMMENSELY SUPPORTIVE 

6750
05:07:42,077 --> 05:07:46,014
AND HELPFUL AND REALLY FOSTERED 

6751
05:07:46,014 --> 05:07:50,485
GOOD RESEARCH AND GOOD FREE 

6752
05:07:50,485 --> 05:07:54,389
THINKING.

6753
05:07:54,389 --> 05:08:03,431
SO I WANT TO THANK ALL OF THEM.

6754
05:08:03,431 --> 05:08:05,167
I ALSO JUST WANT TO MENTION A 

6755
05:08:05,167 --> 05:08:07,602
COUPLE OF PEOPLE WHO HAVE MADE 

6756
05:08:07,602 --> 05:08:08,303
ENORMOUS CONTRIBUTIONS IN THE 

6757
05:08:08,303 --> 05:08:09,604
RESEARCH IN THE LAB, SO I HAD 

6758
05:08:09,604 --> 05:08:12,240
TWO TECHNICIANS, ONE WHO JOINED 

6759
05:08:12,240 --> 05:08:16,178
IN 1989 AND ONE WHO JOINED IN 

6760
05:08:16,178 --> 05:08:20,649
1990, AND THE ONE WHO JOINED IN 

6761
05:08:20,649 --> 05:08:22,818
1989 RETIRED A FEW YEARS AGO, 

6762
05:08:22,818 --> 05:08:25,620
JANE MIRRO, AND THE ONE WHO 

6763
05:08:25,620 --> 05:08:27,255
JOINED IN 1990 JUST RETIRED 

6764
05:08:27,255 --> 05:08:30,025
ABOUT A YEAR AGO, AND SHE IS 

6765
05:08:30,025 --> 05:08:39,434
HERE TODAY, THAT'S DIMITRIA 

6766
05:08:39,434 --> 05:08:43,238
HARVIN, KNOWN TO US AS MEECH.

6767
05:08:43,238 --> 05:08:44,606
THESE TWO KEPT THE LAB RUNNING 

6768
05:08:44,606 --> 05:08:46,074
AND KEPT THE EXPERIMENTS GOING, 

6769
05:08:46,074 --> 05:08:47,375
AND REALLY BEAR A LOT OF 

6770
05:08:47,375 --> 05:08:48,410
RESPONSIBILITY FOR THE 

6771
05:08:48,410 --> 05:08:50,312
ACCOMPLISHMENTS OF THE LAB.

6772
05:08:50,312 --> 05:08:54,716
THEN I MUST MAKE SPECIAL MENTION

6773
05:08:54,716 --> 05:08:59,654
OF SID DABEH, SO HE JOINED THE 

6774
05:08:59,654 --> 05:09:08,830
LAB IN 2001, AND HE CAME WITH 

6775
05:09:08,830 --> 05:09:10,298
TREMENDOUS EXPERTISE ABOUT 

6776
05:09:10,298 --> 05:09:12,267
PROTEIN STRUCTURE, PROTEIN 

6777
05:09:12,267 --> 05:09:16,304
FUNCTION, PROTEIN PURIFICATION, 

6778
05:09:16,304 --> 05:09:20,242
AND HE HAS REALLY, I THINK, 

6779
05:09:20,242 --> 05:09:22,677
PLAYED A CRITICAL ROLE IN ALL 

6780
05:09:22,677 --> 05:09:28,283
THE WORK IN THE LAB SINCE HE 

6781
05:09:28,283 --> 05:09:29,117
JOINED IN 2001.

6782
05:09:29,117 --> 05:09:30,085
SO MUCH OF THE WORK IN THE LAB 

6783
05:09:30,085 --> 05:09:32,520
WAS WORKING DIRECTLY WITH 

6784
05:09:32,520 --> 05:09:33,855
PROTEINS PURIFIED FROM BACTERIA.

6785
05:09:33,855 --> 05:09:44,266
THAT WAS VERY IMPORTANT.

6786
05:09:47,269 --> 05:09:48,169
SHIFT IN DIRECTION OF OUR 

6787
05:09:48,169 --> 05:09:50,338
RESEARCH, BUT NOT ALL OF IT.

6788
05:09:50,338 --> 05:09:52,274
SOME OF IT WAS GOOD OLD 

6789
05:09:52,274 --> 05:09:53,742
FASHIONED VIROLOGY PERFORMED IN 

6790
05:09:53,742 --> 05:09:55,543
THE MILLION CELLS, BUT SID ALSO 

6791
05:09:55,543 --> 05:09:58,213
PLAYED A CRITICAL ROLE, JUST AS 

6792
05:09:58,213 --> 05:10:03,018
ANOTHER CRITICAL THINKER AND 

6793
05:10:03,018 --> 05:10:04,386
TRYING TO UNDERSTAND WHAT'S THE 

6794
05:10:04,386 --> 05:10:05,186
RIGHT EXPERIMENT?

6795
05:10:05,186 --> 05:10:06,454
WHAT DOES THIS RESULT MEAN?

6796
05:10:06,454 --> 05:10:14,596
WHAT DO WE DO NEXT?

6797
05:10:14,596 --> 05:10:16,197
SO I WANT EVERYONE TO UNDERSTAND

6798
05:10:16,197 --> 05:10:19,134
HOW IMPORTANT SID'S WORK HAS 

6799
05:10:19,134 --> 05:10:20,602
BEEN, SID HAS BEEN IN THE 

6800
05:10:20,602 --> 05:10:21,670
ACCOMPLISHMENTS OF THE LAB FOR 

6801
05:10:21,670 --> 05:10:23,171
THE LAST 20-PLUS YEARS.

6802
05:10:23,171 --> 05:10:25,907
BEYOND THAT, I WANTED TO -- I 

6803
05:10:25,907 --> 05:10:32,681
HAVE BEEN, NEEDLESS TO SAY, 

6804
05:10:32,681 --> 05:10:38,153
RETROSPECTING, INTROSPECTING, 

6805
05:10:38,153 --> 05:10:39,554
HOW DID I GET HERE?

6806
05:10:39,554 --> 05:10:41,556
WHAT LED ME TO THIS JUNCTURE OF 

6807
05:10:41,556 --> 05:10:43,391
HAVING SPENT ALL THESE YEARS 

6808
05:10:43,391 --> 05:10:44,826
STUDYING WHAT WE NOW CALL 

6809
05:10:44,826 --> 05:10:55,070
RETROVIRUSES?

6810
05:10:58,173 --> 05:11:00,108
SO I WILL TAKE A COUPLE OF 

6811
05:11:00,108 --> 05:11:01,876
MINUTES TO RETRACE MY STEPS FOR 

6812
05:11:01,876 --> 05:11:02,043
YOU.

6813
05:11:02,043 --> 05:11:04,112
I'VE THOUGHT ABOUT THEM, AND 

6814
05:11:04,112 --> 05:11:05,914
POSSIBLY WHAT I'M GOING TO SAY, 

6815
05:11:05,914 --> 05:11:09,084
ALTHOUGH VERY SIMPLISTIC, SIMPLE

6816
05:11:09,084 --> 05:11:12,153
MINDED, NAIVE, MAY NEVERTHELESS 

6817
05:11:12,153 --> 05:11:13,855
RESONATE WITH MANY OF YOU.

6818
05:11:13,855 --> 05:11:15,890
SO I WENT TO HIGH SCHOOL IN THE 

6819
05:11:15,890 --> 05:11:25,767
1950s, AND FOR CONTEXT, THE 

6820
05:11:25,767 --> 05:11:26,634
WATSON-CRITZ STRUCTURE, WHICH IS

6821
05:11:26,634 --> 05:11:28,870
A FOUNDATION OF WHAT WE DO EVER 

6822
05:11:28,870 --> 05:11:30,105
SINCE, WAS PUBLISHED IN 1953, AS

6823
05:11:30,105 --> 05:11:34,909
YOU KNOW.

6824
05:11:34,909 --> 05:11:37,746
AND I THINK I PROBABLY READ 

6825
05:11:37,746 --> 05:11:39,481
ABOUT THE DNA STRUCTURE IN 

6826
05:11:39,481 --> 05:11:41,750
SCIENTIFIC AMERICAN, AND I TOLD 

6827
05:11:41,750 --> 05:11:46,621
MY PARENTS THAT I WANTED TO BE A

6828
05:11:46,621 --> 05:11:46,921
BIOCHEMIST.

6829
05:11:46,921 --> 05:11:48,123
THEY SAID, YOU DON'T WANT TO BE 

6830
05:11:48,123 --> 05:11:49,891
A BIOCHEMIST.

6831
05:11:49,891 --> 05:11:52,761
THEY KNEW SOMEONE WHO WAS A 

6832
05:11:52,761 --> 05:11:53,061
BIOCHEMIST.

6833
05:11:53,061 --> 05:11:57,699
AND HE SAID, HE TOLD THEM -- 

6834
05:11:57,699 --> 05:12:00,435
>> DON'T LET ALAN BE A 

6835
05:12:00,435 --> 05:12:00,735
BIOCHEMIST.

6836
05:12:00,735 --> 05:12:03,438
>> DON'T LET ALAN BE A 

6837
05:12:03,438 --> 05:12:05,340
BIOCHEMIST.

6838
05:12:05,340 --> 05:12:07,342
BIOCHEMISTRY, HE SAID -- THIS IS

6839
05:12:07,342 --> 05:12:10,612
A QUOTE -- IS A LIFE OF 

6840
05:12:10,612 --> 05:12:14,749
PROGRESSIVE POVERTY.

6841
05:12:14,749 --> 05:12:17,419
ALAN SHOULD BE A DOCTOR INSTEAD.

6842
05:12:17,419 --> 05:12:17,786
(LAUGHTER) 

6843
05:12:17,786 --> 05:12:21,122
BUT NO, I WANTED TO BE A 

6844
05:12:21,122 --> 05:12:23,658
BIOCHEMIST COME HELL OR HIGH 

6845
05:12:23,658 --> 05:12:24,092
WATER.

6846
05:12:24,092 --> 05:12:28,530
I WANTED TO BE A BIOCHEMIST.

6847
05:12:28,530 --> 05:12:33,501
SO THAT'S WHAT I WANTED TO DO.

6848
05:12:33,501 --> 05:12:34,469
WHY IS THAT?

6849
05:12:34,469 --> 05:12:36,271
WHY DID I WANT TO BE A 

6850
05:12:36,271 --> 05:12:36,571
BIOCHEMIST?

6851
05:12:36,571 --> 05:12:38,873
IT'S BECAUSE, I THINK -- WELL, I

6852
05:12:38,873 --> 05:12:40,475
TOOK, LIKE EVERYBODY ELSE, I 

6853
05:12:40,475 --> 05:12:43,178
TOOK CHEMISTRY IN HIGH SCHOOL.

6854
05:12:43,178 --> 05:12:43,778
I LIKED CHEMISTRY IN HIGH 

6855
05:12:43,778 --> 05:12:48,583
SCHOOL.

6856
05:12:48,583 --> 05:12:50,919
YOU FIND OUT THE ATOMS, THE 

6857
05:12:50,919 --> 05:12:52,353
MOLECULES, THEY FOLLOW VERY 

6858
05:12:52,353 --> 05:12:53,054
SIMPLE RULES.

6859
05:12:53,054 --> 05:12:55,490
THAT'S WHAT CHEMISTRY IS.

6860
05:12:55,490 --> 05:12:57,692
WHAT ARE THE ATOMS AND MOLECULES

6861
05:12:57,692 --> 05:12:58,827
GOING TO DO BECAUSE OF COURSE 

6862
05:12:58,827 --> 05:13:00,195
THEY FOLLOW THE RULES.

6863
05:13:00,195 --> 05:13:01,529
THAT'S WHAT CHEMISTRY IS.

6864
05:13:01,529 --> 05:13:02,831
AT THE SAME TIME, IF YOU STUDY 

6865
05:13:02,831 --> 05:13:10,271
THE MOLECULES IN LIVING THINGS, 

6866
05:13:10,271 --> 05:13:12,040
THEY SEEM TO ACT WITH A PURPOSE.

6867
05:13:12,040 --> 05:13:13,908
THEY SEEM TO HAVE FUNCTION.

6868
05:13:13,908 --> 05:13:15,977
YOU KNOW MOLECULES, IT'S LIKE 

6869
05:13:15,977 --> 05:13:18,379
THEY KNOW WHAT THEY'RE DOING.

6870
05:13:18,379 --> 05:13:19,914
BUT SINCE YOU KNOW A LITTLE BIT 

6871
05:13:19,914 --> 05:13:21,349
ABOUT CHEMISTRY, YOU KNOW THEY 

6872
05:13:21,349 --> 05:13:23,618
DON'T KNOW WHAT THEY'RE DOING.

6873
05:13:23,618 --> 05:13:25,820
THEY'RE JUST FOLLOWING RULES.

6874
05:13:25,820 --> 05:13:30,091
I THINK THIS BASIC PARADOX IS 

6875
05:13:30,091 --> 05:13:33,261
WHAT ATTRACTED ME.

6876
05:13:33,261 --> 05:13:37,332
HOW IS THIS POSSIBLE THAT THESE 

6877
05:13:37,332 --> 05:13:38,433
MOLECULES, WHICH ARE ONLY 

6878
05:13:38,433 --> 05:13:39,767
FOLLOWING RULES, SEEM TO KNOW 

6879
05:13:39,767 --> 05:13:43,004
WHAT THEY'RE DOING AND ACT WITH 

6880
05:13:43,004 --> 05:13:51,079
PURPOSE?

6881
05:13:51,079 --> 05:13:53,848
SO NOT TAKING MY PARENTS' GOOD 

6882
05:13:53,848 --> 05:13:55,150
ADVICE, I WENT TO COLLEGE.

6883
05:13:55,150 --> 05:13:56,684
I STUDIED BIOLOGY IN COLLEGE, 

6884
05:13:56,684 --> 05:13:58,553
AND THEN WHEN I WENT TO GRADUATE

6885
05:13:58,553 --> 05:14:00,054
SCHOOL, I CHOSE BERKELEY, AND 

6886
05:14:00,054 --> 05:14:04,359
THE REASON I CHOSE BERKELEY WAS 

6887
05:14:04,359 --> 05:14:05,894
I DID NOT KNOW EXACTLY WHAT I 

6888
05:14:05,894 --> 05:14:09,230
WANTED TO STUDY, AND I FIGURED 

6889
05:14:09,230 --> 05:14:11,266
BERKELEY WOULD HAVE GOOD 

6890
05:14:11,266 --> 05:14:12,233
POSSIBILITIES IN JUST ABOUT 

6891
05:14:12,233 --> 05:14:13,902
ANYTHING I WANTED TO STUDY.

6892
05:14:13,902 --> 05:14:16,271
I WAS KEEPING MY OPTIONS OPEN BY

6893
05:14:16,271 --> 05:14:18,139
GOING TO BERKELEY.

6894
05:14:18,139 --> 05:14:20,475
THINK THAT WAS EXACTLY RIGHT.

6895
05:14:20,475 --> 05:14:24,646
I STARTED OFF IN BERKELEY IN A 

6896
05:14:24,646 --> 05:14:29,450
VERY FAMOUS, RESPECTED LAB, 

6897
05:14:29,450 --> 05:14:35,557
STUDYING SEA URCHIN EMBRYOLOGY, 

6898
05:14:35,557 --> 05:14:36,658
WHICH WAS VERY INTERESTING.

6899
05:14:36,658 --> 05:14:38,459
BUT THERE WAS A GUY UP THE HILL,

6900
05:14:38,459 --> 05:14:40,862
HARRY RUBIN, STUDYING THESE 

6901
05:14:40,862 --> 05:14:41,763
AMAZING VIRUSES.

6902
05:14:41,763 --> 05:14:43,898
YOU JUST PUT THE VIRUSES ON THE 

6903
05:14:43,898 --> 05:14:45,800
CELLS, A COUPLE DAYS LATER, 

6904
05:14:45,800 --> 05:14:47,702
THEY'RE CANCER CELLS.

6905
05:14:47,702 --> 05:14:50,271
ISN'T THAT FANTASTIC?

6906
05:14:50,271 --> 05:14:53,141
SO I WAS JUST -- IT WAS JUST UP 

6907
05:14:53,141 --> 05:14:58,146
THE HILL, AND I SWITCHED TO 

6908
05:14:58,146 --> 05:15:00,114
RUBIN'S LAB.

6909
05:15:00,114 --> 05:15:02,717
HERE WE ARE, ALL THESE DECADES 

6910
05:15:02,717 --> 05:15:02,917
LATER.

6911
05:15:02,917 --> 05:15:05,787
STILL TALKING ABOUT WHAT WE NOW 

6912
05:15:05,787 --> 05:15:11,793
CALL RETROVIRUSES.

6913
05:15:11,793 --> 05:15:14,596
I THINK I BECAME A VIROLOGIST 

6914
05:15:14,596 --> 05:15:17,131
BECAUSE VIROLOGISTS JUST 

6915
05:15:17,131 --> 05:15:19,167
EPITOMIZE THIS PARADOX, THIS 

6916
05:15:19,167 --> 05:15:19,834
CONTRADICTION.

6917
05:15:19,834 --> 05:15:21,035
ON THE ONE HAPPENED, THEY'RE 

6918
05:15:21,035 --> 05:15:22,170
JUST A FEW MOLECULES.

6919
05:15:22,170 --> 05:15:23,471
THEY'RE JUST SITTING THERE.

6920
05:15:23,471 --> 05:15:25,373
ON THE OTHER HAND, AS WE ALL 

6921
05:15:25,373 --> 05:15:28,109
KNOW, AS WE'VE SPENT OUR LIVES 

6922
05:15:28,109 --> 05:15:29,344
STUDYING, THOSE MOLECULES SURE 

6923
05:15:29,344 --> 05:15:30,712
SEEM TO ACT WITH A PURPOSE.

6924
05:15:30,712 --> 05:15:32,947
SO IT WAS THE FASCINATION WITH 

6925
05:15:32,947 --> 05:15:40,321
THIS PARADOX THAT HAS HAS DRIVEN

6926
05:15:40,321 --> 05:15:41,389
ME ALL THESE YEARS.

6927
05:15:41,389 --> 05:15:43,157
I STILL FIND IT FASCINATING, AND

6928
05:15:43,157 --> 05:15:44,993
I THINK THAT'S WHAT I WANTED TO 

6929
05:15:44,993 --> 05:15:45,526
SAY.

6930
05:15:45,526 --> 05:15:47,862
THANK YOU ALL FOR COMING AND ALL

6931
05:15:47,862 --> 05:15:51,332
THE KIND WORDS.

6932
05:15:51,332 --> 05:15:53,901
I WANT TO ESPECIALLY THANK ERIC.

6933
05:15:53,901 --> 05:15:56,137
I DID NOT EXPECT AN EVENT LIKE 

6934
05:15:56,137 --> 05:16:00,642
THIS, AND IT'S REALLY BEEN MORE 

6935
05:16:00,642 --> 05:16:02,076
THAN I CAN -- I APPRECIATE IT 

6936
05:16:02,076 --> 05:16:05,546
MORE THAN I CAN EXPRESS.

6937
05:16:05,546 --> 05:16:06,214
THANK YOU AGAIN.

6938
05:16:06,214 --> 05:16:07,248
NOW ERIC HAS SOME CLOSING, 

6939
05:16:07,248 --> 05:16:17,525
CLOSING REMARKS.

6940
05:16:22,997 --> 05:16:24,465
>> YOU'RE ALL PROBABLY
SAYING, 

6941
05:16:24,465 --> 05:16:27,168
WAIT A SECOND, ERIC'S NOT ON THE

6942
05:16:27,168 --> 05:16:27,402
PROGRAM.

6943
05:16:27,402 --> 05:16:28,903
HE ALREADY TALKED TODAY, AND I 

6944
05:16:28,903 --> 05:16:30,838
REALLY WANT TO GO TO STEINHART'S

6945
05:16:30,838 --> 05:16:32,006
AND GET A BEER.

6946
05:16:32,006 --> 05:16:33,341
I WILL BE VERY BRIEF.

6947
05:16:33,341 --> 05:16:36,244
I JUST WANTED TO JOIN ALAN IN 

6948
05:16:36,244 --> 05:16:37,745
THANKING ALL OF YOU FOR COMING 

6949
05:16:37,745 --> 05:16:38,946
FROM NEAR AND FROM FAR.

6950
05:16:38,946 --> 05:16:40,782
THOSE OF YOU WHO JOINED REMOTELY

6951
05:16:40,782 --> 05:16:42,283
AND MIGHT STILL BE ON THE CALL, 

6952
05:16:42,283 --> 05:16:43,051
A LOT OF GOOD DISCUSSION IN THE 

6953
05:16:43,051 --> 05:16:45,820
AUDIENCE.

6954
05:16:45,820 --> 05:16:48,990
MOST OF ALL, I'D LIKE TO THANK 

6955
05:16:48,990 --> 05:16:49,290
ALAN.

6956
05:16:49,290 --> 05:16:50,925
I THINK WE GOT A FLAVOR TODAY OF

6957
05:16:50,925 --> 05:16:52,960
THE RESPECT, ADMIRATION, AND 

6958
05:16:52,960 --> 05:16:53,594
AFFECTION THAT THE ENTIRE 

6959
05:16:53,594 --> 05:16:55,396
COMMUNITY HAS FOR ALAN.

6960
05:16:55,396 --> 05:16:57,999
NOW I'D REALLY LIKE TO THANK YOU

6961
05:16:57,999 --> 05:17:00,034
FOR BEING MY COLLEAGUE ALL THESE

6962
05:17:00,034 --> 05:17:01,202
MANY YEARS.

6963
05:17:01,202 --> 05:17:02,937
I'VE REALLY CHERISHED IT.

6964
05:17:02,937 --> 05:17:04,439
I WISH YOU THE BEST OF LUCK IN 

6965
05:17:04,439 --> 05:17:06,174
YOUR FUTURE, AND I HOPE FOR MANY

6966
05:17:06,174 --> 05:17:10,078
MORE INTERACTIONS IN THE YEARS 

6967
05:17:10,078 --> 05:17:10,278
AHEAD.

6968
05:17:10,278 --> 05:17:12,347
ONE LAST THING OTHER THAN 

6969
05:17:12,347 --> 05:17:13,514
MENTIONING AGAIN THERE IS A 

6970
05:17:13,514 --> 05:17:15,950
GATHERING AT STEINHART'S.

6971
05:17:15,950 --> 05:17:17,719
CORINNE MENTIONED THE RETROVIRUS

6972
05:17:17,719 --> 05:17:20,121
ASSEMBLY MEETING IN PRAGUE, THE 

6973
05:17:20,121 --> 05:17:22,156
DATES ARE SEPTEMBER 16-19 OF 

6974
05:17:22,156 --> 05:17:23,424
NEXT YEAR.

6975
05:17:23,424 --> 05:17:25,727
YOU CAN GOOGLE PRAGUE RETROVIRUS

6976
05:17:25,727 --> 05:17:27,295
ASSEMBLY MEETING, AND IT WILL 

6977
05:17:27,295 --> 05:17:28,930
COME RIGHT UP, AND YOU CAN 

6978
05:17:28,930 --> 05:17:29,997
REGISTER THERE.

6979
05:17:29,997 --> 05:17:32,433
I HOPE TO SEE MANY OF YOU THERE.

6980
05:17:32,433 --> 05:17:42,944
AND THANK YOU AGAIN VERY MUCH.