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>> GOOD MORNING, EVERYONE.

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I'M A COLLEGIATE DAVID McDONALD 

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AT THE BASIC SCIENCE PROGRAM AND

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THANK YOU ALL FOR A WONDERFUL 

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PRESENTATION YESTERDAY AND IT'S 

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GOOD TO SEE NEW SCIENCE AND 

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WE'RE IN THE SECOND DAY AND WE 

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HAVE THREE MORE CENTERS 

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PRESENTING TODAY.

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I JUST WANTED BEFORE WE GO THERE

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SOME FEW HOUSEKEEPING 

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ANNOUNCEMENT.

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YOU SAW A SLIDE A MOMENT EARLIER

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THE NEXT MEETING PLEASE MARK 

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YOUR CALENDAR.

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LOOKING FORWARD TO IT NEXT YEAR 

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AND PEOPLE ARE PRESENTING 

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ABSTRACTS, PLEASE COME UP BEFORE

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LUNCH AND GIVE YOUR PRESENTATION

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SO WE COULD BE READY AND COME 

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BACK AFTER LUNCH IN THE POSTER 

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SESSION.

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WITH THAT LET ME INTRODUCE THE 

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FIRST CENTER PRESENTER TODAY AND

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WE'LL HAVE A PRESENT BY ALICE 

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TELESNITSKY.

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>> WE'RE THE CENTER FOR 

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STRUCTURAL BIOLOGY OF HIV RNA.

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AS THE NAME IMPLIES THE PROJECTS

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THAT WE'RE WORKING ON HAVE DO 

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WITH INTERACTIONS OF MOLECULAR 

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COMPLEXES AND PROCESSES IN THE 

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HIV REPLICATION CYCLE THAT 

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INVOLVE RNA.

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WE HAVE A FABULOUS GROUP OF 

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INVESTIGATORS AND COLLABORATIVE 

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AWARD WINNERS AND 

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INTERGENERATIONAL AND 

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INTER-CENTER INTERACTIONS AND 

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I'LL SKIP THOSE AND HIGHLIGHT 

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ONE OF OUR ACTIVITIES WHICH IS 

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WE HAVE AN UNDER GRADUATE 

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RESEARCH PROGRAM -- AH, WE CALL 

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IT THE HIV OPPORTUNITY IN RNA 

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FOR UNDERGRADUATE SCHOLARS WE 

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CALL ROAR.

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VERY CUTE NAME, HUH.

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WE CAN SEE THE SECOND YEAR OF 

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THE PROGRAM AND TRAINEES ARE 

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EMBEDDED.

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THIS IS MY FAVORITE TRAINING LAB

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AND JEREMY LUBAN AND I WERE 

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TRAINEES AND HE IS HOSTING ONE 

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OF OUR ROAR STUDENTS THIS YEAR 

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AND THEY'RE ALREADY EMBEDDED IN 

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THEIR LABS BUT BECAUSE LIKE 

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OTHER CENTERS WE'RE REMOTE AND 

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IT WAS JUST NICE THEY COULD COME

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TO THIS MEETING AND INTERACT 

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WITH YOU SO YOU'LL HAVE SEEN 

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SOME OF THEM AT THE POSTERS 

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YESTERDAY AND TODAY.

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I WANTED TO CUT STRAIGHT TO THE 

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SCIENCE BECAUSE WE HAVE EXCITING

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THINGS TO TELL YOU ABOUT TODAY 

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AND FIRST UP IS PAUL BENASH.

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-- PAUL BIENIASZ.

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I'LL TELL YOU A COLLABORATIVE 

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LAB ON THE ZINC FINGER COMPLEX 

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BASED ON A FOUNDATIONAL RESULT 

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IF YOU TAKE HIV-1 AND TAKE A 

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SMALL SEGMENT OF ITS GENOME AND 

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BY SYNONYMOUS SUBSTITUTION 

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ELEVATE THE CPG CONTENT TO 

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APPROXIMATELY THE LEVEL YOU'D 

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SEE IN RANDOM RNA SEQUENCE 

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IMPOSES A LETHAL DEFECT IMPOSED 

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BY THE PRESENCE OF THE ZAP 

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PROTEIN NOT ALLOWING THE VIRUS 

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TO REPLICATE LIKE WILD TYPE.

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AND ZINC PROTEIN ROSE IN AN 

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ANCESTRAL AQUAPOD AFTER THE 

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LINEAGE OF TRANSFERASES WHICH 

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LED TO THE PURGING OF CPG WILL 

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NUCLEOTIDES FROM THE 

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TRANSCRIPTOMES OF THAT LINEAGE 

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CREATING AN OPPORTUNITY FOR 

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SELF/NON-SELF DISCRIMINATION BY 

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RECOGNITION OF CPGs CARRIED BY 

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THE VIRUSES OF WILL WILL INSECTS

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AND ARTHROPODS WHICH IS NOT 

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UNDER GO THIS TRANSFERASE.

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BECAUSE THIS EVENT HAPPENED 

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HUNDREDS OF MILLIONS OF TIMES 

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PER DAY FOR HUNDREDS OF MILLIONS

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OF YEARS MAMMALS HAVE BEEN 

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BOMBARDED BY CPG RICH VIRUSES 

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BUT ONLY THE ONES THAT MANAGED 

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TO PURGE CPG FROM THEIR GENOME 

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HAVE BEEN SUCCESSFUL IN 

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COLONIZING MAMMALIAN OF HOSTS.

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WE THINK IT'S LARGELY DO YOU TO 

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THE PROTEIN OF ZAP.

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IT OCCURS IN TWO NATURALLY 

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OCCURRING ISOFORMS.

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THE BUSINESS END MUCH THE 

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PROTEIN IS THIS AMINO TERMINAL 

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BINDING DOMAIN THAT JANET SMITH 

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LAB'S SOLVED THE STRUCTURE OF 

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THE PROTEIN IN COMPLEX WITH AN 

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RNA TARGET WHICH EXPLAINS IN 

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ATOMIC DETAIL HOW THE PROTEIN 

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SPECIFICALLY INTERACTS WITH CPG 

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DINUCLEOTIDES AND THE SINGLE 

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EVENT IS NOT ENOUGH FOR THE 

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ANTIVIRAL ACTIVITY AND LEARNED 

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FROM GROUPS IMPLICATING A NUMBER

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OF PROTEINS BUT MOST 

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COMPELLINGLY AND THE PROTEINS 

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I'LL TALK ABOUT TODAY, TRIM 25 

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AND K9  KHNYN IMPORTANT AND 

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ESSENTIAL FOR THE ACTIVITY OF 

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ZAP.

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OUR OWN AND I'LL SHOW DATA WHERE

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WE DO A SIMPLE CYCLE EXPERIMENT 

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AND MEASURE WITHIN A DAY OR TWO 

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HOW MUCH VIRUS COMES ON YOU IN 

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CELLS KNOCKED OUT FOR ONE OF 

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THESE PROTEINS THAT MAY OR MAY 

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NOT BE INVOLVED.

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SO HERE'S THE BASELINE 

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PHENOTYPE.

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A VIRUS THAT HAS A SEGMENT OF CP

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G ENRICHED CONSEQUENCE.

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IT HAS A 50 FOLD SEQUENCE OVER 

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REPLICATION AND COMPLETELY GONE 

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WHEN YOU KNOCK OUT ZAP.

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IT'S DOWN BUT NOT OUT.

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IF YOU KNOCK OUT THIS CO-FACTOR 

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TRIM 25 IS UNAFFECTED IF YOU 

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KNOCK OUT THE NUCLEASE KHNYN AND

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KNOCK OUT A RELATED FAMILY 

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MEMBER LITTLE AFFECT BUT IF YOU 

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KNOCK OUT BOTH THEN THE 

145
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PHENOTYPE GOES AWAY.

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IT LOOKS LIKE THE PROTEINS ACT 

147
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REDUNDANTLY AND WE CAN 

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RECONSTITUTE THE DOUBLE 

149
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KNOCKOUTS WITH ONE OR EITHER OF 

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THE PROTEINS.

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SO ARMED WITH THAT KNOWLEDGE WE 

152
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DEVELOPED A RECONSTITUTION 

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ASSAY.

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A SIMPLE TRANSFECTION ASSAY 

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WHERE WE GENERATED A CELL LINE 

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KNOCKED OUT FOR ALL THE PLAYERS 

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WILL INTERESTED AND TRANSFECT 

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PROVIRAL DNA WITH OR WITHOUT ONE

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OF THE PROTEINS IN WILD TYPE OR 

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MUTANT FORM AND MEASURE 

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INFECTIOUS VIRUS.

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IN THIS PARTICULAR ITERATION WE 

163
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HAVE WILD TYPE HIV-1 WITH THE CV

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ENRICHED SEGMENT AND YOU CAN SEE

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AU  YOU NEED ALL THREE TO GET IT

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A COUPLE ORDERS OF MAGNITUDE OF 

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INFECTIOUS VIRUS RELEASE.

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INDIVIDUAL PROTEINS ON THEIR OWN

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DO NOTHING IN THIS 

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RECONSTITUTION SYSTEM AND KHNYN 

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HAS SOME ACTIVITY AND CURIOUSLY 

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SOMEWHAT OF A LOSS OF 

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SPECIFICITY AND HAVE WEAK 

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ACTIVITY AGAINST THE WILD TYPE 

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VIRUS.

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SO I'M GOING TO SHOW YOU A 

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NUMBER OF EXPERIMENTS USING THAT

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RECONSTITUTION SYSTEM.

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HERE FIRST WE'RE TAKING THE ZAP 

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PROTEIN AND DOING SIMPLE 

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TRUNCATION EXPERIMENTS AND YOU 

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CAN SHOW AS STEVE GOS' LAB DID 

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THE AMINO FERM NAL FRAGMENT THE 

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RNA BINDING DOMAIN IS SUFFICIENT

185
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FOR ACTIVITY.

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YOU GET INHIBITION OF THE VIRUS 

187
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AND SOME INHIBITION OF THE WILD 

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TYPE VIRUS.

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YOU NEED LONGER FORMS AND 

190
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ISOFORMS TO GET THIS 

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SPECIFICITY.

192
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YOU CAN SEE A HIGH DEGREE OF 

193
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SPECIFICITY FOR CG ENRICHED 

194
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VIRUS WITH THE FULL LENGTH 

195
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PROTEIN COMPARED AMINO TERMINAL 

196
00:09:03,867 --> 00:09:04,501
FRAGMENT.

197
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THE OTHER ESSENTIAL COMPONENT IS

198
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AS I SAID A NUCLEASE AND USE THE

199
00:09:11,875 --> 00:09:21,151
KHNYN FOR THESE EXPERIMENTS.

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S THIS IS THE PUTATIVE DOMAIN 

201
00:09:31,227 --> 00:09:33,463
AND YOU HAVE NUCLEIC ACID DOMAIN

202
00:09:33,463 --> 00:09:35,732
COME TO THAT POINT IN A MINUTE 

203
00:09:35,732 --> 00:09:37,934
AND YOU HAVE C TERMINAL DOMAIN 

204
00:09:37,934 --> 00:09:39,202
OF UNKNOWN FUNCTION.

205
00:09:39,202 --> 00:09:42,005
YOU NEED ALL THESE DOMAINS TO 

206
00:09:42,005 --> 00:09:44,974
GET FULL ACTIVITY OF THE KHNYN 

207
00:09:44,974 --> 00:09:45,642
PROTEIN.

208
00:09:45,642 --> 00:09:48,678
ANY OF THESE TRUNCATION OR POINT

209
00:09:48,678 --> 00:09:51,481
MUTATIONS ABOLISH FOR SEVERELY 

210
00:09:51,481 --> 00:09:54,217
ABLATE THE ACTIVITY.

211
00:09:54,217 --> 00:09:56,052
THAT C TERMINAL DOMAIN SEEMS TO 

212
00:09:56,052 --> 00:10:01,057
BE AN IMPORTANT PART OF 

213
00:10:01,057 --> 00:10:02,092
ASSEMBLING THE COMPLEX AND THIS 

214
00:10:02,092 --> 00:10:04,894
WAS SOLVED IN JANET SMITH'S LAP 

215
00:10:04,894 --> 00:10:08,164
WITH THE RBD DOMAIN AND THAT 

216
00:10:08,164 --> 00:10:12,435
SMALL KHNYN C TERMINAL DOMAIN.

217
00:10:12,435 --> 00:10:14,404
YOU CAN SEE THE INTERFACE IF YOU

218
00:10:14,404 --> 00:10:17,740
LOOK IN DETAIL WE CAN IDENTIFY 

219
00:10:17,740 --> 00:10:22,145
BASICALLY THREE POINTS OF 

220
00:10:22,145 --> 00:10:26,316
CONTACT BETWEEN KHNYN AND ZAP 

221
00:10:26,316 --> 00:10:28,618
AND THIS IS NOT THE ONLY ONE 

222
00:10:28,618 --> 00:10:30,854
IT'S AN IMPORTANT ONE AND IF YOU

223
00:10:30,854 --> 00:10:33,256
MUTATE THIS PARTICULAR AMINO 

224
00:10:33,256 --> 00:10:36,960
ACID AT INTERFACE 1 YOU CAN 

225
00:10:36,960 --> 00:10:37,660
COMPROMISE DRAMATICALLY THE 

226
00:10:37,660 --> 00:10:38,228
ABILITY.

227
00:10:38,228 --> 00:10:41,197
SORRY, THIS IS A ZAP MUTATION 

228
00:10:41,197 --> 00:10:42,298
YOU CAN COMPROMISE THE ANTIVIRAL

229
00:10:42,298 --> 00:10:49,005
ACTIVITY.

230
00:10:49,005 --> 00:10:52,509
IMPORTANTLY THE KHNYN CTD 

231
00:10:52,509 --> 00:10:54,110
INTERACTION OCCURS AT A 

232
00:10:54,110 --> 00:10:55,912
DIFFERENT PLACE THAN THE RNA 

233
00:10:55,912 --> 00:10:58,314
BINDING AND IT'S FULLY 

234
00:10:58,314 --> 00:11:02,318
COMPATIBLE WITH ASSEMBLY OF RNA,

235
00:11:02,318 --> 00:11:03,586
ZAP AND KHNYN TOGETHER.

236
00:11:03,586 --> 00:11:06,456
SO I'VE CALLED KHNYN A PUTATIVE 

237
00:11:06,456 --> 00:11:07,390
NUCLEASE.

238
00:11:07,390 --> 00:11:09,859
IS IT ACTUALLY A NUCLEASE?

239
00:11:09,859 --> 00:11:12,695
THE ANSWER IS YES.

240
00:11:12,695 --> 00:11:15,465
HERE'S THE FULL LENGTH PROTEIN 

241
00:11:15,465 --> 00:11:19,469
WITH APPROXIMATELY 100 

242
00:11:19,469 --> 00:11:22,238
NUCLEOTIDE WITH CG DINUCLEOTIDES

243
00:11:22,238 --> 00:11:24,274
OR DOESN'T HAVE THAT HAVE BEEN 

244
00:11:24,274 --> 00:11:26,442
REVERSED TO GCs.

245
00:11:26,442 --> 00:11:27,710
YOU CAN SEE THE NUCLEASE DOESN'T

246
00:11:27,710 --> 00:11:29,412
REALLY CARE ABOUT THE PRESENCE 

247
00:11:29,412 --> 00:11:32,282
OF CGs IN THE RNA SUBSTRATE.

248
00:11:32,282 --> 00:11:33,850
IT WILL DIGEST COMPLETELY REMOVE

249
00:11:33,850 --> 00:11:37,854
THE RNA IN ABOUT 20 MINUTES.

250
00:11:37,854 --> 00:11:42,025
NOW, IF YOU EXPRESS JUST THE 

251
00:11:42,025 --> 00:11:44,160
WILL NIN DOMAIN THAT ALSO IS AN 

252
00:11:44,160 --> 00:11:45,828
ACTIVE NUCLEASE AND ALSO DOESN'T

253
00:11:45,828 --> 00:11:48,398
CARE ABOUT THE NUCLEOTIDE 

254
00:11:48,398 --> 00:11:58,508
CONTENT OF THE TARGET RNA.

255
00:11:58,508 --> 00:12:00,176
AND HAS LIMITING AMOUNT OF 

256
00:12:00,176 --> 00:12:02,645
NUCLEASE AND YOU CAN SEE THE 

257
00:12:02,645 --> 00:12:05,181
PRODUCTS BUT IF YOU ASSEMBLE THE

258
00:12:05,181 --> 00:12:07,584
DUPLEX IT'S COMPLETELY 

259
00:12:07,584 --> 00:12:08,918
RESISTANT.

260
00:12:08,918 --> 00:12:10,853
SINGLE STRAND RESISTANT 

261
00:12:10,853 --> 00:12:11,120
NUCLEASE.

262
00:12:11,120 --> 00:12:14,290
MORE DETAILS HOW THIS COMPLEX 

263
00:12:14,290 --> 00:12:18,294
MIGHT INTERACT WITH RNA.

264
00:12:18,294 --> 00:12:20,396
WHERE IN FACT WOULD KHNYN BIND 

265
00:12:20,396 --> 00:12:26,302
ON THE FIRST EXPERIMENT THIS IS 

266
00:12:26,302 --> 00:12:29,839
A POLARIZATION EXPERIMENT AND 

267
00:12:29,839 --> 00:12:33,776
DOESN'T DIGEST THE RNA BUT CAN 

268
00:12:33,776 --> 00:12:41,684
AFFECT THE FLOUROPHORS AND THE 

269
00:12:41,684 --> 00:12:48,958
POLARIZATION IS INCREASED TO I A

270
00:12:48,958 --> 00:12:50,293
GREATER DEGREE SHOWING THE THREE

271
00:12:50,293 --> 00:12:52,128
PRIME POSITION.

272
00:12:52,128 --> 00:12:57,066
IF YOU DO A LIMITING QUANTITY OF

273
00:12:57,066 --> 00:13:00,236
NUCLEASE DIGESTION ASSAY AND ASK

274
00:13:00,236 --> 00:13:03,473
WHY THE FIRST CUT YOU CAN SEE 

275
00:13:03,473 --> 00:13:06,743
THE CUT IS INITIATING THREE 

276
00:13:06,743 --> 00:13:10,280
PRIME TO THE CG BINDING SITE.

277
00:13:10,280 --> 00:13:16,519
THAT SUGGESTS A MODEL FOR THE 

278
00:13:16,519 --> 00:13:18,388
CORE ESSENTIAL MINIMAL PART OF 

279
00:13:18,388 --> 00:13:22,292
THE ANTIVIRAL BINDING DOMAIN 

280
00:13:22,292 --> 00:13:27,363
THAT RECRUITS THE KHNYN CTD AND 

281
00:13:27,363 --> 00:13:30,300
POSITIONS IT TO CLEAVE THE RNA A

282
00:13:30,300 --> 00:13:32,835
FEW NUCLEOTIDES AND THREE PRIME 

283
00:13:32,835 --> 00:13:34,804
TO THE DINUCLEOTIDE.

284
00:13:34,804 --> 00:13:37,740
WE CAN TEST WHETHER THIS IS 

285
00:13:37,740 --> 00:13:40,710
FUNCTIONALLY SUFFICIENT TO 

286
00:13:40,710 --> 00:13:42,211
RECONSTITUTE IN AN IN VIVO 

287
00:13:42,211 --> 00:13:42,412
ASSAY.

288
00:13:42,412 --> 00:13:44,547
WHAT WE'VE DONE IS DONE AWAY 

289
00:13:44,547 --> 00:13:46,382
WITH ALL THE PROTEIN DOMAINS 

290
00:13:46,382 --> 00:13:48,384
THAT WOULD OTHERWISE BE 

291
00:13:48,384 --> 00:13:50,620
ESSENTIAL AND SIMPLY FUSE THE 

292
00:13:50,620 --> 00:13:53,890
ZAP RNA BINDING DOMAIN TO THE IT

293
00:13:53,890 --> 00:13:55,925
NYN DOMAIN OF KHNYN.

294
00:13:55,925 --> 00:13:59,295
WHAT YOU GET WHEN YOU DO THAT 

295
00:13:59,295 --> 00:14:02,332
RATHER REDUCTION EXPERIMENT IS A

296
00:14:02,332 --> 00:14:06,536
VERY POTENT ANTIVIRAL PROTEIN 

297
00:14:06,536 --> 00:14:12,642
AND ENRICH BY TWO ORDERS OF 

298
00:14:12,642 --> 00:14:12,909
MAGNITUDE.

299
00:14:12,909 --> 00:14:15,511
YOU DO LOSE SPECIFICITY SO THE 

300
00:14:15,511 --> 00:14:17,113
WILD TYPE VIRUS IS INHIBITED 

301
00:14:17,113 --> 00:14:17,947
SIGNIFICANTLY BUT IMPORTANTLY 

302
00:14:17,947 --> 00:14:21,517
THE ACTIVITY IS DEPENDENT ON THE

303
00:14:21,517 --> 00:14:25,221
NUCLEASE ACTIVITY OF THE KHNYN 

304
00:14:25,221 --> 00:14:25,655
PART.

305
00:14:25,655 --> 00:14:28,024
WHAT DO THE OTHER COMPONENTS OF 

306
00:14:28,024 --> 00:14:29,292
THE COMPLEX DO?

307
00:14:29,292 --> 00:14:31,594
IS LET'S LOOK AT TRIM 25.

308
00:14:31,594 --> 00:14:33,529
BECAUSE THAT'S OF SOME INTEREST.

309
00:14:33,529 --> 00:14:35,631
SO IN THIS EXPERIMENT WE HAVE 

310
00:14:35,631 --> 00:14:37,433
DONE THE RECONSTITUTION ASSAY ON

311
00:14:37,433 --> 00:14:41,104
WILD TYPE OR CG ENRICHED HIV.

312
00:14:41,104 --> 00:14:43,506
THIS TIME WE'RE ADDING FIXED 

313
00:14:43,506 --> 00:14:46,075
AMOUNT OF KHNYN AND ADDING IN 

314
00:14:46,075 --> 00:14:51,080
TRIM 25 AND SEE THE WILD TYPE 

315
00:14:51,080 --> 00:14:52,849
STIMULATES ANTIVIRAL ACTIVITY 

316
00:14:52,849 --> 00:14:53,850
NICELY.

317
00:14:53,850 --> 00:14:56,552
THE TERMINUS IS IMPORTANT AND 

318
00:14:56,552 --> 00:15:01,991
THE AMINO TERMINUS IS IMPORTANT 

319
00:15:01,991 --> 00:15:12,535
FOR THIS ACTIVITY THIS COMPLEX 

320
00:15:22,712 --> 00:15:29,519
WAS SOLVED WITH THE UBIQUITIN 

321
00:15:29,519 --> 00:15:37,994
AND MAKE MUTATION WERE SHOWN TO 

322
00:15:37,994 --> 00:15:40,696
ABOLISH THE UBIQUITIN TO THE 

323
00:15:40,696 --> 00:15:41,697
ABILITY TO STIMULATE THE 

324
00:15:41,697 --> 00:15:43,299
ACTIVITY AND IF WE MAKE 

325
00:15:43,299 --> 00:15:46,302
MUTATIONS THAT ABOLISH THE 

326
00:15:46,302 --> 00:15:48,337
DIMERIZATION THEN YOU SEVERELY 

327
00:15:48,337 --> 00:15:49,739
COMPROMISE THE ABILITY OF TRIM 

328
00:15:49,739 --> 00:15:52,675
25 TO STIMULATE THE ACTIVITY.

329
00:15:52,675 --> 00:15:56,145
CLEARLY THE ABILITY OF TRIM 25 

330
00:15:56,145 --> 00:15:59,115
TO WILL CREATE HIGH ORDER MULT 

331
00:15:59,115 --> 00:16:00,716
IMERS IS IMPORTANT.

332
00:16:00,716 --> 00:16:02,418
AS FAR AS THE SPRY DOMAIN IS 

333
00:16:02,418 --> 00:16:04,220
CONCERN, IT BECOMES COMPLETELY 

334
00:16:04,220 --> 00:16:06,989
DISPENSABLE IF YOU SIMPLY GET 

335
00:16:06,989 --> 00:16:13,529
RID OF IT AND FUSE INSTEAD THE 

336
00:16:13,529 --> 00:16:17,166
ZAP RNA BINDING DOMAIN AND 

337
00:16:17,166 --> 00:16:20,236
REDUCE IT TO A TWO IT COMPONENT 

338
00:16:20,236 --> 00:16:22,572
SYSTEM AND GET RID OF THE DOMAIN

339
00:16:22,572 --> 00:16:25,775
WHOSE FUNCTION IS RECRUITMENT BY

340
00:16:25,775 --> 00:16:27,510
PUTTING THE RNA BINDING DOMAIN 

341
00:16:27,510 --> 00:16:30,680
YOU CAN SEE THAT ACTIVITY HERE.

342
00:16:30,680 --> 00:16:33,916
AGAIN IT'S VERY POTENT AND NOT 

343
00:16:33,916 --> 00:16:36,853
AS SPECIFIC AS THE FULL COMPLEX 

344
00:16:36,853 --> 00:16:38,287
BUT HAS A HIGH LEVEL OF 

345
00:16:38,287 --> 00:16:41,023
ACTIVITY.

346
00:16:41,023 --> 00:16:45,528
LET ME SUMMARIZE WHAT WE KNOW IN

347
00:16:45,528 --> 00:16:49,398
TERMS OF THE BITS THAT COMPRISE 

348
00:16:49,398 --> 00:16:51,534
THE STRUCTURE OF THIS PURPORTED 

349
00:16:51,534 --> 00:16:52,668
ANTIVIRAL COMPLEX.

350
00:16:52,668 --> 00:16:55,571
I'VE SHOWN THE RBD DOMAIN OF ZAP

351
00:16:55,571 --> 00:16:57,507
INTERACTS WITH KHNYN IN 

352
00:16:57,507 --> 00:17:00,176
PARTICULAR THE C TERMINAL 

353
00:17:00,176 --> 00:17:02,311
DOMAIN.

354
00:17:02,311 --> 00:17:06,716
THAT FORMS THIS CORE COMPLEX OF 

355
00:17:06,716 --> 00:17:09,519
KHNYN AND THE RNA TARGET.

356
00:17:09,519 --> 00:17:11,354
HOWEVER, THAT IS SUFFICIENT FOR 

357
00:17:11,354 --> 00:17:14,290
ACTIVITY BUT NOT COMPLETELY 

358
00:17:14,290 --> 00:17:16,559
SUFFICIENT FOR SPECIFICITY.

359
00:17:16,559 --> 00:17:19,262
THERE ARE ACCESSORY ACTIVITIES 

360
00:17:19,262 --> 00:17:21,163
THAT WE THINK REALLY CONTRIBUTE 

361
00:17:21,163 --> 00:17:22,265
TO THAT SPECIFICITY.

362
00:17:22,265 --> 00:17:26,269
THE FIRST OF THESE IS THE C 

363
00:17:26,269 --> 00:17:28,004
TERMINAL PORTION OF ZAP.

364
00:17:28,004 --> 00:17:31,207
SO TO SOLVE A STRUCTURE OF PART 

365
00:17:31,207 --> 00:17:36,045
OF THAT IS WWE DOMAIN BINDS TO 

366
00:17:36,045 --> 00:17:39,982
POLY ADP RIVOS AND THAT COULD BE

367
00:17:39,982 --> 00:17:47,256
AN IMPORTANT PART OF DRIVING THE

368
00:17:47,256 --> 00:17:49,959
MULTIMERIZATION AND JANET SOLVED

369
00:17:49,959 --> 00:17:52,528
THE KH DOMAIN INVOLVED IN 

370
00:17:52,528 --> 00:17:53,863
NUCLEIC ACID BINDING THAT IS 

371
00:17:53,863 --> 00:17:56,465
CLEARLY NOT THE CASE HERE.

372
00:17:56,465 --> 00:18:01,337
THE KH DOMAIN IS ACIDIC AND DOES

373
00:18:01,337 --> 00:18:03,172
NOT BIND TO RNA IT FORMS PART OF

374
00:18:03,172 --> 00:18:05,675
THE INTERACTION WHICH I HAVEN'T 

375
00:18:05,675 --> 00:18:12,315
SHOWN YOU WITH TRIM 25 AND WITH 

376
00:18:12,315 --> 00:18:12,481
ZAP.

377
00:18:12,481 --> 00:18:14,016
TRIM 25 ITSELF WOULD BE A DIMER 

378
00:18:14,016 --> 00:18:22,792
BUT CAN ASSEMBLE TO HIGHER ORDER

379
00:18:22,792 --> 00:18:26,095
MULTIMERS IMPORTANT FOR THE 

380
00:18:26,095 --> 00:18:27,663
ENHANCING ACTIVITY OF ZAP.

381
00:18:27,663 --> 00:18:30,299
SO AND THAT'S ESSENTIALLY WHAT 

382
00:18:30,299 --> 00:18:33,502
WE KNOW ABOUT THE PROTEIN 

383
00:18:33,502 --> 00:18:34,470
COMPONENTS OF THIS ANTIVIRAL 

384
00:18:34,470 --> 00:18:34,770
COMPLEX.

385
00:18:34,770 --> 00:18:36,572
BEFORE I COME TO A MODEL OF HOW 

386
00:18:36,572 --> 00:18:38,007
WE ALL THINK THIS FITS TOGETHER 

387
00:18:38,007 --> 00:18:40,209
LET ME TALK JUST FOR A FEW 

388
00:18:40,209 --> 00:18:41,877
MINUTES ABOUT THE NATURE OF THE 

389
00:18:41,877 --> 00:18:45,281
TARGET RNA BECAUSE I THINK 

390
00:18:45,281 --> 00:18:46,082
THAT'S QUITE IMPORTANT FOR 

391
00:18:46,082 --> 00:18:47,216
CONSIDERATION.

392
00:18:47,216 --> 00:18:50,753
WHAT IS AN OPTIMAL TARGET OF 

393
00:18:50,753 --> 00:18:50,920
ZAP?

394
00:18:50,920 --> 00:18:58,294
THE DEFINED STRUCTURE IS A 

395
00:18:58,294 --> 00:18:59,729
MONOVALENT OF MOLECULE AND USING

396
00:18:59,729 --> 00:19:01,464
HIV AS A MODEL WE FIGURED OUT 

397
00:19:01,464 --> 00:19:05,601
THE RULES FOR WHAT CONSTITUTES 

398
00:19:05,601 --> 00:19:10,539
AN OPTIMAL ZAP TARGET AND THE 

399
00:19:10,539 --> 00:19:12,441
NUMBER OF AND WHAT'S IMPORTANT 

400
00:19:12,441 --> 00:19:15,778
IS THE SPACING BETWEEN THE CPGs 

401
00:19:15,778 --> 00:19:17,980
ABOUT 12 TO 30 NUCLEOTIDES AND 

402
00:19:17,980 --> 00:19:19,982
THE SINGLE STRANDED CHARACTER IN

403
00:19:19,982 --> 00:19:23,386
WHICH THOSE NUCLEOTIDES ARE 

404
00:19:23,386 --> 00:19:23,653
EMBEDDED.

405
00:19:23,653 --> 00:19:24,787
WE KNOW THESE RULES WELL ENOUGH 

406
00:19:24,787 --> 00:19:28,924
TO DESIGN VIRUSES AND WE'VE DONE

407
00:19:28,924 --> 00:19:33,429
IT WITH THIS VIRUS THAT BECOMES 

408
00:19:33,429 --> 00:19:34,530
ZAP SENSITIVE AND KNOCKOUT CELLS

409
00:19:34,530 --> 00:19:40,002
AND THE VIRUS WE INCREASED THE 

410
00:19:40,002 --> 00:19:42,605
SINGLE STRANDED CHARACTER AND IT

411
00:19:42,605 --> 00:19:45,041
BECOMES ZAP SENSITIVE IN CELL 

412
00:19:45,041 --> 00:19:46,142
CULTURE IN MICE.

413
00:19:46,142 --> 00:19:49,612
THIS IS A ZAP KNOCKOUT MICE AND 

414
00:19:49,612 --> 00:19:51,380
IT WILL REPLICATE WHEN ZAP IS 

415
00:19:51,380 --> 00:19:51,981
KNOCKED OUT AND KILLED THOSE 

416
00:19:51,981 --> 00:19:53,516
MICE.

417
00:19:53,516 --> 00:19:56,118
SO THE QUESTION ARISES, DOES ZAP

418
00:19:56,118 --> 00:20:02,291
IN ADDITION TO EFFECTING VIRAL 

419
00:20:02,291 --> 00:20:04,960
RNAs DOES IT AFFECT HOST RNAs?

420
00:20:04,960 --> 00:20:06,896
USING THESE RULES WHAT WE 

421
00:20:06,896 --> 00:20:08,431
DEVELOPED IS PREDICTION 

422
00:20:08,431 --> 00:20:10,766
ALGORITHM THAT TRIES TO PREDICT 

423
00:20:10,766 --> 00:20:13,836
WHETHER AN INDIVIDUAL TRANSCRIPT

424
00:20:13,836 --> 00:20:15,938
WILL BE SENSITIVE TO IT BASED ON

425
00:20:15,938 --> 00:20:16,305
THESE CRITERIA.

426
00:20:16,305 --> 00:20:20,142
IF YOU COMPARE MAMMALIAN AND 

427
00:20:20,142 --> 00:20:24,246
INSECT TRANSCRIPTOMES ON CGs AND

428
00:20:24,246 --> 00:20:26,048
THE DISTANCE BETWEEN THEM AND 

429
00:20:26,048 --> 00:20:27,917
COMPOSITION BY ALL THREE 

430
00:20:27,917 --> 00:20:30,219
CRITERIA HUMAN TRANSCRIPTS ARE 

431
00:20:30,219 --> 00:20:31,987
PREDICTED TO BE FAR LESS 

432
00:20:31,987 --> 00:20:35,391
SENSITIVE THAN INSECT 

433
00:20:35,391 --> 00:20:38,294
TRANSCRIPTS AND IF YOU PLOT A 

434
00:20:38,294 --> 00:20:40,496
ZAP SENSITIVITY SCORE AN AVERAGE

435
00:20:40,496 --> 00:20:41,564
FLY TRANSCRIPT WOULD BE AS 

436
00:20:41,564 --> 00:20:46,335
SENSITIVE AS THE MOST SENSITIVE 

437
00:20:46,335 --> 00:20:47,837
.1% OF HUMAN TRANSCRIPT.

438
00:20:47,837 --> 00:20:50,439
WE THINK WE CAN PREDICT WHAT 

439
00:20:50,439 --> 00:20:55,010
HUMAN GENES WOULD BE SENSITIVE 

440
00:20:55,010 --> 00:20:55,244
TO ZAP.

441
00:20:55,244 --> 00:20:57,246
AND VERY VERIFIED THAT USING 

442
00:20:57,246 --> 00:20:59,048
KNOCKOUT MICE.

443
00:20:59,048 --> 00:21:01,917
THESE ARE THE HUMAN TRANSCRIPTS 

444
00:21:01,917 --> 00:21:02,985
WITH THE ZAP SENSITIVITY SCORES 

445
00:21:02,985 --> 00:21:05,454
AND WHEN WE MEASURE THEIR LEVELS

446
00:21:05,454 --> 00:21:09,258
IN THE SPLEENS OF ZAP KNOCKOUT 

447
00:21:09,258 --> 00:21:10,960
AND COMPETENT MICE VIRTUALLY ALL

448
00:21:10,960 --> 00:21:13,996
THOSE APPEAR TO BE ELEVATED WHEN

449
00:21:13,996 --> 00:21:16,132
YOU KNOCKOUT ZAP.

450
00:21:16,132 --> 00:21:18,267
THE AVERAGE ZAP SENSITIVITY 

451
00:21:18,267 --> 00:21:21,103
SCORE OF UPREGULATE WILL GENES 

452
00:21:21,103 --> 00:21:22,271
IS HIGHER AS WELL.

453
00:21:22,271 --> 00:21:24,640
SO ZAP REGULATED GENE IS IT 

454
00:21:24,640 --> 00:21:24,874
HIGHER.

455
00:21:24,874 --> 00:21:29,378
THE MAGNITUDE ON HOST GENE 

456
00:21:29,378 --> 00:21:31,313
EXPRESSION IS MODEST BUT BECOMES

457
00:21:31,313 --> 00:21:33,082
EXAGGERATED WHEN YOU INDUCE ZAP 

458
00:21:33,082 --> 00:21:33,783
WITH POLY I.C.

459
00:21:33,783 --> 00:21:35,751
WE DON'T KNOW WHAT THE BIOLOGY 

460
00:21:35,751 --> 00:21:36,852
ASSOCIATED WITH THE GENES ARE 

461
00:21:36,852 --> 00:21:41,991
BUT MANY OF THEM ARE INVOLVED IN

462
00:21:41,991 --> 00:21:43,659
DEPOSITION OF EXTRA CELLULAR 

463
00:21:43,659 --> 00:21:45,895
MATRIX AND THERE'S SECONDARY 

464
00:21:45,895 --> 00:21:47,463
EFFECTS BASED ON TRANSCRIPTION 

465
00:21:47,463 --> 00:21:48,063
FACTORS.

466
00:21:48,063 --> 00:21:51,133
THIS IS BASICALLY A MODEL.

467
00:21:51,133 --> 00:21:56,939
SO AN OPTIMAL TARGET RNA HAS CGs

468
00:21:56,939 --> 00:21:58,674
SPACED THAT AMOUNT A PART AND WE

469
00:21:58,674 --> 00:22:01,977
THINK THIS IS WHY THE ABILITY OF

470
00:22:01,977 --> 00:22:07,483
TRIM 25 TO MAKE MULTIMERS AND 

471
00:22:07,483 --> 00:22:09,185
THE EXTENDED IT WILL ZAP PROTEIN

472
00:22:09,185 --> 00:22:10,119
IS IMPORTANT BECAUSE IT CONVERTS

473
00:22:10,119 --> 00:22:13,422
WHAT'S A MONOVALENT INTERACTION 

474
00:22:13,422 --> 00:22:23,933
TO A MULTIVALENT INTERACTION.

475
00:22:26,302 --> 00:22:29,872
AND OF COURSE CREATES A 

476
00:22:29,872 --> 00:22:30,739
NIGHTMARE FOR STRUCTURAL 

477
00:22:30,739 --> 00:22:31,941
BIOLOGISTS WHO WANT TO STUDY THE

478
00:22:31,941 --> 00:22:32,474
COMPLEX.

479
00:22:32,474 --> 00:22:35,544
THAT'S ALL I HAVE TO SAY.

480
00:22:35,544 --> 00:22:36,612
THE PEOPLE IN MY GROUP WHO 

481
00:22:36,612 --> 00:22:38,214
CONTRIBUTED ARE LISTED THERE BUT

482
00:22:38,214 --> 00:22:46,155
IT'S VERY MUCH A TEAM EFFORT.

483
00:22:46,155 --> 00:22:48,858
AND THERE'S MORE DATA ON A 

484
00:22:48,858 --> 00:22:52,261
POSTER SO I ENCOURAGE YOU TO 

485
00:22:52,261 --> 00:22:54,263
STOP BY.

486
00:22:54,263 --> 00:22:56,332
JENNIFER AND JANET AND MELANIE 

487
00:22:56,332 --> 00:22:57,299
ARE ALREADY CONTRIBUTORS TO 

488
00:22:57,299 --> 00:22:57,533
THIS.

489
00:22:57,533 --> 00:23:07,943
THANK YOU FOR LISTENING.

490
00:23:30,733 --> 00:23:31,100
S 

491
00:23:31,100 --> 00:23:33,535
>> I'D LIKE TO TELL YOU
ABOUT 

492
00:23:33,535 --> 00:23:36,305
OUR EFFORTS TO UNDERSTAND THE 

493
00:23:36,305 --> 00:23:38,040
RNA DETERMINATES OF PACKAGING 

494
00:23:38,040 --> 00:23:40,609
AND IT'S A COLLABORATION WITH 

495
00:23:40,609 --> 00:23:42,878
SEVERAL GROUPS WITHIN THE CRNA.

496
00:23:42,878 --> 00:23:44,880
SO THIS WAS JUST TO REMIND YOU 

497
00:23:44,880 --> 00:23:48,284
THE SINGLE INTEGRATED PROVIRUS 

498
00:23:48,284 --> 00:23:49,351
PRODUCES A FULL LENGTH 

499
00:23:49,351 --> 00:23:50,920
TRANSCRIPT AND THAT HAS TWO 

500
00:23:50,920 --> 00:23:51,520
FUNCTIONS.

501
00:23:51,520 --> 00:23:59,228
IT HAS THE FUNCTION AS AN mRNA 

502
00:23:59,228 --> 00:24:02,097
AND GENOME AND IT'S BEEN THE 

503
00:24:02,097 --> 00:24:04,433
PAST SEVERAL YEARS PEOPLE 

504
00:24:04,433 --> 00:24:05,100
REALIZED ONE OF THE DETERMINATES

505
00:24:05,100 --> 00:24:09,038
OF THE FUNCTIONAL CONTROL IS 

506
00:24:09,038 --> 00:24:11,540
HETEROGENEOUS START SITE USAGE.

507
00:24:11,540 --> 00:24:13,208
FOR MANY YEARS IT WAS UNKNOWN 

508
00:24:13,208 --> 00:24:18,213
WHAT THE START SITE IS.

509
00:24:18,213 --> 00:24:20,316
THESE ARE THE THREE GUANOSINES 

510
00:24:20,316 --> 00:24:21,784
THAT COULD START AS A SITE BUT 

511
00:24:21,784 --> 00:24:32,261
IT'S NOT PEOPLE DIDN'T CARE.

512
00:24:34,363 --> 00:24:38,567
AND THERE'S 1G AND IN 2015 THEY 

513
00:24:38,567 --> 00:24:40,602
SUGGESTED ALL THREE ARE MADE AND

514
00:24:40,602 --> 00:24:44,974
3G IS THE PREDOMINANT ONE.

515
00:24:44,974 --> 00:24:46,942
IF YOU GO TO THE NIH COMPENDIUM 

516
00:24:46,942 --> 00:24:49,011
ALL THREE ARE REPORTED BUT 

517
00:24:49,011 --> 00:24:51,046
THEY'RE SPECULATION ABOUT WHAT 

518
00:24:51,046 --> 00:24:52,014
THEY WERE.

519
00:24:52,014 --> 00:24:54,616
WITH ALICE AND SERGEI WHAT WE 

520
00:24:54,616 --> 00:24:57,553
SHOWED IN 2016 IS THAT ALL THREE

521
00:24:57,553 --> 00:25:00,622
ARE TRANSCRIBED IN CELLS THAT 

522
00:25:00,622 --> 00:25:05,728
THE 1G FORM IS ENRICHED IN 

523
00:25:05,728 --> 00:25:08,564
VARIANTS AND WHAT THEY SHOWED 

524
00:25:08,564 --> 00:25:12,134
THE IT 3G FORM IS ENRICHED IN 

525
00:25:12,134 --> 00:25:15,404
POLYSOMES AND THE 13 AND 3G ARE 

526
00:25:15,404 --> 00:25:16,872
PREDOMINANT AND SEEN IN HUMAN 

527
00:25:16,872 --> 00:25:18,607
GENES AS WELL AND IT'S BEEN 

528
00:25:18,607 --> 00:25:20,476
CALLED TWIN TRANSCRIPTIONAL 

529
00:25:20,476 --> 00:25:21,410
START SITE USAGE.

530
00:25:21,410 --> 00:25:26,148
WHAT LEADS TO THIS PARADIGM 

531
00:25:26,148 --> 00:25:28,784
WHERE THE 3G CAPPED RNAs AND ONE

532
00:25:28,784 --> 00:25:31,820
AS A GENOME AND NOW THIS HAS 

533
00:25:31,820 --> 00:25:33,555
BEEN SPORTED BY A NUMBER OF 

534
00:25:33,555 --> 00:25:35,824
ADDITIONAL STUDIES AND TWO THAT 

535
00:25:35,824 --> 00:25:38,794
STAND OUT IN MY MIND ARE ALICE'S

536
00:25:38,794 --> 00:25:42,297
GROUP THAT SHOWED THE 3G FORM IS

537
00:25:42,297 --> 00:25:44,366
ENRICHED IN SPLICE TRANSCRIPTS 

538
00:25:44,366 --> 00:25:46,301
AND ALSO WILL ANOTHER GROUP 

539
00:25:46,301 --> 00:25:48,637
SHOWED THE 3G ONLY VIRUS HAS A 

540
00:25:48,637 --> 00:25:49,838
PACKAGING DEFECT WHICH WOULD 

541
00:25:49,838 --> 00:25:52,207
MAKE SENSE BASED ON THE MODEL 

542
00:25:52,207 --> 00:25:54,710
AND THE 1G VIRUS IS REDUCED IN 

543
00:25:54,710 --> 00:25:56,378
GAG EXPRESSION WHICH ALSO MAKES 

544
00:25:56,378 --> 00:25:59,181
SENSE WITH THE PARADIGM.

545
00:25:59,181 --> 00:26:04,053
WE'VE USED NMR TO UNDERSTAND 

546
00:26:04,053 --> 00:26:06,188
STRUCTURALLY HOW IT WORKS AND 

547
00:26:06,188 --> 00:26:10,125
FOR THE IT WILL VERSION OF HIV 

548
00:26:10,125 --> 00:26:13,095
WE SHOWED THE FIVE 2G AND 3G 

549
00:26:13,095 --> 00:26:13,996
RNAs FORM A STRUCTURE LIKE THIS.

550
00:26:13,996 --> 00:26:16,398
THIS IS JUST A CARTOON BUT 

551
00:26:16,398 --> 00:26:20,836
RESIDUES THAT HAD BEEN PREDICTED

552
00:26:20,836 --> 00:26:22,604
TO FORM A HAIRPIN OR POLY A 

553
00:26:22,604 --> 00:26:27,576
REGION THIS FORM OF THE RNA 

554
00:26:27,576 --> 00:26:29,578
EXPOSES THE SPLICED DONOR SITE 

555
00:26:29,578 --> 00:26:32,648
ON THE NUKE LEO CAPSID BINDING 

556
00:26:32,648 --> 00:26:35,551
SITES ARE SEQUESTERED AND THIS 

557
00:26:35,551 --> 00:26:38,854
FUNCTIONS AS mRNA AND THIS IS 

558
00:26:38,854 --> 00:26:40,823
WORK OF JOSH BROWN SHOWED THAT 

559
00:26:40,823 --> 00:26:45,794
IF YOU HAVE THE KAPPA 1G FORM 

560
00:26:45,794 --> 00:26:47,196
THESE STACK END TO END.

561
00:26:47,196 --> 00:26:49,698
WHAT THAT DOES IS IT SEQUESTERS 

562
00:26:49,698 --> 00:26:52,768
THE CAP IN THIS STRUCTURE THE 

563
00:26:52,768 --> 00:26:54,436
SPLICE DONOR SITE AND THE GAG 

564
00:26:54,436 --> 00:26:57,573
START CODE ARE ALSO SEQUESTERED.

565
00:26:57,573 --> 00:27:04,446
NUKE NUCLEOBINDING SITES ARE 

566
00:27:04,446 --> 00:27:05,914
EXPOSED AND THIS IS SEQUESTERED 

567
00:27:05,914 --> 00:27:07,116
AND THIS IS THE FORM THAT 

568
00:27:07,116 --> 00:27:09,384
FUNCTIONS AS THE GENOME.

569
00:27:09,384 --> 00:27:13,222
SO WE THINK WE UNDERSTAND THE 

570
00:27:13,222 --> 00:27:15,390
STRUCTURAL DETERMINATES OF RNA 

571
00:27:15,390 --> 00:27:15,757
PACKAGING.

572
00:27:15,757 --> 00:27:17,826
THE QUESTION IS DOES 

573
00:27:17,826 --> 00:27:19,528
TRANSCRIPTIONAL START SITE 

574
00:27:19,528 --> 00:27:20,529
HETEROGENEITY INFLUENCE 

575
00:27:20,529 --> 00:27:22,030
TRANSLATION AND WHAT IS THE ROLE

576
00:27:22,030 --> 00:27:31,573
OF POLY A AND THIS WAS STUDIED.

577
00:27:31,573 --> 00:27:35,010
SO THIS IS INTERESTING BECAUSE 

578
00:27:35,010 --> 00:27:36,645
YOU'LL NOTICE IT'S THE POLY A 

579
00:27:36,645 --> 00:27:39,181
ELEMENT THAT INTERACTS WITH THE 

580
00:27:39,181 --> 00:27:41,917
ADDITIONAL GUANOSINE.

581
00:27:41,917 --> 00:27:44,153
NO OTHER MMR STRUCTURE WE DIDN'T

582
00:27:44,153 --> 00:27:47,256
SEE INTERACTIONS GOING FROM THE 

583
00:27:47,256 --> 00:27:49,324
1G TO 3G.

584
00:27:49,324 --> 00:27:50,926
THE POLY A ELEMENT PLAYS AN 

585
00:27:50,926 --> 00:27:51,593
IMPORTANT ROLE.

586
00:27:51,593 --> 00:27:54,263
OUR WORK IS BASED ON ACTUAL 

587
00:27:54,263 --> 00:27:56,198
OBSERVABLE INTERACTIONS.

588
00:27:56,198 --> 00:28:00,602
THIS IS THE OBSERVED SOLVED OF 

589
00:28:00,602 --> 00:28:03,572
NMR AND WHAT IS BOXED ARE 

590
00:28:03,572 --> 00:28:05,040
EXPERIMENTALLY DETERMINED NOT 

591
00:28:05,040 --> 00:28:06,675
PREDICTED BASED ON FREE ENERGY 

592
00:28:06,675 --> 00:28:10,112
CALCULATIONS AN IT'S CLEAR POLY 

593
00:28:10,112 --> 00:28:11,747
A IS NOT MAKING LONG RANGE 

594
00:28:11,747 --> 00:28:12,080
INTERACTIONS.

595
00:28:12,080 --> 00:28:14,583
I THINK IT'S WORTH POINTING OUT 

596
00:28:14,583 --> 00:28:18,287
THERE ARE SEVERAL OTHER MODELS 

597
00:28:18,287 --> 00:28:27,095
OF THE MONOMERRICK -- MONOMERIC 

598
00:28:27,095 --> 00:28:29,398
AND WE OBSERVED THESE 

599
00:28:29,398 --> 00:28:30,566
INTERACTIONS BUT DIDN'T LOOK FOR

600
00:28:30,566 --> 00:28:31,833
OTHER INTERACTIONS.

601
00:28:31,833 --> 00:28:35,237
IN FACT WE BELIEVE NOW THAT 

602
00:28:35,237 --> 00:28:36,672
THESE PROBABLY ADOPT THE SAME 

603
00:28:36,672 --> 00:28:38,440
STRUCTURE THOUGH THIS STILL A 

604
00:28:38,440 --> 00:28:40,909
PREDICTION ON OUR PART.

605
00:28:40,909 --> 00:28:41,577
THERE ARE OTHER STRUCTURES 

606
00:28:41,577 --> 00:28:43,845
SOLVED OR PROPOSED ON THE BASIS 

607
00:28:43,845 --> 00:28:48,016
OF CHEMICAL PROBING.

608
00:28:48,016 --> 00:28:51,587
THIS IN 2003 BY BERKEHOUT AND 

609
00:28:51,587 --> 00:28:53,021
THERE'S INTERACTION WITH THE 

610
00:28:53,021 --> 00:28:55,457
DIMER INITIATION SITE AND 

611
00:28:55,457 --> 00:28:56,692
SHIFTED A BIT BUT THERE'S DOWN 

612
00:28:56,692 --> 00:28:58,393
STREAM INTERACTIONS WE DID NOT 

613
00:28:58,393 --> 00:28:59,795
SEE.

614
00:28:59,795 --> 00:29:02,064
SIMILAR TO ANOTHER RECENT MODEL 

615
00:29:02,064 --> 00:29:03,732
THAT MIKE AND HIS LAB PROPOSED 

616
00:29:03,732 --> 00:29:05,500
BASED ON CHEMICAL PROBING.

617
00:29:05,500 --> 00:29:08,804
I SHOULD ALSO POINT OUT THEY'RE 

618
00:29:08,804 --> 00:29:09,404
DIFFERENT SIZES.

619
00:29:09,404 --> 00:29:11,740
THESE ARE RNA'S THROUGH THE GAG 

620
00:29:11,740 --> 00:29:14,109
START CODE AND THESE ARE 

621
00:29:14,109 --> 00:29:15,611
SOMEWHAT LARGER RNAs.

622
00:29:15,611 --> 00:29:19,481
ONE IS IN THE MIDDLE BY THE 

623
00:29:19,481 --> 00:29:20,382
GROUP REPORTED.

624
00:29:20,382 --> 00:29:21,583
THIS IS QUITE A BIT DIFFERENT 

625
00:29:21,583 --> 00:29:22,184
FROM ALL THESE OTHER PROPOSED 

626
00:29:22,184 --> 00:29:25,053
MODELS.

627
00:29:25,053 --> 00:29:26,288
THE COMMON FEATURE IS THEY ALL 

628
00:29:26,288 --> 00:29:30,292
HAVE AN EXTENDED POLY A ELEMENT 

629
00:29:30,292 --> 00:29:39,101
THAT IS NOT A HAIRPIN IN THE 

630
00:29:39,101 --> 00:29:39,434
MO

631
00:29:39,434 --> 00:29:40,969
MONOMER FORM AND WANTED TO SEE 

632
00:29:40,969 --> 00:29:42,804
IF THE STRUCTURE IS CONSERVED.

633
00:29:42,804 --> 00:29:48,176
WE DID AN ANALYSIS OF ALL THE 

634
00:29:48,176 --> 00:29:58,720
MAJOR HIV SUB TYPES IN THE LOS 

635
00:30:03,492 --> 00:30:06,295
ALOMOS COMPENDIUM AND THERE'S 

636
00:30:06,295 --> 00:30:08,930
ENERGY OF THE HAIRPIN FORM POLY 

637
00:30:08,930 --> 00:30:12,534
A AND HAVE DIFFERENT LOCATIONS 

638
00:30:12,534 --> 00:30:18,640
OF BULGES AND NON-CANONICAL BASE

639
00:30:18,640 --> 00:30:20,509
PAIRS BUT CONSERVED THROUGH THE 

640
00:30:20,509 --> 00:30:20,776
SEQUENCES.

641
00:30:20,776 --> 00:30:23,478
WE MADE THE POLY A AND DELETED 

642
00:30:23,478 --> 00:30:26,281
THESE AND CALL THIS AN IDEALIZED

643
00:30:26,281 --> 00:30:28,083
HELIX AND CALLED IT AN AU AND 

644
00:30:28,083 --> 00:30:30,886
GOT RID OF THE BULGES AND LED TO

645
00:30:30,886 --> 00:30:33,822
A 15 DEGREE STABILIZATION.

646
00:30:33,822 --> 00:30:36,992
WE WOULD CALL THIS A POLY A 

647
00:30:36,992 --> 00:30:39,628
HAIRPIN WITH NO BULGES OR 

648
00:30:39,628 --> 00:30:40,696
IDEALIZED HAIRPIN.

649
00:30:40,696 --> 00:30:44,032
WE MADE THESE SAME MUTATIONS IN 

650
00:30:44,032 --> 00:30:48,303
THE INTACT LEADER IN THE NMR 

651
00:30:48,303 --> 00:30:50,072
SHOWED WILL POLY A IS A HAIRPIN 

652
00:30:50,072 --> 00:30:51,640
IN THE INTACT LEADER AND THIS 

653
00:30:51,640 --> 00:30:54,009
HAS THREE GUANOSINES IN THE FIVE

654
00:30:54,009 --> 00:30:55,277
PRIME END WHERE IT WOULD 

655
00:30:55,277 --> 00:30:59,014
NORMALLY NOT BE A HAIRPIN.

656
00:30:59,014 --> 00:31:02,751
THIS FORM BINDS NUCLEOCAPSID 

657
00:31:02,751 --> 00:31:11,460
LIKE THE DIMERIC FORM AND THIS 

658
00:31:11,460 --> 00:31:13,595
IS A MODEL FOR THE CAP 3G AND 

659
00:31:13,595 --> 00:31:17,265
WHEN HAVE YOU MORE Gs THE WILD 

660
00:31:17,265 --> 00:31:27,809
TYPE IS MONOMERRICK -- MONOMERIC

661
00:31:28,143 --> 00:31:34,516
IT'S A DIMER LONG AS POLY A IS A

662
00:31:34,516 --> 00:31:44,693
HAIR PIN.

663
00:31:46,361 --> 00:31:49,531
AND THIS FORMS A DIMER AND THAT 

664
00:31:49,531 --> 00:31:50,866
BINDS NUKE LEE CAPSID IT STILL 

665
00:31:50,866 --> 00:31:57,706
HAS AN EXPOSED CAP.

666
00:31:57,706 --> 00:32:01,476
AND WE HAVE THE HAIRPIN FOR THE 

667
00:32:01,476 --> 00:32:03,578
POLY A THOUGH THERE'S AN EXPOSED

668
00:32:03,578 --> 00:32:07,582
CAP AND THOUGH IT DIMERIZES IN 

669
00:32:07,582 --> 00:32:07,949
CAPSID.

670
00:32:07,949 --> 00:32:09,985
SO WITH ALICE AND SERGEI THEY 

671
00:32:09,985 --> 00:32:12,187
ARE SHOWED IF YOU TAKE AND LOOK 

672
00:32:12,187 --> 00:32:16,158
AT THE MAL STRAIN IN CELLS IF 

673
00:32:16,158 --> 00:32:20,662
THE TWO RNA'S ARE MADE AT 

674
00:32:20,662 --> 00:32:23,899
SIMILAR LEVELS ONLY THE CAP 1G 

675
00:32:23,899 --> 00:32:25,200
FORM GETS INTO VIRUSES AND SAME 

676
00:32:25,200 --> 00:32:27,636
FOR THE NL43 AND THEY'RE BOTH 

677
00:32:27,636 --> 00:32:30,972
MADE INDEPENDENTLY IN CELLS BUT 

678
00:32:30,972 --> 00:32:33,442
ONLY THE 1G IS COMPETITIVELY 

679
00:32:33,442 --> 00:32:34,810
PACKAGE SOD THE 3G THOUGH IT HAS

680
00:32:34,810 --> 00:32:40,415
ALL THE PROTEIN BINDING SITE IT 

681
00:32:40,415 --> 00:32:43,218
DIMERIZES AND NUCLEOCAPSID 

682
00:32:43,218 --> 00:32:44,686
BINDING ARE NOT SUFFICIENT TO 

683
00:32:44,686 --> 00:32:46,121
OVERCOME THE DOMINANT NEGATIVE 

684
00:32:46,121 --> 00:32:48,390
EFFECT OF CAP EXPOSURE.

685
00:32:48,390 --> 00:32:50,025
AND AS CONSISTENT WITH PRIOR 

686
00:32:50,025 --> 00:32:53,061
FINDINGS WE PUBLISHED IF WE MADE

687
00:32:53,061 --> 00:32:54,796
THIS CORE SIGNAL THAT GETS 

688
00:32:54,796 --> 00:32:57,532
CAPPED IN CELLS NOT PACKAGED BUT

689
00:32:57,532 --> 00:33:04,072
IF WE ADD AN RIBOZYME THIS IS 

690
00:33:04,072 --> 00:33:09,911
COMPETITIVELY PACKAGED AND ADD A

691
00:33:09,911 --> 00:33:20,455
FEW RESIDUES ON TO THE DIMERIC 

692
00:33:26,328 --> 00:33:29,164
PACKAGING AND WHAT ABOUT 

693
00:33:29,164 --> 00:33:32,334
FUNCTIONS THAT COULD BE 

694
00:33:32,334 --> 00:33:34,169
INFLUENCED BY HETEROGENOUS START

695
00:33:34,169 --> 00:33:35,103
SITE USAGE?

696
00:33:35,103 --> 00:33:40,642
WE ENTERED INTO A COLLABORATION 

697
00:33:40,642 --> 00:33:42,310
WITH THE SHEER LAB AND THEY 

698
00:33:42,310 --> 00:33:44,412
PLAYED A HUGE ROLE IN THIS WORK 

699
00:33:44,412 --> 00:33:48,083
AND THESE ARE TWO VECTORS THAT 

700
00:33:48,083 --> 00:33:50,252
WOULD BE CO-TRAN SPECTED INTO 

701
00:33:50,252 --> 00:33:52,954
CELLS AND ENCODE FOR RNAs WITH 

702
00:33:52,954 --> 00:33:53,788
DIFFERENT COLORS.

703
00:33:53,788 --> 00:33:56,558
THIS IS THE YFP AND CFP.

704
00:33:56,558 --> 00:34:00,529
THEN WHAT WE CAN DO IS PUT 

705
00:34:00,529 --> 00:34:02,264
DIFFERENT TRANSCRIPTION START 

706
00:34:02,264 --> 00:34:04,566
SITES BASED ON THE WORK OF ALICE

707
00:34:04,566 --> 00:34:05,567
AND SERGEI.

708
00:34:05,567 --> 00:34:11,239
NOW WE CAN HAVE LET'S SAY A 

709
00:34:11,239 --> 00:34:16,511
YELLOW RNA BEGINS WITH 1G AND 

710
00:34:16,511 --> 00:34:18,246
CYAN RNA WITH 3G.

711
00:34:18,246 --> 00:34:20,148
HERE'S AN EXAMPLE ON THE LEFT 

712
00:34:20,148 --> 00:34:25,554
WITH A CYAN COLORED AND YELLOW 

713
00:34:25,554 --> 00:34:26,888
COLORED 1G.

714
00:34:26,888 --> 00:34:30,191
YOU CAN SEE THE CYAN IS GROWING 

715
00:34:30,191 --> 00:34:33,028
AND MORE INTENSE.

716
00:34:33,028 --> 00:34:34,195
IT LOOKS A LITTLE BIT BETTER 

717
00:34:34,195 --> 00:34:38,300
WHERE WE REVERSE THE COLORS.

718
00:34:38,300 --> 00:34:39,467
AND IT'S FWROEG MORE QUICKLY AND

719
00:34:39,467 --> 00:34:43,271
MORE INTENSE THAN THE CYAN 

720
00:34:43,271 --> 00:34:46,274
SHOWING IT'S THE 3G FORM THAT IS

721
00:34:46,274 --> 00:34:49,778
MORE EFFICIENTLY TRANSLATED IN 

722
00:34:49,778 --> 00:34:52,080
CELLS IN THIS COMPETITIVE ASSAY 

723
00:34:52,080 --> 00:34:53,448
AND PLOT THESE AND PICK REGIONS 

724
00:34:53,448 --> 00:34:56,351
WHERE WE CAN DO COMPARISONS.

725
00:34:56,351 --> 00:35:00,155
SO WHAT WE NOW KNOW IN OUR 

726
00:35:00,155 --> 00:35:02,357
CONTROLS IF WE COMPARE 1G AND 3G

727
00:35:02,357 --> 00:35:06,294
AND THEN OUR 3G NO BULGE IF THE 

728
00:35:06,294 --> 00:35:13,001
TWO CONTROLS HAVE THE SAME FIVE 

729
00:35:13,001 --> 00:35:17,339
PRIECHL  -- PRIME END AND 

730
00:35:17,339 --> 00:35:20,508
TRANSLATED WELL AND LOOK AT 1G 

731
00:35:20,508 --> 00:35:21,676
VERSUS 3G THE 3G IS MORE 

732
00:35:21,676 --> 00:35:25,146
EFFICIENTLY TRANSLATED AND NOW 

733
00:35:25,146 --> 00:35:27,682
BY REVERSING THESE TO SHOW AS A 

734
00:35:27,682 --> 00:35:30,318
CONTROL IT'S ALWAYS THE 3G MORE 

735
00:35:30,318 --> 00:35:30,819
EFFICIENTLY TRANSLATED.

736
00:35:30,819 --> 00:35:34,122
IF WE LOOK AT THE 3G WITH A NO 

737
00:35:34,122 --> 00:35:38,159
BULGE YOU CAN SEE THIS IS NOT AS

738
00:35:38,159 --> 00:35:40,962
EFFICIENTLY TRANSLATED IS MORE 

739
00:35:40,962 --> 00:35:42,998
EFFICIENTLY TRANSLATED THAN THE 

740
00:35:42,998 --> 00:35:46,201
1G AND THEN IN FACT THE 3G WITH 

741
00:35:46,201 --> 00:35:49,070
NO BULGE AND THE 3G ARE 

742
00:35:49,070 --> 00:35:50,472
TRANSLATED ABOUT EQUALLY WELL.

743
00:35:50,472 --> 00:35:53,041
WHAT DOES THIS ALL MEAN?

744
00:35:53,041 --> 00:35:55,810
WHAT IT MEANS IS THAT IF YOU 

745
00:35:55,810 --> 00:35:58,279
HAVE THE WILD TYPE RNA THAT HAS 

746
00:35:58,279 --> 00:36:01,216
A POLY A AND HAIRPIN FORM THE 

747
00:36:01,216 --> 00:36:02,751
FIVE PRIME CAP IS SEQUESTERED 

748
00:36:02,751 --> 00:36:04,753
AND THIS LIKES THE DIMERIZED.

749
00:36:04,753 --> 00:36:09,324
IT HAS HIGH AFFINITY NUKE LEO 

750
00:36:09,324 --> 00:36:09,858
CAPSID BINDING SITE AND 

751
00:36:09,858 --> 00:36:12,227
TRANSLATION EFFICIENCY IS LOW.

752
00:36:12,227 --> 00:36:13,962
IF YOU HAVE THE 3G FORM OF THE 

753
00:36:13,962 --> 00:36:17,232
RNA, THIS HAS AN EXTENDED POLY A

754
00:36:17,232 --> 00:36:21,836
AND EXPOSED FIVE PRIME CAP AND 

755
00:36:21,836 --> 00:36:32,380
THE MONOMER AND THIS HAS ALL THE

756
00:36:40,221 --> 00:36:42,290
PROTEIN BINDING SITES BUT 

757
00:36:42,290 --> 00:36:44,559
PACKAGING EFFICIENCY IS LOW 

758
00:36:44,559 --> 00:36:46,461
BECAUSE THE CAP IS EXPOSED AND 

759
00:36:46,461 --> 00:36:50,131
TRANSLATION EFFICIENCY IS HIGH.

760
00:36:50,131 --> 00:36:52,434
WE DON'T JUST THINK IT'S A POLY 

761
00:36:52,434 --> 00:36:54,302
A SWITCH BECAUSE IT'S NOT THE 

762
00:36:54,302 --> 00:37:00,275
ONLY REGION THAT UNDER GOES 

763
00:37:00,275 --> 00:37:01,376
CHANGES AND WE'RE COMPARING THE 

764
00:37:01,376 --> 00:37:04,813
FREE ENERGY CALCULATED FOR THE 

765
00:37:04,813 --> 00:37:06,014
WILD TYPE SEQUENCE VERSUS THE 

766
00:37:06,014 --> 00:37:08,950
ENERGY OF A MUTATED FORM THAT 

767
00:37:08,950 --> 00:37:11,219
WOULD BE AN IDEALIZED HAIRPIN 

768
00:37:11,219 --> 00:37:12,454
AND HELIX.

769
00:37:12,454 --> 00:37:16,925
YOU CAN SEE THESE ALL HAVE 

770
00:37:16,925 --> 00:37:18,293
HIGHLY SUPPRESSED FREE ENERGIES 

771
00:37:18,293 --> 00:37:19,394
OF FOLDING.

772
00:37:19,394 --> 00:37:21,930
SO WE THINK THESE DEPRESSED 

773
00:37:21,930 --> 00:37:23,298
FOLDING FREE ENERGIES ARE 

774
00:37:23,298 --> 00:37:26,267
IMPORTANT FOR ALLOWING THE RNA 

775
00:37:26,267 --> 00:37:29,637
TO ADOPT MULTIPLE CONFIRMATIONS.

776
00:37:29,637 --> 00:37:31,906
THE ENTIRE LEADER IS 

777
00:37:31,906 --> 00:37:35,744
ENERGETICALLY TUNED TO ENABLE 

778
00:37:35,744 --> 00:37:36,778
POLYMORPHISM.

779
00:37:36,778 --> 00:37:39,681
TO CONCLUDE IN TERMS OF 

780
00:37:39,681 --> 00:37:42,283
PACKAGING DIMERIZATION EXPOSES 

781
00:37:42,283 --> 00:37:45,687
BINDING SITES AND IT'S A 

782
00:37:45,687 --> 00:37:50,125
DOMINANT NEGATIVE DETERMINANT OF

783
00:37:50,125 --> 00:37:50,492
PACKAGING.

784
00:37:50,492 --> 00:37:52,794
IF YOU WANT TO GET RNAs TO 

785
00:37:52,794 --> 00:37:54,529
VIRUSES YOU HAVE TO EXPOSE THESE

786
00:37:54,529 --> 00:37:56,865
PACKAGING SIGNALS AND SEQUESTER 

787
00:37:56,865 --> 00:37:57,298
THE CAP.

788
00:37:57,298 --> 00:37:59,667
IN TERMS OF TRANSLATION CAP 

789
00:37:59,667 --> 00:38:03,671
EXPOSURE IS A POSITIVE EFFECTER 

790
00:38:03,671 --> 00:38:04,539
AT EFFICIENT TRANSLATION.

791
00:38:04,539 --> 00:38:08,977
THE WORK IN MY LAB STARTED BY 

792
00:38:08,977 --> 00:38:10,011
JOSH BROWN NOW A PHYSICIAN 

793
00:38:10,011 --> 00:38:11,479
SCIENTIST AT EMORY BUT ALL THE 

794
00:38:11,479 --> 00:38:16,818
WORK I TALKED ABOUT WAS CARRIED 

795
00:38:16,818 --> 00:38:18,486
OUT BY A CURRENT Ph.D. STUDENT 

796
00:38:18,486 --> 00:38:18,753
IN MY LAB.

797
00:38:18,753 --> 00:38:21,689
THANKS.

798
00:38:21,689 --> 00:38:21,756
 

799
00:38:47,782 --> 00:38:49,851
>> OKAY.

800
00:38:49,851 --> 00:38:51,619
SO I'M GOING TO TELL YOU A 

801
00:38:51,619 --> 00:38:53,188
LITTLE BIT OUR TAKE ON TRYING TO

802
00:38:53,188 --> 00:38:54,088
UNDERSTAND WHAT'S HAPPENING 

803
00:38:54,088 --> 00:38:55,623
DURING THE TRANSCRIPTION 

804
00:38:55,623 --> 00:39:01,963
INITIATION COMPLEX FORMATION.

805
00:39:01,963 --> 00:39:03,665
ALL THIS WORK HAS BEEN DONE BY 

806
00:39:03,665 --> 00:39:09,671
THESE TALENTED INDIVIDUALS AND 

807
00:39:09,671 --> 00:39:12,740
COLLABORATION WITH THE LABS AND 

808
00:39:12,740 --> 00:39:14,275
ZOOM THROUGH THE WORK I 

809
00:39:14,275 --> 00:39:17,579
PRESENTED A FEW YEARS AGO HERE 

810
00:39:17,579 --> 00:39:19,480
AND CONCENTRATE ON SOME OF THE 

811
00:39:19,480 --> 00:39:21,049
RD STRUCTURES WE'RE SEEING RIGHT

812
00:39:21,049 --> 00:39:21,449
NOW.

813
00:39:21,449 --> 00:39:24,519
SO AS A QUICK INTRODUCTION FOR 

814
00:39:24,519 --> 00:39:28,656
PEOPLE WHO DON'T THINK ABOUT 

815
00:39:28,656 --> 00:39:32,594
TRAN SKRINGS, AND THERE'S A 

816
00:39:32,594 --> 00:39:37,699
SITE -- TRANSCRIPTION AND 

817
00:39:37,699 --> 00:39:46,374
THERE'S A SITE WITH A NUCLEOTIDE

818
00:39:46,374 --> 00:39:47,876
COMPLIMENTARITY AND THIS IS WHAT

819
00:39:47,876 --> 00:39:50,445
IT RECOGNIZES AND STARTS MAKING 

820
00:39:50,445 --> 00:39:53,081
DNA.

821
00:39:53,081 --> 00:39:57,218
AND WE STARTED SOLVING THE 

822
00:39:57,218 --> 00:39:58,286
STRUCTURE OF THE RNA COMPETENCE 

823
00:39:58,286 --> 00:40:00,622
AND I WON'T SHOW YOU STRUCTURES.

824
00:40:00,622 --> 00:40:03,558
I'LL DO THIS ALL BY CARTOON.

825
00:40:03,558 --> 00:40:09,697
SO IF YOU LOOK AT THE U5PBS IT 

826
00:40:09,697 --> 00:40:12,567
HAS HELIX 2 AND 1 AND AS THE A 

827
00:40:12,567 --> 00:40:18,206
LINK THAT WAS HARD TO FIGURE OUT

828
00:40:18,206 --> 00:40:23,878
THE FOLD BUT IN THE NMR IT FOLDS

829
00:40:23,878 --> 00:40:26,281
BEAUTIFULLY INTO THE MODULES AND

830
00:40:26,281 --> 00:40:30,451
THEY'RE CLOSED BY A SINGLE BASE 

831
00:40:30,451 --> 00:40:30,652
PAIR.

832
00:40:30,652 --> 00:40:33,321
ONCE THE TRNA COMES IN I'LL 

833
00:40:33,321 --> 00:40:35,757
COLOR CODE THIS DIFFERENTLY.

834
00:40:35,757 --> 00:40:39,360
THE TRNA COMES IN AND HELIX 1 

835
00:40:39,360 --> 00:40:40,328
OPENS UP AND GET TWO OTHER 

836
00:40:40,328 --> 00:40:48,636
MODULE FORMATIONS.

837
00:40:48,636 --> 00:40:50,972
AND THIS IS KIND OF SPECIAL AND 

838
00:40:50,972 --> 00:41:01,482
THE M5 OVER HERE IS A PART OF 

839
00:41:02,350 --> 00:41:08,056
THE TRN AND THE NUCLEOTIDES.

840
00:41:08,056 --> 00:41:11,125
WHAT WE KNOW ABOUT IT HELPS RNA 

841
00:41:11,125 --> 00:41:13,394
STRUCTURES FORM BUT CAN ALSO DO 

842
00:41:13,394 --> 00:41:18,299
SOMETHING CALLED LONG RANGE 

843
00:41:18,299 --> 00:41:19,834
DOCKING INTERACTIONS.

844
00:41:19,834 --> 00:41:21,569
AND USING BASE PAIR STACKS AND 

845
00:41:21,569 --> 00:41:23,371
THAT'S WHAT'S HAPPENING HERE.

846
00:41:23,371 --> 00:41:28,509
THE M1 DOCKS ON TO THE NEWLY 

847
00:41:28,509 --> 00:41:33,114
FORMED STEM OF THE RNA AND TO AN

848
00:41:33,114 --> 00:41:36,217
M3 DOCKS AND 18 BASE PAIR PRIMER

849
00:41:36,217 --> 00:41:39,721
AND THE FIRST GA THAT IS GOING 

850
00:41:39,721 --> 00:41:41,990
TO BE THE INITIATION SITE.

851
00:41:41,990 --> 00:41:44,826
TO US IT LOOKS LIKE A HIDDEN 

852
00:41:44,826 --> 00:41:45,727
COMPLEX REPRESSED AND SOMETHING 

853
00:41:45,727 --> 00:41:50,898
THAT NEEDS TO BE OPENED UP TO 

854
00:41:50,898 --> 00:41:51,366
RECOGNIZE IT.

855
00:41:51,366 --> 00:41:57,705
I'M GOING TO QUICKLY DO THIS 

856
00:41:57,705 --> 00:42:04,879
AGAIN AND THIS IS HAPPENING.

857
00:42:04,879 --> 00:42:07,782
YOU HAVE THIS STRUCTURE IT AND 

858
00:42:07,782 --> 00:42:12,453
M2 AND M3 ARE SEQUESTERING THE 

859
00:42:12,453 --> 00:42:14,188
NUCLEOTIDE PRIMER AND 

860
00:42:14,188 --> 00:42:15,690
SEQUESTERING THE FIRST 

861
00:42:15,690 --> 00:42:16,591
INITIATION NUCLEOTIDE.

862
00:42:16,591 --> 00:42:18,960
WE INITIALLY THOUGHT IT WAS 

863
00:42:18,960 --> 00:42:22,997
GOING TO BE THE NUCLEIC ACID 

864
00:42:22,997 --> 00:42:25,833
PROTEIN THAT RIVALLED THIS BUT 

865
00:42:25,833 --> 00:42:27,869
THE R2 BOUND TO THE HELIX AND 

866
00:42:27,869 --> 00:42:31,039
THIS IS A HIGH AFFINITY BINDING 

867
00:42:31,039 --> 00:42:32,840
AND THE JUNCTION GETS REMODULED 

868
00:42:32,840 --> 00:42:38,046
AND THE M2, M3, M4 ALL COME OFF 

869
00:42:38,046 --> 00:42:39,714
AND THE SECOND RT BINDS AND YOU 

870
00:42:39,714 --> 00:42:41,082
CAN GET INITIATION.

871
00:42:41,082 --> 00:42:46,254
SO WE'RE CALLING THIS THE 

872
00:42:46,254 --> 00:42:48,489
REMODELER IT.

873
00:42:48,489 --> 00:42:49,757
SOME MODULES ARE CONSERVED AND 

874
00:42:49,757 --> 00:42:52,960
SOME STRAINS ARE INSERTED TWO 

875
00:42:52,960 --> 00:42:54,996
OTHER MODULES.

876
00:42:54,996 --> 00:42:57,498
THEY'RE VERY IMPORTANT FOR 

877
00:42:57,498 --> 00:42:57,999
INITIATIONS.

878
00:42:57,999 --> 00:43:01,369
EVEN IF YOU MAKE MUTATION DON'T 

879
00:43:01,369 --> 00:43:03,604
ALLOW THE ARCHITECTURE THIS IS 

880
00:43:03,604 --> 00:43:06,941
WORK FROM ALICE'S LAB AND SEE 

881
00:43:06,941 --> 00:43:07,742
THE INITIATION SITE THAT'S 

882
00:43:07,742 --> 00:43:11,979
SUPPOSED TO START AND IT GETS 

883
00:43:11,979 --> 00:43:15,616
VERY MESSED UP EVEN BY TINY 

884
00:43:15,616 --> 00:43:15,950
MANIPULATIONS.

885
00:43:15,950 --> 00:43:17,785
IN VITRO LET'S DID AN EXPERIMENT

886
00:43:17,785 --> 00:43:19,854
WHERE YOU TAKE THE FOUR DOCKING 

887
00:43:19,854 --> 00:43:21,522
INTERACTIONS OUT AND DELETE THEM

888
00:43:21,522 --> 00:43:24,192
AND YOU WOULD THINK IT SHOULD BE

889
00:43:24,192 --> 00:43:26,360
ABLE TO BIND HERE AND IT 

890
00:43:26,360 --> 00:43:29,363
INITIATES ACCURATELY BUT IT 

891
00:43:29,363 --> 00:43:30,064
DOESN'T HAPPEN.

892
00:43:30,064 --> 00:43:31,766
50% OF THE STUFF YOU GET OUT IS 

893
00:43:31,766 --> 00:43:34,268
INITIATED IN THE WRONG PLACE.

894
00:43:34,268 --> 00:43:38,272
WHAT THIS TELLS US IS ABOUT 

895
00:43:38,272 --> 00:43:39,373
GLOBAL ARCHITECTURE IS IMPORTANT

896
00:43:39,373 --> 00:43:41,509
FOR INITIATION.

897
00:43:41,509 --> 00:43:44,312
SO WHAT REALLY THEN GIVES YOU 

898
00:43:44,312 --> 00:43:45,346
THE PERFECT COMPLEX?

899
00:43:45,346 --> 00:43:49,350
SO HERE IF YOU HAVE TWO RTs AND 

900
00:43:49,350 --> 00:43:52,320
YOU NEED TWO NUCLEIC ACID 

901
00:43:52,320 --> 00:43:53,221
MOLECULES BOUND TO THE LOOPS 

902
00:43:53,221 --> 00:43:57,725
THEY'RE ALSO THE HIGH AFFINITY 

903
00:43:57,725 --> 00:44:00,728
INTERACTIONS NOT THE CHAPERONE 

904
00:44:00,728 --> 00:44:01,762
TAIL INTERACTIONS.

905
00:44:01,762 --> 00:44:03,764
WHEN YOU ASSEMBLE THE COMPLEX 

906
00:44:03,764 --> 00:44:05,166
EVERYTHING YOU GET IS ALMOST 

907
00:44:05,166 --> 00:44:06,267
RACKET.

908
00:44:06,267 --> 00:44:08,536
-- RACKET.

909
00:44:08,536 --> 00:44:09,804
 -- ACCURATE.

910
00:44:09,804 --> 00:44:11,572
WHAT WE STARTED TO GET 

911
00:44:11,572 --> 00:44:12,907
INTERESTED IN IS WHAT DOES THE 

912
00:44:12,907 --> 00:44:14,108
RD MODEL LOOK LIKE?

913
00:44:14,108 --> 00:44:15,810
WHAT WAS FASCINATING WHEN WE DID

914
00:44:15,810 --> 00:44:20,114
THE FIRST TRITRATIONS INTO THE 

915
00:44:20,114 --> 00:44:23,184
NMR IT HAS A COMBINING THAT IS A

916
00:44:23,184 --> 00:44:29,423
TINY AMOUNT OF RT WE EXPECTED 

917
00:44:29,423 --> 00:44:36,364
THE BOTTOM TIER AND DOUBLE 

918
00:44:36,364 --> 00:44:38,599
STRANDED ASSUMED IT WOULD BE 

919
00:44:38,599 --> 00:44:40,301
INTERACTING BUT THAT DOESN'T 

920
00:44:40,301 --> 00:44:40,601
HAPPEN.

921
00:44:40,601 --> 00:44:42,803
THE 18 BASE PAIRS DO NOT 

922
00:44:42,803 --> 00:44:50,311
INTERACT BUT THE U5 STEM THE 

923
00:44:50,311 --> 00:44:51,979
MOTIF ALL SHOW MASSIVE 

924
00:44:51,979 --> 00:44:53,514
PERTURBATIONS.

925
00:44:53,514 --> 00:44:54,315
WE KNEW IT WOULD BIND 

926
00:44:54,315 --> 00:44:55,049
DIFFERENTLY THAN THE OTHER 

927
00:44:55,049 --> 00:44:58,286
STRUCTURES THAT ARE OUT THERE.

928
00:44:58,286 --> 00:45:00,655
SO HERE IS THE DATA THEN THAT 

929
00:45:00,655 --> 00:45:05,159
MELANIE COLLECTED.

930
00:45:05,159 --> 00:45:08,162
AND WE HAVE SOME INFORMATION NOT

931
00:45:08,162 --> 00:45:10,298
COMPLETE BUT  IT ALLOWS US TO 

932
00:45:10,298 --> 00:45:15,736
SEE WHAT IS HAPPENING HERE SO 

933
00:45:15,736 --> 00:45:18,039
THIS IS KNOWN TO HAVE TWO 

934
00:45:18,039 --> 00:45:18,272
STATES.

935
00:45:18,272 --> 00:45:19,440
ONE IS THE BOUND STATE IN WHICH 

936
00:45:19,440 --> 00:45:22,343
THE FINGER AND THUMB ARE 

937
00:45:22,343 --> 00:45:25,279
SEPARATED AND THEN WE KNOW OF 

938
00:45:25,279 --> 00:45:27,315
THE APOS STATE WHERE THE FINGER 

939
00:45:27,315 --> 00:45:28,916
AND THUMB ARE IN CLOSE CONTACT 

940
00:45:28,916 --> 00:45:31,919
AND WHEN THE FINGER AND THUMBS 

941
00:45:31,919 --> 00:45:36,090
ARE OPEN THE ENZYMATIC CLIMBING 

942
00:45:36,090 --> 00:45:37,992
IS AVAILABLE AND NUCLEIC ACIDS 

943
00:45:37,992 --> 00:45:40,328
CAN BIND IN AND YOU CAN SEE WITH

944
00:45:40,328 --> 00:45:44,398
THE MOVEMENT YOU GET APO AND 

945
00:45:44,398 --> 00:45:44,699
BOUND STATE.

946
00:45:44,699 --> 00:45:46,701
IN THE MIDDLE IS A STRUCTURE FOR

947
00:45:46,701 --> 00:45:47,969
THE REMODELER.

948
00:45:47,969 --> 00:45:52,607
WHAT YOU'LL SEE HERE AGAIN IN 

949
00:45:52,607 --> 00:45:54,308
THE LEFT AND RIGHT ALL 

950
00:45:54,308 --> 00:45:58,279
STRUCTURES WE KNOW OF RIGHT NOW 

951
00:45:58,279 --> 00:46:01,616
ARE WHERE THE NUCLEIC ACID BINDS

952
00:46:01,616 --> 00:46:03,751
IN THE DEEP BINDING TARGET BUT 

953
00:46:03,751 --> 00:46:06,020
IF YOU LOOK AT THE MODELER IT'S 

954
00:46:06,020 --> 00:46:07,788
ACTUALLY BINDING IN A COMPLETELY

955
00:46:07,788 --> 00:46:09,757
DIFFERENT WAY.

956
00:46:09,757 --> 00:46:12,727
IT'S ACTUALLY IN THE CLOSE STATE

957
00:46:12,727 --> 00:46:14,929
AND THERE IS A BINDING 

958
00:46:14,929 --> 00:46:16,764
INTERACTION THAT HAPPENS MOSTLY 

959
00:46:16,764 --> 00:46:21,769
BY THE THUMB DOMAIN AND I'LL 

960
00:46:21,769 --> 00:46:26,274
SHOW YOU A LITTLE BIT OF THAT.

961
00:46:26,274 --> 00:46:27,942
THERE'S A DIFFERENT WHEEL AND 

962
00:46:27,942 --> 00:46:36,517
YOU GET THE ENZYMAIYMATIC CLASPY

963
00:46:36,517 --> 00:46:38,319
FROM THE BINDING SITE AND THE 

964
00:46:38,319 --> 00:46:48,729
HELIX IS SITTING THERE.

965
00:47:02,310 --> 00:47:05,179
WHAT WE THEN DID IS TOOK THE 

966
00:47:05,179 --> 00:47:08,349
THUMB AND SAID CAN WE JUST 

967
00:47:08,349 --> 00:47:09,350
LOOK -- THE RESOLUTION WASN'T 

968
00:47:09,350 --> 00:47:13,788
THAT GREAT WE WANTED TO SEE IF 

969
00:47:13,788 --> 00:47:16,324
YOU CAN GET MORE DETAILS BY 

970
00:47:16,324 --> 00:47:21,762
LOOKING AT NMR AND WITH THE 

971
00:47:21,762 --> 00:47:26,267
THUMB DOMAIN WE SAW MANY 

972
00:47:26,267 --> 00:47:27,368
BEAUTIFUL STRUCTURES AND I'LL 

973
00:47:27,368 --> 00:47:32,506
SHOW YOU WHAT IT LOOKS LIKE.

974
00:47:32,506 --> 00:47:38,012
IN THE BLUE IS BOUND.

975
00:47:38,012 --> 00:47:41,315
IN THE GREEN IS HOW THE THUMB 

976
00:47:41,315 --> 00:47:46,253
WOULD BE IN ITS NATURAL FORM AND

977
00:47:46,253 --> 00:47:48,189
THIS LOOP BETWEEN H AND THUMB 

978
00:47:48,189 --> 00:47:49,623
OUT OF THE RESTING PLACE AND YOU

979
00:47:49,623 --> 00:48:00,167
GET THE ARGININES AND LYSINE AND

980
00:48:09,844 --> 00:48:20,321
THIS STRUCTURE IS DIFFERENT WE 

981
00:48:26,327 --> 00:48:36,804
MADE THE TWO MUTANTS LYSINE AND 

982
00:48:37,204 --> 00:48:40,341
ARGININE AND YOU GET ONLY ONE RT

983
00:48:40,341 --> 00:48:42,276
BINDING TO THE HELIX 2 STILL AND

984
00:48:42,276 --> 00:48:45,112
THE REMODELLING DOESN'T HAPPEN 

985
00:48:45,112 --> 00:48:47,815
SO THE SECOND COMES IN.

986
00:48:47,815 --> 00:48:51,085
IN THAT SCENARIO YOU GET NOTHING

987
00:48:51,085 --> 00:48:51,886
STARTING CORRECTLY.

988
00:48:51,886 --> 00:48:56,457
IF YOU MAKE THE LYSINE AND 

989
00:48:56,457 --> 00:48:57,358
ARGININE MUTATION YOU GET THE 

990
00:48:57,358 --> 00:48:57,758
SAME THING.

991
00:48:57,758 --> 00:49:02,997
GET SOME BINDING ON BUT THEN THE

992
00:49:02,997 --> 00:49:08,569
RNA'S NOT REMODELLED AND 

993
00:49:08,569 --> 00:49:10,971
INITIATION DOESN'T GO ANYWHERE.

994
00:49:10,971 --> 00:49:12,573
THAT IT DOUBLE MUTANT SEEMS TO 

995
00:49:12,573 --> 00:49:17,211
BE PRETTY BAD FOR INFECTIVITY.

996
00:49:17,211 --> 00:49:19,914
AND WE SAW THE STRUCTURE OF THAT

997
00:49:19,914 --> 00:49:24,285
DOUBLE MUTANT AND THAT ONE NOW 

998
00:49:24,285 --> 00:49:27,288
BINDS LIKE REGULAR RT IT GOES 

999
00:49:27,288 --> 00:49:29,490
DEEP IN THE ENZYMATIC POCKET AND

1000
00:49:29,490 --> 00:49:32,793
TO ME IT LOOKS LIKE THE ENDS ARE

1001
00:49:32,793 --> 00:49:36,430
NOW COMPLETELY CLEARED FROM THE 

1002
00:49:36,430 --> 00:49:39,533
RNA H AND IT WILL BE HAPPY AND 

1003
00:49:39,533 --> 00:49:43,270
THERE WON'T BE REMODELLING AT 

1004
00:49:43,270 --> 00:49:45,906
THE JUNCTION AND THAT EXPLAINS 

1005
00:49:45,906 --> 00:49:49,977
WHY THE SECOND RT DOESN'T COME 

1006
00:49:49,977 --> 00:49:51,378
IN.

1007
00:49:51,378 --> 00:49:53,113
SO, OUR CURRENT MODEL IS ALL 

1008
00:49:53,113 --> 00:49:59,620
BASED ON INTERACTIONS WITH THIS 

1009
00:49:59,620 --> 00:50:02,890
ABULGE AND THE NATURAL STATE 

1010
00:50:02,890 --> 00:50:08,496
INTERACTS WITH A-BULGE.

1011
00:50:08,496 --> 00:50:12,433
WHEN WHAT THE CRYO IS SAYING THE

1012
00:50:12,433 --> 00:50:14,568
A BULGE WILL MAKE INTERACTION 

1013
00:50:14,568 --> 00:50:17,004
WITH THE CONNECTION DOMAIN.

1014
00:50:17,004 --> 00:50:20,741
AT THAT POINT WHAT MY HYPOTHESIS

1015
00:50:20,741 --> 00:50:22,643
IS IT WILL CAUSE A HINDRANCE 

1016
00:50:22,643 --> 00:50:27,181
WITH THE RNA H DOMAIN AND THAT'S

1017
00:50:27,181 --> 00:50:30,985
WHAT WE'RE CURRENTLY TESTING.

1018
00:50:30,985 --> 00:50:34,722
AND THEN THE NMR DATA SHOWS THE 

1019
00:50:34,722 --> 00:50:35,956
A BULGE WILL INTERACT WITH THE 

1020
00:50:35,956 --> 00:50:41,529
THUMB DOMAIN TO GIVE A DIFFERENT

1021
00:50:41,529 --> 00:50:44,765
KIND OF STEP 2 AFTER WHICH THE 

1022
00:50:44,765 --> 00:50:48,502
REMODELLING HAPPENS AND THEN THE

1023
00:50:48,502 --> 00:50:49,370
SECOND RT BINDS.

1024
00:50:49,370 --> 00:50:53,774
AND THEN FINALLY I WANT TO LEAVE

1025
00:50:53,774 --> 00:50:55,976
YOU WITH WHAT HAPPENS ONCE THE 

1026
00:50:55,976 --> 00:50:59,613
WHOLE SYSTEM HAS ASSEMBLED.

1027
00:50:59,613 --> 00:51:01,048
LIKE THE FIRST EXPERIMENT WE DID

1028
00:51:01,048 --> 00:51:05,319
THIS TO SEE WHEN THE FIRST 

1029
00:51:05,319 --> 00:51:08,188
REMODELER WILL BE BUMPED BY THE 

1030
00:51:08,188 --> 00:51:09,790
ENZYMATIC RT COMING OUT AND WE 

1031
00:51:09,790 --> 00:51:11,592
CAN SAY IT GOES ALL THE WAY UP 

1032
00:51:11,592 --> 00:51:15,396
TO THIS GAC STRETCH BEFORE IT IS

1033
00:51:15,396 --> 00:51:20,801
KICKED OUT.

1034
00:51:20,801 --> 00:51:25,139
BUT DURING THAT PROCESS, 

1035
00:51:25,139 --> 00:51:26,574
INTERESTINGLY WHAT HAPPENS IS 

1036
00:51:26,574 --> 00:51:31,645
THE 5 PREM REGION FORMS A MOTIF 

1037
00:51:31,645 --> 00:51:38,285
AND THIS M5 IS NOW IN CONTACT 

1038
00:51:38,285 --> 00:51:42,523
WITH THE THUMB DOMAIN AFTER THE 

1039
00:51:42,523 --> 00:51:44,091
BENDING.

1040
00:51:44,091 --> 00:51:45,793
PRESUMABLY WE THINK M6 IS ALSO 

1041
00:51:45,793 --> 00:51:49,597
GOING TO BE IN CONTACT WITH THE 

1042
00:51:49,597 --> 00:51:49,797
FIRST.

1043
00:51:49,797 --> 00:51:52,933
WHAT THAT SUGGESTS TO ME IS 

1044
00:51:52,933 --> 00:51:54,001
THROUGHOUT THE INITIATION 

1045
00:51:54,001 --> 00:51:56,337
PROCESS THE FIRST RT IS NOT ONLY

1046
00:51:56,337 --> 00:51:57,871
HELPING THE REMODELLING PROCESS 

1047
00:51:57,871 --> 00:52:03,110
BUT ALSO MAKING SURE THAT IT IS 

1048
00:52:03,110 --> 00:52:08,082
ORCHESTRATING ALL THE STRUCTURE 

1049
00:52:08,082 --> 00:52:10,584
FORMATIONS AND INTERACTION.

1050
00:52:10,584 --> 00:52:12,019
FINALLY, THESE THINGS COME WITH 

1051
00:52:12,019 --> 00:52:14,154
A COST.

1052
00:52:14,154 --> 00:52:17,691
FOR EXAMPLE, WHEN YOU FORM THESE

1053
00:52:17,691 --> 00:52:19,960
MOTIFS RT THEN HAS A PROBLEM 

1054
00:52:19,960 --> 00:52:25,099
GOING THROUGH THAT AND YOU CAN 

1055
00:52:25,099 --> 00:52:26,667
SEE THAT WITH THIS DROP HOWEVER 

1056
00:52:26,667 --> 00:52:29,837
IF YOU HAVE THE TWO NCs SITTING 

1057
00:52:29,837 --> 00:52:36,443
ON THE TRNA YOU HAVE THAT.

1058
00:52:36,443 --> 00:52:38,712
AND MADE FOR THE RNA TO GO 

1059
00:52:38,712 --> 00:52:40,948
THROUGH THE WHOLE THING 

1060
00:52:40,948 --> 00:52:45,786
ACCURATELY YOU NEED ALL OF THIS 

1061
00:52:45,786 --> 00:52:46,286
IN THE RIGHT PLACE.

1062
00:52:46,286 --> 00:52:49,289
SO WITH THAT THE MOST IMPORTANT 

1063
00:52:49,289 --> 00:52:51,792
FEATURE I WANT TO HIGHLIGHT IS 

1064
00:52:51,792 --> 00:52:55,963
THAT DISCOVERY OF THIS RT 

1065
00:52:55,963 --> 00:52:57,231
REMODELER FUNCTION WHICH IS 

1066
00:52:57,231 --> 00:52:59,867
DIFFERENT THAN BOTH STRUCTURALLY

1067
00:52:59,867 --> 00:53:00,868
AND FUNCTIONALLY.

1068
00:53:00,868 --> 00:53:02,469
WE THINK WE HIGHLIGHTED A ROLE 

1069
00:53:02,469 --> 00:53:06,040
FOR RT THAT IS NOT AN ENZYME.

1070
00:53:06,040 --> 00:53:08,275
IT'S A REEL STRUCTURAL REMODELER

1071
00:53:08,275 --> 00:53:11,512
AND THE MOST IMPORTANT THING IS 

1072
00:53:11,512 --> 00:53:14,281
WE CAME UP WITH A NEW RNA 

1073
00:53:14,281 --> 00:53:17,084
BINDING SITE WE DIDN'T KNOW 

1074
00:53:17,084 --> 00:53:19,720
EXISTED AND NEW MODE OF 

1075
00:53:19,720 --> 00:53:20,287
INTERACTION.

1076
00:53:20,287 --> 00:53:22,256
MY OPINION IS WE NEED TO STUDY 

1077
00:53:22,256 --> 00:53:26,260
THE ENTIRE INITIATION COMPLEX IN

1078
00:53:26,260 --> 00:53:30,264
ITS ENTIRE COMPLEX TO FIGURE OUT

1079
00:53:30,264 --> 00:53:33,500
HOW THE WHOLE PROCESS GOES.

1080
00:53:33,500 --> 00:53:44,011
WITH THAT I THINK I'M ON TIME.

1081
00:53:46,080 --> 00:53:47,347
[APPLAUSE]

1082
00:53:47,347 --> 00:53:57,758
>> PLEASE COME ON STAGE.

1083
00:54:04,164 --> 00:54:04,231
 

1084
00:54:04,231 --> 00:54:06,366
>> THOROUGHLY AMAZING ALL
OF 

1085
00:54:06,366 --> 00:54:07,201
YOUR TALKS, THANK YOU.

1086
00:54:07,201 --> 00:54:11,772
I HAVE QUESTIONS FOR ALL OF YOU 

1087
00:54:11,772 --> 00:54:13,407
BUT PAUL YOU GAVE A COMPLEX 

1088
00:54:13,407 --> 00:54:15,809
DESCRIPTION OF ZAP AND ACTIVITY 

1089
00:54:15,809 --> 00:54:24,752
AND SPECIFICITY AND YOU PROBABLY

1090
00:54:24,752 --> 00:54:25,452
THANKFULLY ELIMINATED THE ROLE 

1091
00:54:25,452 --> 00:54:28,388
OF UBIQUITIN ON THE BINDING.

1092
00:54:28,388 --> 00:54:30,457
THERE'S MANY E2s.

1093
00:54:30,457 --> 00:54:33,827
HOW WELL DO WE KNOW THAT'S THE 

1094
00:54:33,827 --> 00:54:38,265
ONLY RELEVANT FOR ZAP AND IS 

1095
00:54:38,265 --> 00:54:39,833
THERE ORTHOLOGOUS EXPERIMENT 

1096
00:54:39,833 --> 00:54:41,101
WITH UBIQUITIN SHOWING AS THE 

1097
00:54:41,101 --> 00:54:42,870
NOT PLAYING A ROLE.

1098
00:54:42,870 --> 00:54:44,404
TRIM 25 IS AN E3.

1099
00:54:44,404 --> 00:54:51,411
>> SHORT ANSWER IS NO.

1100
00:54:51,411 --> 00:54:56,083
NO INTERACTIONS WITH UBIQUITIN.

1101
00:54:56,083 --> 00:54:57,084
WE HAVEN'T DONE PROTEOME 

1102
00:54:57,084 --> 00:54:58,118
INHIBITERS AND THAT SORT OF 

1103
00:54:58,118 --> 00:55:05,492
THING.

1104
00:55:05,492 --> 00:55:07,728
>> FOR MIKE, SOME CELLULAR

1105
00:55:07,728 --> 00:55:09,897
PROMOTERS I THINK YOU CALL THEM 

1106
00:55:09,897 --> 00:55:19,706
TWINED HAVE A PROPENSITY TO HAVE

1107
00:55:19,706 --> 00:55:23,377
NEARBY SITES IS IT DUE TO 

1108
00:55:23,377 --> 00:55:23,710
SEQUESTATION.

1109
00:55:23,710 --> 00:55:24,311
IS IT KNOWN?

1110
00:55:24,311 --> 00:55:26,313
>> IT'S NOT KNOWN BUT WHAT
IS 

1111
00:55:26,313 --> 00:55:31,351
KNOWN IS THERE ARE CERTAIN GENES

1112
00:55:31,351 --> 00:55:35,455
IF YOU EXPRESS IN ONE CELL TYPE 

1113
00:55:35,455 --> 00:55:38,192
YOU GET 1G AND ANOTHER 3G.

1114
00:55:38,192 --> 00:55:41,795
SO THAT RAISES QUESTIONS HOW 

1115
00:55:41,795 --> 00:55:42,930
THERE COULD BE RESPONSE.

1116
00:55:42,930 --> 00:55:45,899
THERE'S AN AREA THAT NEEDS TO BE

1117
00:55:45,899 --> 00:55:50,604
STUDY MORE CAREFULLY.

1118
00:55:50,604 --> 00:55:52,506
>> SHORT QUESTION FOR
VICTORIA.

1119
00:55:52,506 --> 00:55:54,241
IT'S GREAT TO SEE YOUR MODEL 

1120
00:55:54,241 --> 00:55:55,342
CONTINUING TO EVOLVE.

1121
00:55:55,342 --> 00:55:58,078
WHAT ABOUT ON THE PLUS STRAND 

1122
00:55:58,078 --> 00:55:59,880
FROM THE PPT?

1123
00:55:59,880 --> 00:56:03,450
DO YOU ENVISION STILL A SINGLE 

1124
00:56:03,450 --> 00:56:05,586
MOLECULE OR THINK THERE'S 

1125
00:56:05,586 --> 00:56:06,587
REMODELLING REQUIRED THERE AS 

1126
00:56:06,587 --> 00:56:06,787
WELL?

1127
00:56:06,787 --> 00:56:09,890
>> I DON'T KNOW HOW TO
ANSWER 

1128
00:56:09,890 --> 00:56:13,994
THAT QUESTION.

1129
00:56:13,994 --> 00:56:15,896
I KNOW PEOPLE THINK THERE'S A 

1130
00:56:15,896 --> 00:56:17,965
STRUCTURE THERE AND IT'S 

1131
00:56:17,965 --> 00:56:18,665
POSSIBLE.

1132
00:56:18,665 --> 00:56:23,170
THESE REMODELERS CAN REMODEL 

1133
00:56:23,170 --> 00:56:24,071
THEM TOO BUT WE HAVEN'T TOUCHED 

1134
00:56:24,071 --> 00:56:24,338
THAT YET.

1135
00:56:24,338 --> 00:56:34,581
>> THANK YOU.

1136
00:56:46,693 --> 00:56:49,096
>> FOR PAUL, IS THERE
PRESSURE 

1137
00:56:49,096 --> 00:56:51,098
FOR GCs.

1138
00:56:51,098 --> 00:56:54,835
>> MOST ARE NOT BUT A
SUBSTRATE 

1139
00:56:54,835 --> 00:56:55,302
ARE.

1140
00:56:55,302 --> 00:57:02,609
CONVERSELY IN INSECTS AND 

1141
00:57:02,609 --> 00:57:04,144
ARTHROPODS AND THOSE VIRUSES ARE

1142
00:57:04,144 --> 00:57:07,714
PREDICTED TO BE ZAP SENSITIVE.

1143
00:57:07,714 --> 00:57:09,783
>> ALSO A QUESTION FOR
PAUL.

1144
00:57:09,783 --> 00:57:13,353
I WAS CURIOUS ABOUT YOUR DATA 

1145
00:57:13,353 --> 00:57:17,457
SHOWING KHNYN MAY BE REDUNDANT 

1146
00:57:17,457 --> 00:57:18,792
IN FUNCTION AND DID MAPPING OF 

1147
00:57:18,792 --> 00:57:21,094
KHNYN BUT ARE THEY TRULY 

1148
00:57:21,094 --> 00:57:22,229
REDUNDANT DO YOU THINK OR 

1149
00:57:22,229 --> 00:57:23,263
POSSIBLE CELL TYPE SPECIFIC OR 

1150
00:57:23,263 --> 00:57:25,065
TAR GET DIFFERENCES?

1151
00:57:25,065 --> 00:57:26,667
>> RIGHT.

1152
00:57:26,667 --> 00:57:29,336
WE HAVEN'T DONE A CELL SURVEY 

1153
00:57:29,336 --> 00:57:31,338
CLEARLY IN THE PARTICULAR CELLS 

1154
00:57:31,338 --> 00:57:34,107
THEY USE THEY'RE SORT OF 

1155
00:57:34,107 --> 00:57:34,374
REDUNDANT.

1156
00:57:34,374 --> 00:57:38,412
IN THE RECONSTITUTION ASSAY IT'S

1157
00:57:38,412 --> 00:57:42,683
CLEAR KHNYN IS A BIT MORE POTENT

1158
00:57:42,683 --> 00:57:44,117
AND WHY WE FOCUSSED ON THAT 

1159
00:57:44,117 --> 00:57:44,718
PARTICULAR ONE.

1160
00:57:44,718 --> 00:57:48,288
WE DON'T REALLY HAVE ANY 

1161
00:57:48,288 --> 00:57:50,057
INFORMATION ABOUT BP1.

1162
00:57:50,057 --> 00:57:52,025
>> THANKS.

1163
00:57:52,025 --> 00:57:55,395
>> I HAVE A QUESTION FOR 

1164
00:57:55,395 --> 00:57:55,896
MICHAEL.

1165
00:57:55,896 --> 00:57:58,098
WE KNOW FROM STUDIES IN 

1166
00:57:58,098 --> 00:58:06,440
HEPATITIS C VIRUS FROM DUKE THAT

1167
00:58:06,440 --> 00:58:10,744
6 MA SUPPORTS FUNCTIONING AND IS

1168
00:58:10,744 --> 00:58:14,815
IT DIFFERENT IN THE 1G AND 3G 

1169
00:58:14,815 --> 00:58:14,982
RNA?

1170
00:58:14,982 --> 00:58:15,349
>> I'M NOT SURE.

1171
00:58:15,349 --> 00:58:19,519
I JUST STARTED LOOKING INTO THAT

1172
00:58:19,519 --> 00:58:20,220
LITERATURE.

1173
00:58:20,220 --> 00:58:22,990
I DON'T KNOW WHAT PERCENTAGE OF 

1174
00:58:22,990 --> 00:58:24,658
THE RNAs ARE METHYLATED.

1175
00:58:24,658 --> 00:58:28,695
IS IT A SMALL FRACTION OR ARE 

1176
00:58:28,695 --> 00:58:30,230
THEY ALL OF THEM METHYLATED?

1177
00:58:30,230 --> 00:58:33,734
I WAS UNDER THE IMPRESSION THESE

1178
00:58:33,734 --> 00:58:34,701
ARE PRETTY SMALL FRACTIONS OF 

1179
00:58:34,701 --> 00:58:39,172
THE RNA THAT ARE METHYLATED.

1180
00:58:39,172 --> 00:58:42,175
SO I WOULD LOVE TO DO THAT 

1181
00:58:42,175 --> 00:58:46,480
EXPERIMENT BUT WE HAVE TO HAVE A

1182
00:58:46,480 --> 00:58:48,648
WAY OF MAKING THOSE SPECIFIC 

1183
00:58:48,648 --> 00:58:49,750
ADENOSINES METHYLATED.

1184
00:58:49,750 --> 00:58:51,184
THERE MAY BE A WAY OF DOING THAT

1185
00:58:51,184 --> 00:58:53,353
WITH TECHNOLOGY THAT VICTORIA 

1186
00:58:53,353 --> 00:58:53,854
HAS DEVELOPED.

1187
00:58:53,854 --> 00:58:54,988
BUT WE HAVEN'T TRIED THAT YET.

1188
00:58:54,988 --> 00:58:57,891
IT'S A GOOD IDEA.

1189
00:58:57,891 --> 00:59:02,262
THANKS.

1190
00:59:02,262 --> 00:59:10,404
>> A QUESTION FOR MIKE.

1191
00:59:10,404 --> 00:59:15,876
I WAS SURPRISED TO LEARN THE 

1192
00:59:15,876 --> 00:59:22,249
MECHANISM IS DETERMINING THE 

1193
00:59:22,249 --> 00:59:25,585
VIRUS IS THIS EFFECT SEEN IN 

1194
00:59:25,585 --> 00:59:27,487
PRIMARY CELLS ALSO OR IS THAT 

1195
00:59:27,487 --> 00:59:27,821
SOMETHING -- 

1196
00:59:27,821 --> 00:59:30,390
>> YES.

1197
00:59:30,390 --> 00:59:31,792
>> GOOD TO KNOW.

1198
00:59:31,792 --> 00:59:33,360
>> THAT IS SOMETHING THAT
ALICE 

1199
00:59:33,360 --> 00:59:37,431
IS LOOKING AT AS WELL.

1200
00:59:37,431 --> 00:59:38,165
>> YEAH.

1201
00:59:38,165 --> 00:59:39,666
WE SHOULD TALK MORE ABOUT THAT 

1202
00:59:39,666 --> 00:59:40,967
AND WE HAVE A PAPER WE'LL SEND 

1203
00:59:40,967 --> 00:59:46,473
TO YOU TO REVIEW SHORTLY.

1204
00:59:46,473 --> 00:59:50,277
>> QUESTION FOR MIKE.

1205
00:59:50,277 --> 00:59:55,782
SO FOR VIRUSES THAT HAVE I 

1206
00:59:55,782 --> 00:59:57,884
PACKAGING BUT LOW TRANSLATION 

1207
00:59:57,884 --> 00:59:59,986
ARE THEY MORE EFFECTIVE VARIANTS

1208
00:59:59,986 --> 01:00:03,957
AT THE END OF THE DAY?

1209
01:00:03,957 --> 01:00:06,860
>> THAT'S WHAT WAS SHOWN.

1210
01:00:06,860 --> 01:00:09,296
SHE DID THE SAME THING THAT 

1211
01:00:09,296 --> 01:00:11,231
ALICE AND SERGEI FIGURED OUT HOW

1212
01:00:11,231 --> 01:00:11,832
TO DO.

1213
01:00:11,832 --> 01:00:14,768
THEY WERE ABLE TO MAKE TRANSFECT

1214
01:00:14,768 --> 01:00:16,703
CELLS WITH A VECTOR THAT ONLY 

1215
01:00:16,703 --> 01:00:18,205
MAKES THE 1G.

1216
01:00:18,205 --> 01:00:20,507
AND SO THOSE VIRUSES THAT ARE 

1217
01:00:20,507 --> 01:00:22,509
MADE ARE NOT HEALTHY.

1218
01:00:22,509 --> 01:00:26,480
THEY HAVE A REPLICATION DEFECT.

1219
01:00:26,480 --> 01:00:28,648
AND IF THEY TRANSFECT WITH A 

1220
01:00:28,648 --> 01:00:31,785
VECTOR THAT ONLY MAKES THE 3G 

1221
01:00:31,785 --> 01:00:39,426
THERE'S ALSO A DEFECT.

1222
01:00:39,426 --> 01:00:40,894
NORMALLY THE WILD TYPE VIRUS 

1223
01:00:40,894 --> 01:00:41,194
MAKES BOTH.

1224
01:00:41,194 --> 01:00:42,596
THERE'S A MIXTURE.

1225
01:00:42,596 --> 01:00:44,498
IT WOULD BE INTERESTING TO KNOW 

1226
01:00:44,498 --> 01:00:46,733
IF THE PROPORTIONS CHANGE DURING

1227
01:00:46,733 --> 01:00:48,301
THE VIRAL REPLICATION CYCLE BUT 

1228
01:00:48,301 --> 01:00:49,703
NOBODY HAS BEEN ABLE TO FIGURE 

1229
01:00:49,703 --> 01:00:51,505
OUT HOW TO DO THAT EXPERIMENT 

1230
01:00:51,505 --> 01:00:51,671
YET.

1231
01:00:51,671 --> 01:00:52,005
>> THANK YOU.

1232
01:00:52,005 --> 01:00:57,644
>> QUESTION FOR PAUL.

1233
01:00:57,644 --> 01:01:02,182
IN YOUR INITIAL EXPERIMENT THE 

1234
01:01:02,182 --> 01:01:05,218
CAGE NYN KNOCKOUT DIDN'T REDUCE 

1235
01:01:05,218 --> 01:01:06,987
VIRUS REPLICATION BUT LATER IN 

1236
01:01:06,987 --> 01:01:07,954
THE EXPERIMENT IT WAS HAVING AN 

1237
01:01:07,954 --> 01:01:08,421
EFFECT.

1238
01:01:08,421 --> 01:01:15,095
I DON'T KNOW IF I MISSED THAT.

1239
01:01:15,095 --> 01:01:19,132
IT SEEMS I REQUIRED SINGLE 

1240
01:01:19,132 --> 01:01:20,767
STRANDED RNA TO WALK WAS THAT 

1241
01:01:20,767 --> 01:01:27,607
DIFFERENT THAN THE INITIAL VIRUS

1242
01:01:27,607 --> 01:01:29,509
YOU WORKED WITH.

1243
01:01:29,509 --> 01:01:32,846
>> IT'S IN A FAMILY OF
NUCLEASES

1244
01:01:32,846 --> 01:01:35,949
AND AT LEAST TWO ARE EXPRESSED 

1245
01:01:35,949 --> 01:01:37,484
IN THE CELLS OF THE INITIAL TO 

1246
01:01:37,484 --> 01:01:38,919
SEE THE EFFECT YOU HAVE TO 

1247
01:01:38,919 --> 01:01:39,586
KNOCKOUT BOTH.

1248
01:01:39,586 --> 01:01:45,025
ONCE YOU KNOCKED OUT BOTH YOU 

1249
01:01:45,025 --> 01:01:48,662
CAN PUT ONE BACK AND ANALYZE THE

1250
01:01:48,662 --> 01:01:53,433
FUNCTION THAT WAY.

1251
01:01:53,433 --> 01:01:56,703
>> THANK YOU.

1252
01:01:56,703 --> 01:02:04,778
>> WHAT IS THE THOUGHT OF
CAP 

1253
01:02:04,778 --> 01:02:06,379
INDEPENDENT TRANSLATION.

1254
01:02:06,379 --> 01:02:10,317
>> IT'S BEEN A
CONTROVERSIAL 

1255
01:02:10,317 --> 01:02:13,620
TOPIC A LONG TIME AND WE HAVEN'T

1256
01:02:13,620 --> 01:02:15,021
LOOKED AT IT.

1257
01:02:15,021 --> 01:02:24,331
HIV USES IT IN GENERAL AND COULD

1258
01:02:24,331 --> 01:02:26,633
USE AN IRIS BUT NOT SOMETHING 

1259
01:02:26,633 --> 01:02:27,934
I'VE LOOKED AT.

1260
01:02:27,934 --> 01:02:32,973
>> FOR TRANSLATION THAT
WAY?

1261
01:02:32,973 --> 01:02:33,240
WHO KNOWS.

1262
01:02:33,240 --> 01:02:43,750
>> NEEDS SOME VIROLOGY,
STEVE.

1263
01:02:52,559 --> 01:02:57,063
>> THANK YOU.

1264
01:02:57,063 --> 01:03:05,005
WE HAVE THE NEXT SESSION FROM 

1265
01:03:05,005 --> 01:03:14,180
DUKE WE HAVE PRIYAMVADA AND 

1266
01:03:14,180 --> 01:03:24,457
MUNIR AND GNANA.

1267
01:03:47,447 --> 01:03:54,387
YOU  

1268
01:04:17,344 --> 01:04:17,844
>> ALL RIGHT.

1269
01:04:17,844 --> 01:04:19,512
IT'S A PLEASURE TO INTRODUCE THE

1270
01:04:19,512 --> 01:04:21,548
DUKE CENTER FOR HIV STRUCTURAL 

1271
01:04:21,548 --> 01:04:21,948
BIOLOGY.

1272
01:04:21,948 --> 01:04:24,184
SO THE DUKE CENTER FOR HIV 

1273
01:04:24,184 --> 01:04:26,653
STRUCTURAL BIOLOGY IS FOCUSSED 

1274
01:04:26,653 --> 01:04:31,391
ON THE HH1 ENVELOPE YOU SEE 

1275
01:04:31,391 --> 01:04:31,691
HERE.

1276
01:04:31,691 --> 01:04:33,927
AND COLORED ACCORDING TO SITES 

1277
01:04:33,927 --> 01:04:36,529
OF VULNERABILITY WHERE 

1278
01:04:36,529 --> 01:04:37,731
ANTIBODIES CAN BIND AND 

1279
01:04:37,731 --> 01:04:38,031
NEUTRALIZE.

1280
01:04:38,031 --> 01:04:39,432
WE HAVE THREE PROJECT.

1281
01:04:39,432 --> 01:04:42,102
THE FIRST LOOKS AT THE ROLE OF 

1282
01:04:42,102 --> 01:04:45,038
THE ENVELOPE IN HIV-1 ENTRY.

1283
01:04:45,038 --> 01:04:46,973
THE SECOND LOOK AT HIV-1 

1284
01:04:46,973 --> 01:04:50,276
ENVELOPE INTERACTION WITH THE B 

1285
01:04:50,276 --> 01:04:53,313
CELL RECEPTOR TRIGGERING ANTIGEN

1286
01:04:53,313 --> 01:04:54,714
PRODUCTION AND THE THIRD LOOK AT

1287
01:04:54,714 --> 01:05:02,255
THE ENVELOPE IN CONTROLLING 

1288
01:05:02,255 --> 01:05:03,823
HIV-1 REBOUND FROM THE LATENT 

1289
01:05:03,823 --> 01:05:04,724
RESERVOIR.

1290
01:05:04,724 --> 01:05:06,893
SO THE THREE PROJECTS ARE 

1291
01:05:06,893 --> 01:05:10,063
SUPPORTED BY THREE CORE 

1292
01:05:10,063 --> 01:05:12,699
STRUCTURAL COMPUTATIONAL AND 

1293
01:05:12,699 --> 01:05:13,400
IMMUNOBIOLOGY.

1294
01:05:13,400 --> 01:05:16,403
THE CENTER IS SUPPORTED BY AN 

1295
01:05:16,403 --> 01:05:18,571
ADMINISTRATIVE CORE AND RUNS A 

1296
01:05:18,571 --> 01:05:19,239
DEVELOPMENTAL CORE.

1297
01:05:19,239 --> 01:05:21,107
THAT'S THE STRUCTURE OF THE 

1298
01:05:21,107 --> 01:05:21,608
CENTER.

1299
01:05:21,608 --> 01:05:25,078
THE TWO TALKS YOU'LL HEAR TODAY 

1300
01:05:25,078 --> 01:05:32,318
ARE BY DR. ALAM AND GNANAKARAN 

1301
01:05:32,318 --> 01:05:34,020
AND THE B CELL RECEPTOR PROJECT 

1302
01:05:34,020 --> 01:05:37,957
2 AND SEE IT'S CLOSE WITH 

1303
01:05:37,957 --> 01:05:40,427
STRUCTURAL AND IMMUNOBIOLOGY IN 

1304
01:05:40,427 --> 01:05:40,794
THE TALKS.

1305
01:05:40,794 --> 01:05:43,863
TO GIVE AN IDEA OF WHAT WE ARE 

1306
01:05:43,863 --> 01:05:45,331
DOING AND BIG PICTURE WE'RE 

1307
01:05:45,331 --> 01:05:48,334
TRYING TO UNDERSTAND THE COMPLEX

1308
01:05:48,334 --> 01:05:53,406
PROCESS AND WE'RE TRYING TO 

1309
01:05:53,406 --> 01:05:59,412
UNDERSTAND HOW THE BCR SENDS 

1310
01:05:59,412 --> 01:06:02,882
SIGNAL OVER INCLUDING TO T CELL 

1311
01:06:02,882 --> 01:06:03,249
HELP.

1312
01:06:03,249 --> 01:06:04,951
SO SIGNAL ONE IS THE PART YOU'LL

1313
01:06:04,951 --> 01:06:07,654
HEAR ABOUT TODAY.

1314
01:06:07,654 --> 01:06:10,757
THE SIGNAL 2 ASPECT IS IN 

1315
01:06:10,757 --> 01:06:11,458
COLLABORATION WITH THE 

1316
01:06:11,458 --> 01:06:14,294
UNIVERSITY OF OXFORD.

1317
01:06:14,294 --> 01:06:16,029
YOU'LL PROBABLY HEAR ABOUT IT AT

1318
01:06:16,029 --> 01:06:17,130
A LATER MEETING.

1319
01:06:17,130 --> 01:06:20,600
THERE'S ONLY SO MUCH WE CAN 

1320
01:06:20,600 --> 01:06:22,001
SQUEEZE INTO ONE.

1321
01:06:22,001 --> 01:06:24,337
SO THERE'S THREE STRUCTURES OF 

1322
01:06:24,337 --> 01:06:28,541
PCRs KNOWN SO FAR AND CRYO EM 

1323
01:06:28,541 --> 01:06:29,409
STRUCTURES AND THEN LEAVE A LOT 

1324
01:06:29,409 --> 01:06:30,743
OF QUESTIONS UNANSWERED.

1325
01:06:30,743 --> 01:06:32,879
HOW DOES THE DYNAMICS LOOK 

1326
01:06:32,879 --> 01:06:35,014
BECAUSE YOU HAVE THREE 

1327
01:06:35,014 --> 01:06:38,785
STRUCTURES, TWO IGM AND ONE IGG 

1328
01:06:38,785 --> 01:06:39,085
STRUCTURE.

1329
01:06:39,085 --> 01:06:40,186
THERE ISN'T MUCH TO COMPARE 

1330
01:06:40,186 --> 01:06:40,386
WITH.

1331
01:06:40,386 --> 01:06:43,756
HOW DO THE DYNAMICS LOOK AND HOW

1332
01:06:43,756 --> 01:06:47,794
DOES IT SENSE THE ANTIGEN 

1333
01:06:47,794 --> 01:06:49,829
BINDING AND THEN SENDS THOSE 

1334
01:06:49,829 --> 01:06:52,031
CONFIRMATIONAL SIGNALS TO WHERE 

1335
01:06:52,031 --> 01:06:52,932
IT NEEDS TO SIGNAL.

1336
01:06:52,932 --> 01:06:56,703
THOSE ARE THE GAPS IN KNOWLEDGE 

1337
01:06:56,703 --> 01:06:58,271
WE ARE PLANNING TO FILL WITH 

1338
01:06:58,271 --> 01:07:02,876
PROJECT 2 OVER THE LONG TERM.

1339
01:07:02,876 --> 01:07:08,414
THE PROJECT 2 IS BEING 

1340
01:07:08,414 --> 01:07:16,823
SPEARHEADED BY MONET RALAM AND 

1341
01:07:16,823 --> 01:07:18,892
THEY HAD POSTERS YESTERDAY 

1342
01:07:18,892 --> 01:07:20,026
THEY'RE HERE IF YOU HAVE NOT HAD

1343
01:07:20,026 --> 01:07:22,228
A CHANCE TO TALK TO THEM, PLEASE

1344
01:07:22,228 --> 01:07:22,495
DO.

1345
01:07:22,495 --> 01:07:24,998
THE COMPUTATIONAL ASPECT YOU'LL 

1346
01:07:24,998 --> 01:07:33,039
HEAR ABOUT FROM GNANA HAS BEEN 

1347
01:07:33,039 --> 01:07:43,583
LED AND MING XIO AND BETWEEN KEN

1348
01:07:43,850 --> 01:07:46,052
HIS POSTER WILL BE WELL TAKEN 

1349
01:07:46,052 --> 01:07:52,592
CARE OF AND TALK TWO FROM THE 

1350
01:07:52,592 --> 01:07:54,127
STRUCTURAL BIOLOGY CORE AND 

1351
01:07:54,127 --> 01:07:57,463
ALBERTO BARTESAGHI WILL TALK 

1352
01:07:57,463 --> 01:08:00,500
ABOUT WHAT THEY CALL THE MAGICAL

1353
01:08:00,500 --> 01:08:01,167
NEXT STEP.

1354
01:08:01,167 --> 01:08:02,902
HE'LL TALK ABOUT HIS 

1355
01:08:02,902 --> 01:08:06,139
COMPUTATIONAL PIPELINE FOR 

1356
01:08:06,139 --> 01:08:08,942
PROCESSING CRYO ET DATA.

1357
01:08:08,942 --> 01:08:11,678
SUPPORTING ALBERTO IN THIS 

1358
01:08:11,678 --> 01:08:12,879
EFFORT ARE TWO GRADUATE STUDENTS

1359
01:08:12,879 --> 01:08:14,781
FROM HIS LAB LAURA AND ABBY.

1360
01:08:14,781 --> 01:08:16,482
THEY HAVE POSTERS TODAY SO 

1361
01:08:16,482 --> 01:08:18,885
PLEASE STOP BY AND LEARN MORE 

1362
01:08:18,885 --> 01:08:20,353
ABOUT THE MAGICAL NEXT STEP.

1363
01:08:20,353 --> 01:08:21,955
SO NOW THESE ARE THE TALKS 

1364
01:08:21,955 --> 01:08:22,522
YOU'LL HEAR ABOUT TODAY.

1365
01:08:22,522 --> 01:08:24,958
I ALSO WANTED TO GIVE YOU A 

1366
01:08:24,958 --> 01:08:27,293
GLIMPSE OF WHAT ELSE IS HOT AND 

1367
01:08:27,293 --> 01:08:27,994
HAPPENING IN THE CENTER BEYOND 

1368
01:08:27,994 --> 01:08:31,230
THESE TALKS WE HAVE TIME TO 

1369
01:08:31,230 --> 01:08:36,703
INCORPORATE IN TODAY'S MEETING.

1370
01:08:36,703 --> 01:08:38,237
PROJECT 1 IS CLOSELY SYNERGIZED 

1371
01:08:38,237 --> 01:08:39,305
WITH THE CORE AND WE'RE LOOKING 

1372
01:08:39,305 --> 01:08:41,874
AT MANY THINGS BUT THE CRUX OF 

1373
01:08:41,874 --> 01:08:43,476
THE PROJECT IS WILL TIME SCALE 

1374
01:08:43,476 --> 01:08:43,843
DYNAMICS.

1375
01:08:43,843 --> 01:08:46,079
WE'RE LOOKING AT THE PROCESSING 

1376
01:08:46,079 --> 01:08:49,248
IN HIV-1 ENTRY WHICH HAPPEN TOO 

1377
01:08:49,248 --> 01:08:50,283
FA OF THE FOR CONVENTIONAL 

1378
01:08:50,283 --> 01:08:52,051
METHODS TO CAPTURE THEM.

1379
01:08:52,051 --> 01:08:57,256
TWO LABS ARE LEADING AT THE 

1380
01:08:57,256 --> 01:08:58,625
FOCUS OF THIS EFFORT AND ONE IS 

1381
01:08:58,625 --> 01:09:01,127
FROM SWITZERLAND.

1382
01:09:01,127 --> 01:09:05,365
YOU SEE HERE HIM ON THE LEFT 

1383
01:09:05,365 --> 01:09:06,833
WITH TWO STUDENTS AND THEN THE 

1384
01:09:06,833 --> 01:09:09,002
LAB OF RORY HENDERSON FROM DUKE 

1385
01:09:09,002 --> 01:09:10,737
UNIVERSITY AND SEE HIS LAB ON 

1386
01:09:10,737 --> 01:09:13,573
THE RIGHT WITH THEIR LAB 

1387
01:09:13,573 --> 01:09:13,906
MEMBERS.

1388
01:09:13,906 --> 01:09:15,842
AND THEY'RE DOING TEMPERATURE 

1389
01:09:15,842 --> 01:09:18,611
JUMP CRYO EM AND AT DUKE RORY IS

1390
01:09:18,611 --> 01:09:21,914
DOING JUMPS ACROSS THE AXIS AND 

1391
01:09:21,914 --> 01:09:24,484
SIMULATION TO CAPTURE THESE FAST

1392
01:09:24,484 --> 01:09:26,285
INTERMEDIATES.

1393
01:09:26,285 --> 01:09:28,254
I WANTED TO GIVE A QUICK 

1394
01:09:28,254 --> 01:09:30,957
OVERVIEW OF THE CRYO EM AND I'LL

1395
01:09:30,957 --> 01:09:31,824
TELL YOU WHY.

1396
01:09:31,824 --> 01:09:33,893
FOR AN OVERVIEW YOU FREEZE THE 

1397
01:09:33,893 --> 01:09:38,197
SAMPLE ON THE GRID AND EXCITE IT

1398
01:09:38,197 --> 01:09:40,066
BY PULSING A LASER SO THE ICE 

1399
01:09:40,066 --> 01:09:42,802
AROUND IT MELTS AND YOU 

1400
01:09:42,802 --> 01:09:44,804
STIMULATE IT WITH A SMALL 

1401
01:09:44,804 --> 01:09:45,705
MOLECULE OR WHATEVER.

1402
01:09:45,705 --> 01:09:47,640
THE MOLECULE MOVES AND BEFORE IT

1403
01:09:47,640 --> 01:09:49,942
HAS A CHANCE TO FIGURE OUT WHAT 

1404
01:09:49,942 --> 01:09:52,078
JUST HAPPENED YOU FREEZE IT BACK

1405
01:09:52,078 --> 01:09:54,914
AND THEN HOPEFULLY CAPTURE AN 

1406
01:09:54,914 --> 01:09:55,248
INTERMEDIATE.

1407
01:09:55,248 --> 01:09:57,316
THAT'S THE CRUX THE TECHNOLOGY.

1408
01:09:57,316 --> 01:09:59,385
THEY'VE BEEN ABLE TO ESTABLISH 

1409
01:09:59,385 --> 01:10:02,255
THE TECHNOLOGY IN MANY SYSTEMS 

1410
01:10:02,255 --> 01:10:03,389
FOR ENVELOPE WE'RE STILL TRYING 

1411
01:10:03,389 --> 01:10:05,525
TO FIGURING OUT SOME OF THE 

1412
01:10:05,525 --> 01:10:10,363
THINGS LIKE HOW TO MINIMIZE 

1413
01:10:10,363 --> 01:10:12,632
REPARATION AND THESE ARE 

1414
01:10:12,632 --> 01:10:14,133
ENVELOPE SPECIFIC THINGS AND 

1415
01:10:14,133 --> 01:10:18,104
LEAD TO TECHNOLOGY DEVELOPMENT 

1416
01:10:18,104 --> 01:10:18,404
APPLICABLE.

1417
01:10:18,404 --> 01:10:21,207
BUT FOR NOW THERE IS THE 

1418
01:10:21,207 --> 01:10:23,342
TECHNOLOGY THERE.

1419
01:10:23,342 --> 01:10:29,449
IT RICK LAURENS STUDIED THE 

1420
01:10:29,449 --> 01:10:30,850
CAPSID SYSTEM AND I HEARD THE 

1421
01:10:30,850 --> 01:10:32,752
TALKS GOING ON THE CAPSID HERE 

1422
01:10:32,752 --> 01:10:34,087
IN THE MEETING.

1423
01:10:34,087 --> 01:10:35,988
I FELT LIKE A LOT OF THE STUDIES

1424
01:10:35,988 --> 01:10:37,790
THAT I HEARD WOULD BENEFIT FROM 

1425
01:10:37,790 --> 01:10:38,291
THIS TECHNOLOGY.

1426
01:10:38,291 --> 01:10:39,759
SO I'LL PUT IT OUT THERE FOR YOU

1427
01:10:39,759 --> 01:10:41,461
ALL.

1428
01:10:41,461 --> 01:10:47,300
SO THIS IS BASICALLY FROM HIS 

1429
01:10:47,300 --> 01:10:49,335
REVIEW RECENTLY WHICH TELLS YOU 

1430
01:10:49,335 --> 01:10:51,738
WHAT THE TECHNIQUE CAN DO AND 

1431
01:10:51,738 --> 01:10:53,172
CAPTURE INTERMEDIATES YOU WOULD 

1432
01:10:53,172 --> 01:10:55,708
NOT SEE OTHERWISE.

1433
01:10:55,708 --> 01:10:56,876
THAT WAS THE FOCUS OF 

1434
01:10:56,876 --> 01:10:57,343
HIGHLIGHTING THIS.

1435
01:10:57,343 --> 01:11:00,446
WE ARE WORKING ON IT AND 

1436
01:11:00,446 --> 01:11:03,282
INVESTING TIME ON MAKING THE 

1437
01:11:03,282 --> 01:11:06,119
METHOD TO ENVELOPE.

1438
01:11:06,119 --> 01:11:12,425
IT CAN BE USED FOR OTHER THINGS.

1439
01:11:12,425 --> 01:11:15,928
AND YOU CAN SEE THE STRUCTURAL 

1440
01:11:15,928 --> 01:11:25,438
AND IMMUNOBIOLOGY CORE THE 

1441
01:11:25,438 --> 01:11:28,007
PHILOSOPHY IS THROUGH THIS.

1442
01:11:28,007 --> 01:11:30,276
THE SCIENTIFIC QUESTION PROJECT 

1443
01:11:30,276 --> 01:11:35,915
3 ASKING IS THE ROLE OF 

1444
01:11:35,915 --> 01:11:37,984
ORTHOLOGOUS ANTIBODY REBOUND.

1445
01:11:37,984 --> 01:11:40,853
IT'S CLOSELY ENMESHED WITH 

1446
01:11:40,853 --> 01:11:42,288
MULTIPLE INSTITUTIONS AND CORES.

1447
01:11:42,288 --> 01:11:44,957
HOW IS THIS BEING DONE?

1448
01:11:44,957 --> 01:11:47,393
THE SAMPLES AND THE BIOLOGY ARE 

1449
01:11:47,393 --> 01:11:52,098
ALL COMING FROM THE CURE WORLD 

1450
01:11:52,098 --> 01:11:55,701
ON ORTHOLOGOUS ANTIBODIES AND 

1451
01:11:55,701 --> 01:11:58,271
THE ISOLATION TECHNOLOGIES WHICH

1452
01:11:58,271 --> 01:11:59,539
HAVE BEEN WELL DEVELOPED IN THE 

1453
01:11:59,539 --> 01:12:02,542
VACCINE WORLD ARE HELPING US 

1454
01:12:02,542 --> 01:12:04,410
ISOLATE THESE ANTIBODIES AND 

1455
01:12:04,410 --> 01:12:07,013
STUDY THEM.

1456
01:12:07,013 --> 01:12:10,116
THE EPITOPE MAPPING AND THE 

1457
01:12:10,116 --> 01:12:10,850
STRUCTURAL BIOLOGY TECHNIQUES 

1458
01:12:10,850 --> 01:12:17,156
WELL ESTABLISHED IN THIS 

1459
01:12:17,156 --> 01:12:19,292
RELEVANT QUESTION OF HOW 

1460
01:12:19,292 --> 01:12:19,892
ORTHOLOGOUS ANTIBODIES BLOCK 

1461
01:12:19,892 --> 01:12:23,863
REBOUND AND WHAT CAN WE DO TO 

1462
01:12:23,863 --> 01:12:26,532
BLOCK THE REBOUND OF THE VIRUSES

1463
01:12:26,532 --> 01:12:31,470
NOT BLOCKED BY THE ORTHOLOGOUS 

1464
01:12:31,470 --> 01:12:31,838
ANTIBODIES?

1465
01:12:31,838 --> 01:12:34,874
SO THEN OUR PROJECT 3 TEAM IS 

1466
01:12:34,874 --> 01:12:35,074
HERE.

1467
01:12:35,074 --> 01:12:37,043
THIS IS THE PANEL LAB MEMBERS 

1468
01:12:37,043 --> 01:12:37,977
HERE YESTERDAY AND UNFORTUNATELY

1469
01:12:37,977 --> 01:12:41,113
I DON'T THINK THEY'RE HERE TODAY

1470
01:12:41,113 --> 01:12:43,316
BUT ON ZOOM OR STREAM AND THEY 

1471
01:12:43,316 --> 01:12:45,484
HAVE POSTERS AND KAYLA WILSON 

1472
01:12:45,484 --> 01:12:48,287
WILL COVER THE IMMUNOBIOLOGY 

1473
01:12:48,287 --> 01:12:51,023
ASPECT OF THE WORK FROM MY LAB 

1474
01:12:51,023 --> 01:12:56,329
ELI ZANG WILL PRESENT A 

1475
01:12:56,329 --> 01:13:06,839
STRUCTURE OF AN ENVELOPE AND 

1476
01:13:08,975 --> 01:13:11,177
THIS WAS A NICE PICTURE SO I PUT

1477
01:13:11,177 --> 01:13:15,448
IT THERE AND TWO ARE POSTERS.

1478
01:13:15,448 --> 01:13:18,284
MARLENE AND LOUISA AND YOU SEE 

1479
01:13:18,284 --> 01:13:20,519
THEM BONDING AT OUR RECENT 

1480
01:13:20,519 --> 01:13:25,758
ANNUAL MEETING AND WITH THAT I 

1481
01:13:25,758 --> 01:13:30,296
WILL ASK FOR HIM TO COME DO HIS 

1482
01:13:30,296 --> 01:13:40,406
TALK.

1483
01:13:47,246 --> 01:13:51,851
>> SO I NEED TO TALK ABOUT
SOME 

1484
01:13:51,851 --> 01:13:56,989
OF OUR RECENT WORKS WHERE WE ARE

1485
01:13:56,989 --> 01:13:59,091
LOOKING AT THE STRUCTURE AND 

1486
01:13:59,091 --> 01:14:04,764
SIGNALLING OF B CELL RECEPTORS 

1487
01:14:04,764 --> 01:14:08,534
EXPRESSING BROADLY NEUTRALIZING 

1488
01:14:08,534 --> 01:14:09,168
ANTIBODY.

1489
01:14:09,168 --> 01:14:19,545
IT'S A HETTERO METRIC 

1490
01:14:20,813 --> 01:14:22,982
DIMERIZATION AND IT'S SYMMETRIC 

1491
01:14:22,982 --> 01:14:24,583
BETWEEN THE IMMUNOGLOBULIN AND 

1492
01:14:24,583 --> 01:14:28,054
THE BETA AND THIS FUNCTIONS AS A

1493
01:14:28,054 --> 01:14:30,056
SIGNALLING COMPONENT OF THE BCR 

1494
01:14:30,056 --> 01:14:35,895
COMPLEX THROUGH THE I10 MOTIF IN

1495
01:14:35,895 --> 01:14:44,036
THE ISOLIC TABLE AND ADOPTS THE 

1496
01:14:44,036 --> 01:14:45,838
ARCHITECTURE OF THE SOLUBLE 

1497
01:14:45,838 --> 01:14:46,439
ANTIBODY.

1498
01:14:46,439 --> 01:14:50,276
WHEN WE THINK WILL ANTIBODY 

1499
01:14:50,276 --> 01:14:52,311
SHAPE THE CANONICAL Y SHAPE IS 

1500
01:14:52,311 --> 01:14:58,751
WHAT WE'RE FAMILIAR WITH AND 

1501
01:14:58,751 --> 01:15:02,021
THEY ADOPT A CONFIGURATION TO 

1502
01:15:02,021 --> 01:15:04,824
BIND TO CLOSE THE SPACE GLYCANS 

1503
01:15:04,824 --> 01:15:14,567
AND WITH STRONGER AFFINITY.

1504
01:15:14,567 --> 01:15:25,111
AROUND THE 2G AND HAVE SEEN THE 

1505
01:15:26,345 --> 01:15:29,048
NON-DOMAIN SWAP FDG ANTIBODIES 

1506
01:15:29,048 --> 01:15:31,784
AROUND ARE CURRENTLY WORKING TO 

1507
01:15:31,784 --> 01:15:35,388
GET BCR STRUCTURES FOR THEM AS 

1508
01:15:35,388 --> 01:15:35,588
WELL.

1509
01:15:35,588 --> 01:15:38,858
WE HAVE EARLIER HYPOTHESIZED THE

1510
01:15:38,858 --> 01:15:43,996
BCRs OF GLYCAN BINDING 

1511
01:15:43,996 --> 01:15:46,599
ANTIBODIES ADOPT CONFIGURATIONS 

1512
01:15:46,599 --> 01:15:50,403
AND HAVE STRUCTURALLY DISTINCT 

1513
01:15:50,403 --> 01:15:53,539
WILL FORMATIONS AND THE 

1514
01:15:53,539 --> 01:15:57,276
SIGNALLING WOULD BE DISTINCT.

1515
01:15:57,276 --> 01:16:00,880
WE EXPRESSED B CELL RECEPTORS ON

1516
01:16:00,880 --> 01:16:08,921
B CELLS AND THESE WERE AFFINITY 

1517
01:16:08,921 --> 01:16:12,658
PURIFIED AND BY PHYSICAL 

1518
01:16:12,658 --> 01:16:15,561
ANALYSIS INDICATED THE PURIFIED 

1519
01:16:15,561 --> 01:16:18,931
BCRs IN MYCELLAR FORMS IS INTACT

1520
01:16:18,931 --> 01:16:24,437
WITH THE BETA COMPONENT AND THE 

1521
01:16:24,437 --> 01:16:34,914
ANALYSIS SHOWED THE SOLUBLE 

1522
01:16:37,716 --> 01:16:39,351
ANTIBODIES AND AS EXPECTED IT 

1523
01:16:39,351 --> 01:16:44,256
ADOPTS THE CANONICAL Y STHAP.

1524
01:16:44,256 --> 01:16:44,323
 

1525
01:16:46,859 --> 01:16:47,793
-- SHAPE.

1526
01:16:47,793 --> 01:16:54,500
AND WANTED TO MAKE SURE IT BINDS

1527
01:16:54,500 --> 01:17:05,010
TO THE RELEVANT AND THE BOUND 

1528
01:17:05,444 --> 01:17:07,213
THE ANTIBODY WITH DIFFERENT 

1529
01:17:07,213 --> 01:17:10,616
AFFINITIES AND THE IGG AND IGM 

1530
01:17:10,616 --> 01:17:16,088
PCRs BIND WITH THE SAME 

1531
01:17:16,088 --> 01:17:16,589
AFFINITY.

1532
01:17:16,589 --> 01:17:19,325
THE COMPLEX IS PURIFIED AND 

1533
01:17:19,325 --> 01:17:20,960
FUNCTIONAL IN RECOGNIZING 

1534
01:17:20,960 --> 01:17:23,863
RELEVANT HIV TRIMERS.

1535
01:17:23,863 --> 01:17:29,101
IN THE NEXT SLIDES I'LL GO OVER 

1536
01:17:29,101 --> 01:17:35,007
WHAT WE KNOW FROM THE TWO BRR 

1537
01:17:35,007 --> 01:17:36,242
STRUCTURES RECENTLY REPORTED AND

1538
01:17:36,242 --> 01:17:40,079
WHAT WE DO NOT KNOW.

1539
01:17:40,079 --> 01:17:44,116
SO THIS IS A MOUSE BGM FOR A 

1540
01:17:44,116 --> 01:17:48,053
MOLECULE AND WHAT WE LEARNED 

1541
01:17:48,053 --> 01:17:54,760
FROM THIS STRUCTURE IS THAT 

1542
01:17:54,760 --> 01:17:59,465
FIRST THE ASYMMETRIC 

1543
01:17:59,465 --> 01:18:02,268
STOICHIOMETRY DEFINED BY 

1544
01:18:02,268 --> 01:18:04,537
CHEMICAL STUDIES WAS CONFORMED 

1545
01:18:04,537 --> 01:18:13,612
IN THE BCR STRUCTURE.

1546
01:18:13,612 --> 01:18:15,748
WE LEARNED AND THE INFORMATION 

1547
01:18:15,748 --> 01:18:19,151
CAME LARGELY FROM THE TRUNCATED 

1548
01:18:19,151 --> 01:18:20,853
BCR DUE TO THE FLEXIBILITY OF 

1549
01:18:20,853 --> 01:18:24,857
THE FAB AND THAT WAS REPORTED 

1550
01:18:24,857 --> 01:18:29,161
HOWEVER, THE BCR DYNAMICS 

1551
01:18:29,161 --> 01:18:31,497
INFORMATION WAS NOT STUDIED DUE 

1552
01:18:31,497 --> 01:18:35,968
TO TECHNICAL HURDLES AND THUS WE

1553
01:18:35,968 --> 01:18:38,270
HAVE NO INFORMATION ABOUT THE 

1554
01:18:38,270 --> 01:18:40,206
FLEXIBILITY AND HOW THAT MIGHT 

1555
01:18:40,206 --> 01:18:46,812
INTACT THE REST OF THE BCR 

1556
01:18:46,812 --> 01:18:49,381
DOMAINS.

1557
01:18:49,381 --> 01:18:52,184
AND THE MECHANISM IS DEFINED.

1558
01:18:52,184 --> 01:18:59,858
THE SECOND SET OF STRUCTURE AND 

1559
01:18:59,858 --> 01:19:04,396
HERE WE HAVE BOTH THE IGM AND 

1560
01:19:04,396 --> 01:19:09,468
IGG CLASS BCR AND BOTH ADOPT THE

1561
01:19:09,468 --> 01:19:10,569
SAME ORGANIZATION THAT WAS 

1562
01:19:10,569 --> 01:19:14,273
PREVIOUSLY REPORTED FOR THE 

1563
01:19:14,273 --> 01:19:24,450
MOUSE BCR.

1564
01:19:26,952 --> 01:19:30,489
WE GET ATOMIC INTERACTION AT THE

1565
01:19:30,489 --> 01:19:32,258
DOMAIN AND THE TRANS MEMBRANE 

1566
01:19:32,258 --> 01:19:33,692
DOMAIN.

1567
01:19:33,692 --> 01:19:35,894
SO THE OVER ALL ORGANIZATION OF 

1568
01:19:35,894 --> 01:19:38,230
THE BCR WAS SIMILAR FOR BOTH THE

1569
01:19:38,230 --> 01:19:40,733
IGG AS WELL AS THE IGM CLASS AND

1570
01:19:40,733 --> 01:19:46,038
AGAIN THERE IS NO INFORMATION 

1571
01:19:46,038 --> 01:19:49,942
ABOUT THE FAB AND THUS WE KNOW 

1572
01:19:49,942 --> 01:19:52,177
THERE WAS NO DESCRIPTION OF THE 

1573
01:19:52,177 --> 01:19:56,515
BCR DYNAMICS THAT RESULTS FROM 

1574
01:19:56,515 --> 01:19:59,752
THE OBSERVED FLEXIBILITY OF THE 

1575
01:19:59,752 --> 01:20:00,052
FAB.

1576
01:20:00,052 --> 01:20:00,953
ONE KEY INFORMATION THAT CAME 

1577
01:20:00,953 --> 01:20:06,025
FROM THAT STUDY WAS A KEY 

1578
01:20:06,025 --> 01:20:09,495
DIFFERENCE BETWEEN THE IGG AND 

1579
01:20:09,495 --> 01:20:14,133
IGM BCR CLASS.

1580
01:20:14,133 --> 01:20:15,467
THAT DIFFERENCE COMES FROM HOW 

1581
01:20:15,467 --> 01:20:18,270
THE FC DOMAIN INTERACTED OR 

1582
01:20:18,270 --> 01:20:19,838
ASSOCIATED WITH AG ALPHA.

1583
01:20:19,838 --> 01:20:23,275
SO IF YOU LOOK HERE IN THE IG 

1584
01:20:23,275 --> 01:20:27,446
AND BCR AND THIS IS THE IGL IN 

1585
01:20:27,446 --> 01:20:30,182
BLUE YOU CAN SEE IT'S STACKED 

1586
01:20:30,182 --> 01:20:33,485
TIGHTLY WITH THE FC DOMAIN 

1587
01:20:33,485 --> 01:20:36,188
UNLIKE WHAT WE HAVE SEEN THE IGG

1588
01:20:36,188 --> 01:20:42,728
BCR STRUCTURE.

1589
01:20:42,728 --> 01:20:45,564
WE THINK THIS IS PERHAPS 

1590
01:20:45,564 --> 01:20:46,398
RELEVANT FOR SIGNALLING BECAUSE 

1591
01:20:46,398 --> 01:20:50,035
WHEN WE LOOK AT SIGNALLING AND 

1592
01:20:50,035 --> 01:20:52,237
I'M SHOWING DATA FOR CALCIUM 

1593
01:20:52,237 --> 01:20:58,210
FLUX RESPONSES FOR 2G12IGG CELLS

1594
01:20:58,210 --> 01:21:03,015
AND IGM WE FOUND THE SIGNALLING 

1595
01:21:03,015 --> 01:21:05,150
WAS DIFFERENT FOR THE SAME PANEL

1596
01:21:05,150 --> 01:21:13,225
OF PRITRIMER AND OBSERVED WEAKE 

1597
01:21:13,225 --> 01:21:15,828
SIGNALLING IN THE IGG EXPRESSING

1598
01:21:15,828 --> 01:21:16,528
CELLS COMPARED TO THOSE 

1599
01:21:16,528 --> 01:21:21,734
EXPRESSING IGM.

1600
01:21:21,734 --> 01:21:25,971
SO WE DID SINGLE CRYO EM STUDIES

1601
01:21:25,971 --> 01:21:30,642
TO GET THE STRUCTURE OF BOTH THE

1602
01:21:30,642 --> 01:21:35,080
I SHAPED 2G12 AS WELL AS THE CD4

1603
01:21:35,080 --> 01:21:37,683
BINDING SITE THAT ADOPTS THE 

1604
01:21:37,683 --> 01:21:48,193
CANONICAL SHAPE CONFIGURATION.

1605
01:21:50,462 --> 01:21:55,801
WE WERE ABLE TO LOOK AT THE 

1606
01:21:55,801 --> 01:21:59,438
STRUCTURE AND THE STRUCTURE CAME

1607
01:21:59,438 --> 01:22:01,840
FROM A SMALL SUBSET OF 

1608
01:22:01,840 --> 01:22:02,107
PARTICLES.

1609
01:22:02,107 --> 01:22:06,512
THE MAJORITY SHOWED STRUCTURES 

1610
01:22:06,512 --> 01:22:08,680
WHERE WE COULD VISUALIZE THE 

1611
01:22:08,680 --> 01:22:11,016
SIGNALLING COMPONENT AND THE FC 

1612
01:22:11,016 --> 01:22:14,286
ARE THE FIVE ASSOCIATED WITH THE

1613
01:22:14,286 --> 01:22:17,089
FC.

1614
01:22:17,089 --> 01:22:23,395
THIS IS TRUE FOR CG12 AND IGM 

1615
01:22:23,395 --> 01:22:24,997
AND ANOTHER SHOWING OF THE FAB 

1616
01:22:24,997 --> 01:22:26,965
AND TOOK THE STRATEGY OF 

1617
01:22:26,965 --> 01:22:27,633
RECONSTRUCTING IT BY USING 

1618
01:22:27,633 --> 01:22:31,503
INFORMATION THAT CAME FROM THE 

1619
01:22:31,503 --> 01:22:34,206
MAJORITY OF THE PARTICLE WITH 

1620
01:22:34,206 --> 01:22:43,715
THE BOTTOM PART OF THE BCRs AND 

1621
01:22:43,715 --> 01:22:47,352
COMPLETED THE STRUCTURE.

1622
01:22:47,352 --> 01:22:49,588
CONSISTENT WITH THE PUBLISHED 

1623
01:22:49,588 --> 01:22:51,690
BCR STRUCTURE AND HERE BGR12 AND

1624
01:22:51,690 --> 01:22:58,163
THE ASYMMETRY OF THE BETA IN ITS

1625
01:22:58,163 --> 01:23:00,432
ORIENTATION WITH THE 

1626
01:23:00,432 --> 01:23:00,799
IMMUNOGLOBULIN.

1627
01:23:00,799 --> 01:23:01,767
THAT WAS CONSISTENTLY SEEN IN 

1628
01:23:01,767 --> 01:23:11,610
BOTH OF OUR STRUCTURES.

1629
01:23:11,610 --> 01:23:15,881
UNLIKE WHAT WAS REPORTED IN THE 

1630
01:23:15,881 --> 01:23:16,982
STRUCK STTURES WE OBSERVED KEY 

1631
01:23:16,982 --> 01:23:17,282
DIFFERENCES.

1632
01:23:17,282 --> 01:23:24,389
I'M SHOWING THE IGG BCR 

1633
01:23:24,389 --> 01:23:25,824
PUBLISHED STRUCTURE WITH OUR 

1634
01:23:25,824 --> 01:23:36,235
RECONSTRUCTED STRUCTURE.

1635
01:23:58,457 --> 01:24:00,158
HERE YOU CAN SEE THE VARIATION 

1636
01:24:00,158 --> 01:24:02,661
FROM THE PUBLISHED STRUCTURE ON 

1637
01:24:02,661 --> 01:24:04,229
THE IG ALPHA DOMAIN AND HERE IS 

1638
01:24:04,229 --> 01:24:06,098
IT THE LOOP THAT CONNECTS TO THE

1639
01:24:06,098 --> 01:24:09,701
TRANS MEMBRANE DOMAIN AND ALSO 

1640
01:24:09,701 --> 01:24:16,041
WITHIN THE BCR WE OBSERVED MUCH 

1641
01:24:16,041 --> 01:24:17,576
VARIABILITY WHEN WE COMPARED OR 

1642
01:24:17,576 --> 01:24:19,611
ALIGNED OUR STRUCTURE TO THOSE 

1643
01:24:19,611 --> 01:24:22,214
OF THE IGG STRUCTURE THAT WAS 

1644
01:24:22,214 --> 01:24:29,054
PUBLISHED.

1645
01:24:29,054 --> 01:24:34,426
WHEN WE LOOKED AT THE 

1646
01:24:34,426 --> 01:24:38,730
INTERACTION OF THE C LAMBDA 3 

1647
01:24:38,730 --> 01:24:43,068
DOMAIN WE FOUND THE 

1648
01:24:43,068 --> 01:24:46,204
IMMUNOGLOBULIN HAS STRONG 

1649
01:24:46,204 --> 01:24:48,140
INTERACTIONS WITH THE DOMAIN AND

1650
01:24:48,140 --> 01:24:51,143
THIS IS CONSISTENT WITH WHAT WAS

1651
01:24:51,143 --> 01:24:52,110
DESCRIBED IN THE IGG PUBLISHED 

1652
01:24:52,110 --> 01:24:56,481
STRUCTURE.

1653
01:24:56,481 --> 01:24:57,883
AND THESE STRONGER INTERACTIONS 

1654
01:24:57,883 --> 01:25:00,786
WERE NOT OBSERVED WHEN WE LOOKED

1655
01:25:00,786 --> 01:25:03,722
AT THE INTERACTIONS BETWEEN THE 

1656
01:25:03,722 --> 01:25:09,194
SIGNALLING COMPONENT AND THE FC 

1657
01:25:09,194 --> 01:25:16,268
AND AGAIN THIS WAS AN INDCAT

1658
01:25:16,268 --> 01:25:19,338
INDCATION -- INDICATION OR 

1659
01:25:19,338 --> 01:25:20,872
SUPPORTS THE MALLEABILITY OF THE

1660
01:25:20,872 --> 01:25:22,574
STRUCTURE.

1661
01:25:22,574 --> 01:25:24,776
WE WERE ABLE TO GET 

1662
01:25:24,776 --> 01:25:26,778
RECONSTRUCTED MAPS OF THE 

1663
01:25:26,778 --> 01:25:29,147
STRUCTURE AND THESE COMPOSITE 

1664
01:25:29,147 --> 01:25:31,416
MAPS VARIED IN THEIR RELATIVE 

1665
01:25:31,416 --> 01:25:34,753
ORIENTATION OF THE FACT TO THE 

1666
01:25:34,753 --> 01:25:34,920
FC.

1667
01:25:34,920 --> 01:25:38,256
WHEN YOU FIT THEM ALL TOGETHER 

1668
01:25:38,256 --> 01:25:43,428
YOU CAN SEE THE FAB ARE 

1669
01:25:43,428 --> 01:25:45,497
POSITIONED WITH A LOT OF 

1670
01:25:45,497 --> 01:25:48,700
VARIATIONS WITH RESPECT TO HOW 

1671
01:25:48,700 --> 01:25:50,302
THEY ORIENTED WITH THE FC AND 

1672
01:25:50,302 --> 01:25:51,336
REST OF THE BCR.

1673
01:25:51,336 --> 01:25:55,607
YOU CAN GET A SENSE OF THE 

1674
01:25:55,607 --> 01:25:58,243
EXTEND MUCH THIS VARIATION IF 

1675
01:25:58,243 --> 01:26:02,614
YOU LOOK AT THE DISTANCE WHICH 

1676
01:26:02,614 --> 01:26:13,158
RANGES FROM 132 TO 167 ENSTRON 

1677
01:26:26,238 --> 01:26:31,543
AND YOU SEE HOW IT'S ORIENTED 

1678
01:26:31,543 --> 01:26:42,087
RELATIVE TO THE FC HERE WE GOT 

1679
01:26:44,589 --> 01:26:46,091
GOOD DENSITY OF THE FAB AND WE 

1680
01:26:46,091 --> 01:26:47,993
SEE FLEXIBILITY IN THE 

1681
01:26:47,993 --> 01:26:51,163
ORIENTATION OF THE FAB AS 

1682
01:26:51,163 --> 01:26:57,469
EXPECTED ADOPTS THE CANONICAL Y 

1683
01:26:57,469 --> 01:26:59,471
CONFIGURATION AND WE COULD SEE 

1684
01:26:59,471 --> 01:27:03,008
WHERE THE VARIABILITY OF WHERE 

1685
01:27:03,008 --> 01:27:06,511
IT OCCURS AND THIS IS THE FAB FC

1686
01:27:06,511 --> 01:27:16,788
HINGE REGION AND THEN WE CR CAN 

1687
01:27:16,788 --> 01:27:22,260
CME1 AND 2 AND DISTAL TO IT IN 

1688
01:27:22,260 --> 01:27:23,862
THE CME 4 WE SEE A LOT OF 

1689
01:27:23,862 --> 01:27:24,162
FLEXIBILITY.

1690
01:27:24,162 --> 01:27:28,366
IT'S NOT ONLY COMING FROM THE 

1691
01:27:28,366 --> 01:27:32,204
FAB BUT WE ARE SEEING 

1692
01:27:32,204 --> 01:27:32,971
FLEXIBILITY WITHIN THE FC 

1693
01:27:32,971 --> 01:27:36,274
DOMAIN.

1694
01:27:36,274 --> 01:27:39,745
NOW, IF YOU PUT SIDE BY SIDE TO 

1695
01:27:39,745 --> 01:27:41,913
WHAT THE PUBLISHED STRUCTURE 

1696
01:27:41,913 --> 01:27:44,349
LOOKS LIKE AND SO THIS IS NOW 

1697
01:27:44,349 --> 01:27:49,054
THE IGM THE SAME CLASS AS THE 

1698
01:27:49,054 --> 01:27:50,622
CH31IGM WE MAPPED AROUND YOU CAN

1699
01:27:50,622 --> 01:27:54,059
SEE THE STRUCTURE THAT'S 

1700
01:27:54,059 --> 01:27:56,061
PUBLISHED IS AN ABRIDGED 

1701
01:27:56,061 --> 01:28:00,198
STRUCTURE AND STANDS TALL AND 

1702
01:28:00,198 --> 01:28:03,401
FULLY EXTENDED FORM FOR THE 

1703
01:28:03,401 --> 01:28:07,639
PLASMA MEMBRANE WHEREAS THE 

1704
01:28:07,639 --> 01:28:09,307
THREE COMPOSITE MAPS SHOW 

1705
01:28:09,307 --> 01:28:11,476
WIDESPREAD VARIATION AT THE FAB 

1706
01:28:11,476 --> 01:28:13,078
FROM THE FAB DOMAIN ALL THE WAY 

1707
01:28:13,078 --> 01:28:23,522
DOWN EXTENDING TO THE FC.

1708
01:28:25,390 --> 01:28:28,226
SO THIS LEADS US TO OUR CURRENT 

1709
01:28:28,226 --> 01:28:30,061
MODEL WHICH IS DISTINCT FROM THE

1710
01:28:30,061 --> 01:28:31,696
PUBLISHED MODEL AND WE ARE 

1711
01:28:31,696 --> 01:28:36,835
HYPOTHESIZING THE BENDING OF THE

1712
01:28:36,835 --> 01:28:39,638
IMMUNOGLOBULIN UNIT ALTERS THE 

1713
01:28:39,638 --> 01:28:41,273
SIGNALLING COMPONENT AND AN 

1714
01:28:41,273 --> 01:28:44,042
IMPLICATION OF THIS IS WHEN 

1715
01:28:44,042 --> 01:28:48,146
ANTIGEN BINDS BASED ON THIS 

1716
01:28:48,146 --> 01:28:50,081
MODEL WE HYPOTHESIZE THE BINDING

1717
01:28:50,081 --> 01:28:51,149
WILL INDUCE SOME CHANGES TO 

1718
01:28:51,149 --> 01:28:53,819
THIS.

1719
01:28:53,819 --> 01:28:56,288
SO THAT'S SOMETHING WE ARE 

1720
01:28:56,288 --> 01:29:00,158
CURRENTLY WORKING TO UNDERSTAND.

1721
01:29:00,158 --> 01:29:04,162
IN CONCLUSION WE ARE IN 

1722
01:29:04,162 --> 01:29:09,668
AGREEMENT THAT CH31 AND THE BCR 

1723
01:29:09,668 --> 01:29:12,370
STRUCTURES SHOW ASYMMETRIC 

1724
01:29:12,370 --> 01:29:12,704
ORGANIZATION.

1725
01:29:12,704 --> 01:29:16,274
THE WITH I SHAPED ECTODOMAIN 

1726
01:29:16,274 --> 01:29:18,243
SHOW CONFIRMATIONAL VARIABILITY 

1727
01:29:18,243 --> 01:29:21,213
THAT INDICATE HIGHLY DYNAMIC 

1728
01:29:21,213 --> 01:29:21,513
STRUCTURES.

1729
01:29:21,513 --> 01:29:22,581
WE OBSERVED DISTINCT 

1730
01:29:22,581 --> 01:29:26,518
CONFIRMATIONS OF THE FAB REGIONS

1731
01:29:26,518 --> 01:29:28,253
AS WELL AS ADDITIONAL HINGE 

1732
01:29:28,253 --> 01:29:29,888
REGION WITHIN THE FC THAT 

1733
01:29:29,888 --> 01:29:30,488
CONTRIBUTES TO FURTHER FAB 

1734
01:29:30,488 --> 01:29:32,390
DYNAMICS.

1735
01:29:32,390 --> 01:29:34,926
SO WE HYPOTHESIZED THAT THE 

1736
01:29:34,926 --> 01:29:36,761
HINGE POINTS WE'VE IDENTIFIED IN

1737
01:29:36,761 --> 01:29:46,271
OUR STRUCTURES ARE KEY FOR BCR 

1738
01:29:46,271 --> 01:29:47,505
ALLOSTERIC SIGNALLING AND I 

1739
01:29:47,505 --> 01:29:48,607
ACKNOWLEDGE THE LARGE NUMBER OF 

1740
01:29:48,607 --> 01:29:49,441
PEOPLE WHO CONTRIBUTED TO THIS 

1741
01:29:49,441 --> 01:29:49,641
WORK.

1742
01:29:49,641 --> 01:29:59,818
THANK YOU.

1743
01:30:05,490 --> 01:30:07,626
>> THANKS AGAIN AND I
WOULD LIKE

1744
01:30:07,626 --> 01:30:11,429
TO THANKS DAVID FOR THE 

1745
01:30:11,429 --> 01:30:14,332
OPPORTUNITY TO PRESENT RESULTS 

1746
01:30:14,332 --> 01:30:16,301
FROM OUR COMPUTATIONAL WORK.

1747
01:30:16,301 --> 01:30:18,303
IT FOCUSES ON LOOKING AT 

1748
01:30:18,303 --> 01:30:19,337
ANTIGENS AND WHAT HAPPENS 

1749
01:30:19,337 --> 01:30:20,505
OUTSIDE AND HOW DO YOU STABILIZE

1750
01:30:20,505 --> 01:30:23,141
THEM AND OUR INTEREST WAS TO SEE

1751
01:30:23,141 --> 01:30:25,010
HOW THEY GET PRESENTED INSIDE 

1752
01:30:25,010 --> 01:30:26,645
WHEN GIVEN AS A VACCINE ANTIGEN 

1753
01:30:26,645 --> 01:30:30,181
FOR EXAMPLE.

1754
01:30:30,181 --> 01:30:33,551
AND NORMALLY WHEN YOU HAVE GIVEN

1755
01:30:33,551 --> 01:30:39,190
VACCINE ANTIGEN IT'S BEEN TAKEN 

1756
01:30:39,190 --> 01:30:41,393
UP PART NEF IMMUNE AND THEY CAN 

1757
01:30:41,393 --> 01:30:45,196
PRESENT TO THE BCR OR THE 

1758
01:30:45,196 --> 01:30:48,800
ANTIGEN CAN BE PRESENTED TO THE 

1759
01:30:48,800 --> 01:30:51,369
BCR AS YOU HEARD FROM MUNIR AND 

1760
01:30:51,369 --> 01:30:53,071
THEN TO THE PLASMA CELL 

1761
01:30:53,071 --> 01:30:53,438
DIFFERENTIATION.

1762
01:30:53,438 --> 01:30:56,374
THE KEY COMPONENTS IS THE CELL 

1763
01:30:56,374 --> 01:31:01,780
MEDIATED IMMUNITY WHICH IS HOW 

1764
01:31:01,780 --> 01:31:06,284
YOU'RE GETTING THE T CELL HELP 

1765
01:31:06,284 --> 01:31:08,720
TOE INDUCE THE BCR IN THE CD8 T 

1766
01:31:08,720 --> 01:31:10,588
CELLS AND OUR FOCUS IS SEEING 

1767
01:31:10,588 --> 01:31:13,758
HOW THE ANTIGENS ARE GETTING 

1768
01:31:13,758 --> 01:31:16,061
RECOGNIZED BY THE B CELL AND T 

1769
01:31:16,061 --> 01:31:21,466
CELL AS A VACCINE ANTIGEN AND WE

1770
01:31:21,466 --> 01:31:28,506
LOOKED AT THE T CELL REPLICATION

1771
01:31:28,506 --> 01:31:30,976
A

1772
01:31:30,976 --> 01:31:33,044
AND YOU'LL NOTICE FROM WHAT YOU 

1773
01:31:33,044 --> 01:31:35,714
HEARD FROM MUNIR THEY'RE MODULAR

1774
01:31:35,714 --> 01:31:39,184
AND IT BINDS BUT THERE'S A 

1775
01:31:39,184 --> 01:31:47,759
SIGNALLING THAT IS DIFFERENT AND

1776
01:31:47,759 --> 01:31:54,866
THERE'S A SEQUENCE AND TALK 

1777
01:31:54,866 --> 01:31:56,401
ABOUT THE SIGNALLING.

1778
01:31:56,401 --> 01:31:58,670
WHEN WE STARTED THIS WORK I 

1779
01:31:58,670 --> 01:31:59,738
THINK SOME TIME BACK -- LET ME 

1780
01:31:59,738 --> 01:32:09,848
GO BACK TO THIS ONE.

1781
01:32:09,848 --> 01:32:11,149
WHEN WE HAVE THE DIFFERENT 

1782
01:32:11,149 --> 01:32:13,385
MEMBRANES WE FOUND WHEN YOU HAVE

1783
01:32:13,385 --> 01:32:17,722
A DISORDERED MEMBRANE THEY GET 

1784
01:32:17,722 --> 01:32:20,692
BURIED INSIDE BUT WHEN YOU GO IN

1785
01:32:20,692 --> 01:32:23,094
A DIFFERENT MEMBRANE LIKE A 

1786
01:32:23,094 --> 01:32:25,397
LIPID MEMBRANE THEY GET EXPOSED.

1787
01:32:25,397 --> 01:32:28,366
SO THIS LED US TO PROPOSE A 

1788
01:32:28,366 --> 01:32:31,536
MODEL WHERE MAYBE WHEN YOU HAVE 

1789
01:32:31,536 --> 01:32:33,371
THE SIGNALLING MOTIF IN 

1790
01:32:33,371 --> 01:32:35,206
DIFFERENT REGIONS THEY'RE HIDDEN

1791
01:32:35,206 --> 01:32:38,276
AND THEY COME AND MOVE ON TO A 

1792
01:32:38,276 --> 01:32:38,910
DIFFERENT LOCALIZATION POINT IN 

1793
01:32:38,910 --> 01:32:42,280
THE MEMBRANE DOMAIN AND THEY 

1794
01:32:42,280 --> 01:32:43,548
DECIDE TO GET EXPOSED AND LEAD 

1795
01:32:43,548 --> 01:32:47,118
TO A DOWN STREAM SIGNALLING.

1796
01:32:47,118 --> 01:32:48,753
AND I THINK SOME OF THE WORK HAS

1797
01:32:48,753 --> 01:32:55,660
BEEN SUPPORTED EXPERIMENTALLY.

1798
01:32:55,660 --> 01:33:00,365
SO TYPICALLY WHEN PEOPLE TALK 

1799
01:33:00,365 --> 01:33:03,468
ABOUT HOW THE CELLULAR BINDING 

1800
01:33:03,468 --> 01:33:05,937
LEAD TO INTRASIGNALLING 

1801
01:33:05,937 --> 01:33:07,906
DIFFERENT MODELS HAVE BEEN 

1802
01:33:07,906 --> 01:33:11,309
PROPOSED SOME CONFIRMATIONAL 

1803
01:33:11,309 --> 01:33:13,945
CHANGE OR THE EMERGING CONCEPT 

1804
01:33:13,945 --> 01:33:18,283
SEEMS TO BE THAT THE MEMBRANES 

1805
01:33:18,283 --> 01:33:20,618
CAN PLAY AN ACTIVE ROLE AND 

1806
01:33:20,618 --> 01:33:21,519
SOMETHING RELEVANT HAPPENS TO 

1807
01:33:21,519 --> 01:33:22,620
BCR AND I DON'T NEED TO GO IN 

1808
01:33:22,620 --> 01:33:25,223
THE DETAILS OF IT.

1809
01:33:25,223 --> 01:33:27,625
I THINK AS MUNIR MENTIONED 

1810
01:33:27,625 --> 01:33:29,661
PEOPLE THOUGHT IT WAS A 

1811
01:33:29,661 --> 01:33:30,728
SYMMETRIC COMPLEX BUT TURNED OUT

1812
01:33:30,728 --> 01:33:32,397
TO BE THE STRUCTURES CONFIRM 

1813
01:33:32,397 --> 01:33:34,699
WITH ASYMMETRIC.

1814
01:33:34,699 --> 01:33:37,435
SO ONE OF THE GOALS OF THE 

1815
01:33:37,435 --> 01:33:40,338
COMPUTATIONAL WORK IS THE EFFECT

1816
01:33:40,338 --> 01:33:45,710
OF HAVING AN ASYMMETRY IN THE 

1817
01:33:45,710 --> 01:33:49,647
CONTEXT OF MEMBRANE AND THOUGH 

1818
01:33:49,647 --> 01:33:51,282
EVERYONE HAS KNOWN FOR A LONG 

1819
01:33:51,282 --> 01:33:54,219
TIME THE BCR IS EFFECTIVE FOR 

1820
01:33:54,219 --> 01:33:56,387
IMMUNITY WE DON'T KNOW THE 

1821
01:33:56,387 --> 01:33:56,688
MECHANISMS.

1822
01:33:56,688 --> 01:34:02,293
SOME MODELS HAVE BEEN PROPOSED 

1823
01:34:02,293 --> 01:34:03,128
WHERE THE ANTIGEN BINDS AND 

1824
01:34:03,128 --> 01:34:04,562
CREATE CROSS LINKING AND LEADS 

1825
01:34:04,562 --> 01:34:05,597
TO SOME SORT OF SIGNALLING.

1826
01:34:05,597 --> 01:34:11,136
AND RECENTLY THERE WAS A 

1827
01:34:11,136 --> 01:34:12,704
DIFFERENT CALLED DISSOCIATION 

1828
01:34:12,704 --> 01:34:14,272
ACTIVATION STATE AND IT STAYS 

1829
01:34:14,272 --> 01:34:15,507
TOGETHER AND SOME ANTIGEN BINDS 

1830
01:34:15,507 --> 01:34:18,343
AND SOME SHIFTS INTO A MORE OF 

1831
01:34:18,343 --> 01:34:22,714
AN ACTIVE BOND.

1832
01:34:22,714 --> 01:34:28,353
AND WE'LL LOOK AT THE 

1833
01:34:28,353 --> 01:34:28,653
SIMULATIONS.

1834
01:34:28,653 --> 01:34:30,889
SO WE SELECTED THE STIMULATION 

1835
01:34:30,889 --> 01:34:31,990
WORK ON THE BCR COMPLEX BECAUSE 

1836
01:34:31,990 --> 01:34:34,425
IT'S A WAY WE CAN CONNECT TO THE

1837
01:34:34,425 --> 01:34:40,765
WORK GOING ON AT DUKE.

1838
01:34:40,765 --> 01:34:51,309
AND MODEL THE MONOMER AND AND WE

1839
01:34:56,214 --> 01:34:59,584
IS ASSEMBLED THE COMPLEX B CELL 

1840
01:34:59,584 --> 01:35:01,085
LIKE BILAYER.

1841
01:35:01,085 --> 01:35:04,689
TODAY I THINK I'LL TALK MAINLY 

1842
01:35:04,689 --> 01:35:06,591
ABOUT THE OUTCOME OF THE 

1843
01:35:06,591 --> 01:35:10,762
SIMULATIONS FROM THIS KIND OF A 

1844
01:35:10,762 --> 01:35:13,164
COMPLEX WHERE THE B CELL 

1845
01:35:13,164 --> 01:35:16,100
MEMBRANE WAS CAPTURED WITH THE 

1846
01:35:16,100 --> 01:35:17,335
DIFFERENT LIPID COMPOSITION IN 

1847
01:35:17,335 --> 01:35:20,038
AN EXPERIMENTAL PAPER.

1848
01:35:20,038 --> 01:35:22,340
WE TRIED TO USE THE COMBINATION 

1849
01:35:22,340 --> 01:35:25,510
OF LIPIDS AND LOOK AT THE EFFECT

1850
01:35:25,510 --> 01:35:27,579
OF ANTIGEN BOUND VERSUS NOT 

1851
01:35:27,579 --> 01:35:30,582
BOUND AND SEE WHETHER WE CAN SEE

1852
01:35:30,582 --> 01:35:33,351
ANYTHING MORE IN THE BCR 

1853
01:35:33,351 --> 01:35:33,651
ACTIVATION.

1854
01:35:33,651 --> 01:35:40,358
WE FIND FOUR INTERESTING 

1855
01:35:40,358 --> 01:35:43,294
DYNAMICAL MOTIONS EVENTS FROM 

1856
01:35:43,294 --> 01:35:44,062
SIMULATION AND IT'S SOMETHING 

1857
01:35:44,062 --> 01:35:46,030
I'LL GO THROUGH IN THE NEXT FEW 

1858
01:35:46,030 --> 01:35:46,331
SLIDES.

1859
01:35:46,331 --> 01:35:48,733
ONE OF THE FIRST THINGS WE 

1860
01:35:48,733 --> 01:35:51,236
NOTICED ANTIGEN INDUCES CHANGES 

1861
01:35:51,236 --> 01:35:53,838
ACROSS THE WHOLE BCR.

1862
01:35:53,838 --> 01:35:55,106
SO HERE WE COMPARE THE 

1863
01:35:55,106 --> 01:36:04,215
COMPLEXABILITY OF  BOUND AND 

1864
01:36:04,215 --> 01:36:06,517
UNBOUND AND THERE'S A REGION 

1865
01:36:06,517 --> 01:36:17,028
WHERE THE BOUND REGION GETS 

1866
01:36:19,230 --> 01:36:22,300
RIGIDIFIED AND THE SECOND THING 

1867
01:36:22,300 --> 01:36:32,810
WE NOTICED AND THEY COME MORE 

1868
01:36:38,449 --> 01:36:40,385
TOWARDS ALONG THE Z AXIS IF YOU 

1869
01:36:40,385 --> 01:36:42,687
LOOK AT THE CASE HERE THE ANGLE 

1870
01:36:42,687 --> 01:36:46,090
REDUCES IN ALL THE TRANS 

1871
01:36:46,090 --> 01:36:51,462
MEMBRANE HELLISMSS  -- HELICES 

1872
01:36:51,462 --> 01:36:55,767
AND THE IGL BETA.

1873
01:36:55,767 --> 01:36:58,269
THEN THE THIRD PART WE OBSERVED 

1874
01:36:58,269 --> 01:37:01,072
IS IF YOU FOCUS ONLY ON THE 

1875
01:37:01,072 --> 01:37:03,574
SIGNALLING MOTIF THE ANTIGEN 

1876
01:37:03,574 --> 01:37:05,376
BINDING SEEMS TO REDUCE SOME 

1877
01:37:05,376 --> 01:37:09,847
INTERACTIONS MAINLY IN THE 

1878
01:37:09,847 --> 01:37:11,215
REGIONS OF THIS NEAR THE 

1879
01:37:11,215 --> 01:37:14,285
MEMBRANE REGION AND INCREASED 

1880
01:37:14,285 --> 01:37:17,121
CONTEXT IN THE REGIONS WHICH ARE

1881
01:37:17,121 --> 01:37:23,061
LIKE THE EXTRA CELLULAR AND WHAT

1882
01:37:23,061 --> 01:37:23,861
CAPTURED IN THIS CONTACT 

1883
01:37:23,861 --> 01:37:27,432
FREQUENCY MAP YOU SEE.

1884
01:37:27,432 --> 01:37:29,367
AND THERE'S ONE OTHER ASPECT WE 

1885
01:37:29,367 --> 01:37:31,402
WANTED TO SEE BECAUSE WE KNOW 

1886
01:37:31,402 --> 01:37:33,338
REGARDLESS OF WHATEVER THE MODEL

1887
01:37:33,338 --> 01:37:37,575
A CROSS LINK O

1888
01:37:37,575 --> 01:37:42,814
OOR DISASSOCI

1889
01:37:42,814 --> 01:37:44,782
OR DISASSOCIATION/ACTIVATION 

1890
01:37:44,782 --> 01:37:46,217
THEY NEED TO COME TOGETHER.

1891
01:37:46,217 --> 01:37:47,785
THIS COULD HAPPEN THROUGH MANY 

1892
01:37:47,785 --> 01:37:48,052
WAYS.

1893
01:37:48,052 --> 01:37:50,988
MAYBE THREES RE-ORIENTATION OF 

1894
01:37:50,988 --> 01:37:52,423
THE TRANS MEMBRANE REGION WHERE 

1895
01:37:52,423 --> 01:37:54,258
THINGS BECOME MORE STICKY AND 

1896
01:37:54,258 --> 01:37:56,928
PUT OTHER BCRs TOGETHER OR 

1897
01:37:56,928 --> 01:37:58,129
ALTERNATIVELY YOU COULD HAVE A 

1898
01:37:58,129 --> 01:38:00,565
CASE WHERE SOMETHING CHANGES 

1899
01:38:00,565 --> 01:38:02,300
BECAUSE THE TRANS MEMBRANE 

1900
01:38:02,300 --> 01:38:03,668
REGION CHANGES SOMEWHAT AND 

1901
01:38:03,668 --> 01:38:04,902
CHANGES TO LIPID COMPOSITION 

1902
01:38:04,902 --> 01:38:06,537
AROUND IT AND COULD LEAD TO A 

1903
01:38:06,537 --> 01:38:07,171
DIFFERENT LOCALIZATION IN THE 

1904
01:38:07,171 --> 01:38:09,640
MEMBRANE.

1905
01:38:09,640 --> 01:38:13,911
AGAIN JUST TO FIND OUT THESE ARE

1906
01:38:13,911 --> 01:38:16,080
SOME PAPERS OF QUESTIONS PEOPLE 

1907
01:38:16,080 --> 01:38:18,082
HAVE ASKED BEFORE.

1908
01:38:18,082 --> 01:38:20,485
AND WE WERE CURIOUS THAT I 

1909
01:38:20,485 --> 01:38:24,655
MENTIONED THE THIRD THING WHEN 

1910
01:38:24,655 --> 01:38:26,591
THE ANTIGEN BINDS IT BECOMES 

1911
01:38:26,591 --> 01:38:27,959
MORE STRAIGHT.

1912
01:38:27,959 --> 01:38:32,663
AND IN A MEMBRANE-BASED SYSTEM 

1913
01:38:32,663 --> 01:38:37,635
THAT CAUSES SOME HYDROTROPHIC 

1914
01:38:37,635 --> 01:38:39,737
MISMATCH AND THE TRENDS IS 

1915
01:38:39,737 --> 01:38:42,240
THEY'LL GO TO A MEMBRANE DOMAIN 

1916
01:38:42,240 --> 01:38:43,040
WHICH IS THICKER.

1917
01:38:43,040 --> 01:38:49,013
SO I THINK YOU HEARD FROM HUANG 

1918
01:38:49,013 --> 01:38:50,281
YESTERDAY WE HAVE DEVELOPED THE 

1919
01:38:50,281 --> 01:38:52,450
CROSS LINK AND THE LIPID MIXING 

1920
01:38:52,450 --> 01:38:54,051
WHEN YOU HAVE A COMPLEX LIPID IT

1921
01:38:54,051 --> 01:38:57,221
TAKES A LONG TIME TO MAKE SO WE 

1922
01:38:57,221 --> 01:39:02,260
TOOK THIS AND RAN EXTENSIVE 

1923
01:39:02,260 --> 01:39:06,164
SIMULATIONS.

1924
01:39:06,164 --> 01:39:09,934
AND WHAT WE ARE SHOWING HERE THE

1925
01:39:09,934 --> 01:39:12,136
COARSE-GRAINED SIMULATIONS AND 

1926
01:39:12,136 --> 01:39:15,573
WE'RE LOOKING AND ASKING THE 

1927
01:39:15,573 --> 01:39:19,644
QUESTION WITHIN THE SHORT REGION

1928
01:39:19,644 --> 01:39:24,215
ARE THE LIPIDS CHANGING WITH THE

1929
01:39:24,215 --> 01:39:28,386
ANTIGENS BOUND VERSUS NOT BOUND.

1930
01:39:28,386 --> 01:39:30,288
WE SEE SOME CHANGES WHEN ANTIGEN

1931
01:39:30,288 --> 01:39:34,292
IS BOUND VERSUS NOT BOUND.

1932
01:39:34,292 --> 01:39:38,262
WE SEE INCREASED IN CHOLESTEROL 

1933
01:39:38,262 --> 01:39:40,164
AND LIPIDS SOISHTED WITH MORE 

1934
01:39:40,164 --> 01:39:43,534
ORDER DOMAIN OR A THICKER 

1935
01:39:43,534 --> 01:39:43,768
DOMAIN.

1936
01:39:43,768 --> 01:39:45,269
THIS RAISES WHETHER THERE'S AN 

1937
01:39:45,269 --> 01:39:50,274
EFFECT WHERE THIS COULD LEAD TO 

1938
01:39:50,274 --> 01:39:55,313
SOME SORT OF LOCALIZATION AND IN

1939
01:39:55,313 --> 01:39:58,249
SUMMARY WHAT WE ARE FINDING OUT 

1940
01:39:58,249 --> 01:40:00,952
IS THAT WHEN THE ANTIGEN BINDS 

1941
01:40:00,952 --> 01:40:06,757
THERE SEEMS TO BE A DIFFERENTIAL

1942
01:40:06,757 --> 01:40:16,667
MOTION WITH ALLOSTERIC 

1943
01:40:16,667 --> 01:40:18,603
INTERACTIONS AND THIRD WE SEE 

1944
01:40:18,603 --> 01:40:20,505
THE ALTERATIONS ARE IN THE 

1945
01:40:20,505 --> 01:40:25,176
SIGNALLING MOTIF AND THE IDEA IS

1946
01:40:25,176 --> 01:40:30,214
THE SIGNALLING MOTIF MIGHT 

1947
01:40:30,214 --> 01:40:40,725
IMPACT THE ONE MOTIF EXPOSED.

1948
01:40:44,795 --> 01:40:50,601
AND AGAIN I THINK MOST THE WORK 

1949
01:40:50,601 --> 01:40:53,304
WAS DONE BY HUANG DO AND NATURAL

1950
01:40:53,304 --> 01:40:56,207
WAS INVOLVED IN THE 

1951
01:40:56,207 --> 01:40:57,208
COMPUTATIONAL BIOLOGY CENTER HIV

1952
01:40:57,208 --> 01:40:58,175
CENTER GROUP AT DUKE.

1953
01:40:58,175 --> 01:40:58,709
AND THAT'S ALL I HAVE.

1954
01:40:58,709 --> 01:41:08,886
THANK YOU.

1955
01:41:31,776 --> 01:41:32,977
>> I'LL MENTION WE ARE IN
THE 

1956
01:41:32,977 --> 01:41:34,278
CORE OF STRUCTURAL BIOLOGY AND 

1957
01:41:34,278 --> 01:41:38,282
ONE MANDATE IS TO PROVIDE 

1958
01:41:38,282 --> 01:41:40,318
SUPPORT IN TERMS OF MUTANT 

1959
01:41:40,318 --> 01:41:45,790
KNOWLEDGE THAT CAN BE APPLIED TO

1960
01:41:45,790 --> 01:41:48,659
THE BIOLOGY AND ONE WE'RE 

1961
01:41:48,659 --> 01:41:58,302
EXCITED ABOUT IS SINGLE  

1962
01:41:58,302 --> 01:42:03,808
CRYOELECTRON TOMOGRAPHY AND IN A

1963
01:42:03,808 --> 01:42:04,909
NUTSHELL WHAT THE TECHNOLOGY 

1964
01:42:04,909 --> 01:42:12,950
ALLOWS YOU TO DO IS TO LOOK AT 

1965
01:42:12,950 --> 01:42:13,918
LOOK AT STRUCTURAL BIOLOGY SO 

1966
01:42:13,918 --> 01:42:15,319
YOU CAN LOOK AT THE CELL.

1967
01:42:15,319 --> 01:42:16,921
IT'S TOO THICK TO GET THROUGH 

1968
01:42:16,921 --> 01:42:19,757
THE MICROSCOPE AND NEED TO THIN 

1969
01:42:19,757 --> 01:42:23,294
IT THROUGH THE PROCESS.

1970
01:42:23,294 --> 01:42:26,931
ONCE YOU HAVE IT THIN ENOUGH YOU

1971
01:42:26,931 --> 01:42:35,573
CAN TAKE PROJEBBICTIONS AND LOO 

1972
01:42:35,573 --> 01:42:38,342
FOR REPEATING FEATURES AND THAT 

1973
01:42:38,342 --> 01:42:39,644
COULD BE LIKE THE ENVELOPE OR 

1974
01:42:39,644 --> 01:42:41,145
ANY OTHER PROTEIN YOU HAVE IN 

1975
01:42:41,145 --> 01:42:45,816
THE CELL IN THIS CASE AND 

1976
01:42:45,816 --> 01:42:48,819
COMBINE THOSE VOLUMES BY USING 

1977
01:42:48,819 --> 01:42:51,288
TOMOGRAPHY TO GET INFORMATION 

1978
01:42:51,288 --> 01:42:51,689
FROM YOUR DATA.

1979
01:42:51,689 --> 01:42:55,192
TO GIVE A SENSE WHERE THE FIELD 

1980
01:42:55,192 --> 01:42:55,860
IS.

1981
01:42:55,860 --> 01:43:00,498
IT'S BEEN AROUND A LONG TIME BUT

1982
01:43:00,498 --> 01:43:02,433
RECENTLY HAS COME A LONG WAY IN 

1983
01:43:02,433 --> 01:43:04,301
TERMS OF TECHNOLOGY DEVELOPMENT.

1984
01:43:04,301 --> 01:43:06,837
THIS IS THE LAY OF WHAT CAN BE 

1985
01:43:06,837 --> 01:43:08,139
ACCOMPLISHED THESE DAYS.

1986
01:43:08,139 --> 01:43:11,776
SO IN THIS GRAPH YOU HAVE ON THE

1987
01:43:11,776 --> 01:43:13,344
X AXIS THE COMPLEXITY OF THE 

1988
01:43:13,344 --> 01:43:13,778
DIFFERENT SAMPLES.

1989
01:43:13,778 --> 01:43:18,049
AT THE BEGINNING ARE THE 

1990
01:43:18,049 --> 01:43:20,384
PURIFIED SAMPLE WHAT YOU 

1991
01:43:20,384 --> 01:43:23,988
TYPICALLY DO WITH CRYO EM AND 

1992
01:43:23,988 --> 01:43:26,290
YOU SEE IN VITRO PROTEIN LIKE 

1993
01:43:26,290 --> 01:43:32,396
HIV GAG AND THEN YOU MOVE NEXT 

1994
01:43:32,396 --> 01:43:37,702
STEP BACTERIAL SAMPLES AND TO 

1995
01:43:37,702 --> 01:43:42,640
THE RIGHT IS THE LARGER 

1996
01:43:42,640 --> 01:43:45,109
EUKARYOTIC CELLS TO THIN THE 

1997
01:43:45,109 --> 01:43:45,342
SAMPLE.

1998
01:43:45,342 --> 01:43:47,945
AND THE STRUCTURES ON THE GRAPH 

1999
01:43:47,945 --> 01:43:50,247
THE HIGHER THEY WILL THE HIGHER 

2000
01:43:50,247 --> 01:43:51,015
THE RESOLUTION.

2001
01:43:51,015 --> 01:43:53,684
FOR THE SAMPLES LIKE PURIFIED 

2002
01:43:53,684 --> 01:43:58,289
PROTEIN WE CAN GET PURIFIED 

2003
01:43:58,289 --> 01:44:01,225
RESOLUTION USING CRYO ET AND 

2004
01:44:01,225 --> 01:44:03,127
THEN AS YOU GO TO THE RIGHT TO 

2005
01:44:03,127 --> 01:44:05,429
THE MORE COMPLEX SAMPLES THE 

2006
01:44:05,429 --> 01:44:06,263
RESOLUTION SUFFERS A LITTLE BIT 

2007
01:44:06,263 --> 01:44:07,398
BUT WE'RE TRYING TO DEVELOP 

2008
01:44:07,398 --> 01:44:10,167
METHODS TO TRY TO IMPROVE THE 

2009
01:44:10,167 --> 01:44:12,236
FEATURES WE CAN SEE EVEN IN THE 

2010
01:44:12,236 --> 01:44:12,903
MORE COMPLEX SAMPLES.

2011
01:44:12,903 --> 01:44:16,474
ONE THING THAT IS IMPORTANT TO 

2012
01:44:16,474 --> 01:44:18,776
NOTE HERE IS THAT ONE PARAMETER 

2013
01:44:18,776 --> 01:44:19,677
THAT IS IMPORTANT IS THE NUMBER 

2014
01:44:19,677 --> 01:44:23,214
OF COPIES WE HAVE ACCESS TO AND 

2015
01:44:23,214 --> 01:44:27,318
OBVIOUSLY IN THE MORE COMPLEX 

2016
01:44:27,318 --> 01:44:30,588
SAMPLES AND HAD PHYSIOLOGICAL 

2017
01:44:30,588 --> 01:44:34,458
CONCENTRATIONS AND YOU CAN SEE 

2018
01:44:34,458 --> 01:44:36,861
THESE START TO GO DOWN AS YOU 

2019
01:44:36,861 --> 01:44:37,762
MOVE TO THE RIGHT.

2020
01:44:37,762 --> 01:44:39,897
HOPEFULLY IT GIVES AN OVERVIEW 

2021
01:44:39,897 --> 01:44:42,933
OF WHERE THE FIELD IS ON THINGS 

2022
01:44:42,933 --> 01:44:43,467
MOVING FORWARD IT'S VERY 

2023
01:44:43,467 --> 01:44:47,438
EXCITING.

2024
01:44:47,438 --> 01:44:50,674
ONE OF THE THINGS THAT HAS 

2025
01:44:50,674 --> 01:44:53,778
DRIVEN IMPROVEMENTS IS 

2026
01:44:53,778 --> 01:44:55,746
PREPARATION AND ALSO IMAGING AND

2027
01:44:55,746 --> 01:44:57,314
WHAT MY LAB HAS BEEN FOCUSSING 

2028
01:44:57,314 --> 01:44:59,283
ON IS ONCE YOU HAVE YOUR DATA 

2029
01:44:59,283 --> 01:45:00,684
HOW YOU IMPROVE THE DATA 

2030
01:45:00,684 --> 01:45:04,922
ANALYSIS TO GET THE MOST OUT OF 

2031
01:45:04,922 --> 01:45:05,289
YOUR SERIES.

2032
01:45:05,289 --> 01:45:08,592
AND YOU CAN SEE HERE THAT 

2033
01:45:08,592 --> 01:45:09,360
ACTUALLY THE COMBINATION OF 

2034
01:45:09,360 --> 01:45:13,731
DIFFERENT IMPROVEMENTS AND 

2035
01:45:13,731 --> 01:45:16,367
METHODS CAN TAKE YOU FROM WHAT 

2036
01:45:16,367 --> 01:45:19,904
IS ON THE LEFT THROUGH START 

2037
01:45:19,904 --> 01:45:20,671
SEEING SIDE CHAINS AND MODELLING

2038
01:45:20,671 --> 01:45:23,307
BECOMES A LOT EASIER.

2039
01:45:23,307 --> 01:45:25,543
SO JUST TO GIVE AN OVERVIEW.

2040
01:45:25,543 --> 01:45:31,949
IF YOU'RE FAMILIAR WITH CRYO EM 

2041
01:45:31,949 --> 01:45:34,451
CRYO ET HAS A FEW MORE STEPS AND

2042
01:45:34,451 --> 01:45:35,519
TRYING TO DESCRIBE THE 

2043
01:45:35,519 --> 01:45:36,487
COMPLEXITY OF THE PROCESS AND 

2044
01:45:36,487 --> 01:45:39,490
THERE'S MULTIPLE STEPS.

2045
01:45:39,490 --> 01:45:42,059
RIGHT NOW IT'S QUITE INVOLVED 

2046
01:45:42,059 --> 01:45:43,093
AND YOU NEED TO KNOW A LOT OF 

2047
01:45:43,093 --> 01:45:44,395
THE DIFFERENT TOOLS AND PIECE 

2048
01:45:44,395 --> 01:45:46,063
THEM TOGETHER IN THE RIGHT WAY 

2049
01:45:46,063 --> 01:45:47,031
TO GET A HIGH RESOLUTION 

2050
01:45:47,031 --> 01:45:48,799
STRUCTURE AT THE END.

2051
01:45:48,799 --> 01:45:51,635
SO WE RECENTLY WROTE A REVIEW 

2052
01:45:51,635 --> 01:45:52,937
WITH ONE OF MY STUDENTS WHO WILL

2053
01:45:52,937 --> 01:45:56,574
HAVE A POSTER IN THE AFTERNOON, 

2054
01:45:56,574 --> 01:46:00,311
JAVI, AND WE REVIEWED THE ENTIRE

2055
01:46:00,311 --> 01:46:08,953
LAY OF THE LAND IN SINGLE CRYO 

2056
01:46:08,953 --> 01:46:11,689
TOMOGRAPHY AND HOPEFULLY IT WILL

2057
01:46:11,689 --> 01:46:13,057
BE A GOOD RESOURCE FOR THE 

2058
01:46:13,057 --> 01:46:13,324
COMMUNITY.

2059
01:46:13,324 --> 01:46:14,925
AND IN ADDRESSING THE COMPLEXITY

2060
01:46:14,925 --> 01:46:19,496
WE WERE TRYING TO BUILD A LAYER 

2061
01:46:19,496 --> 01:46:22,099
ON TOP TO MAKE THE TECHNOLOGY 

2062
01:46:22,099 --> 01:46:24,134
AVAILABLE TO PEOPLE USING THIS 

2063
01:46:24,134 --> 01:46:26,470
STRUCTURES AND THE NAME OF THE 

2064
01:46:26,470 --> 01:46:29,273
PLATFORM WE DEVELOPED AND WE 

2065
01:46:29,273 --> 01:46:33,010
RELEASED LAST YEAR AND IT'S 

2066
01:46:33,010 --> 01:46:34,211
WEB-BASED AND INTUITIVE AND 

2067
01:46:34,211 --> 01:46:35,212
ALLOWS YOU ONCE YOU GET YOUR 

2068
01:46:35,212 --> 01:46:37,147
HANDS ON YOUR DATA YOU CAN PUSH 

2069
01:46:37,147 --> 01:46:40,184
IT THROUGH THIS PIPELINE AND 

2070
01:46:40,184 --> 01:46:45,022
ANALYZE AND SEE THE INTERMEDIATE

2071
01:46:45,022 --> 01:46:46,323
RESULTS AND HOPE IT WILL MAKE A 

2072
01:46:46,323 --> 01:46:48,225
BIG DIFFERENCE FOR THE FIELD AND

2073
01:46:48,225 --> 01:46:51,562
THIS IS OUR LAB THAT WAS 

2074
01:46:51,562 --> 01:46:52,363
FACILITATED TREMENDOUSLY AND THE

2075
01:46:52,363 --> 01:46:53,430
AMOUNT OF DATA WE CAN PROCESS 

2076
01:46:53,430 --> 01:46:56,433
AND THE TRAINING YOU NEED TO 

2077
01:46:56,433 --> 01:46:58,235
HAVE TO CONVERT RAW DATA TO HIGH

2078
01:46:58,235 --> 01:46:59,370
RESOLUTION STRUCTURES.

2079
01:46:59,370 --> 01:47:03,040
WE'RE EXCITED ABOUT THIS TOOL.

2080
01:47:03,040 --> 01:47:05,075
WE ARE HOPING IT WILL BE USEFUL 

2081
01:47:05,075 --> 01:47:07,912
IN A LOT OF THE PROJECTS WITHIN 

2082
01:47:07,912 --> 01:47:10,514
THE PROJECTS UP OUR CENTER BUT 

2083
01:47:10,514 --> 01:47:14,285
ALSO MORE BROADLY IN OTHER 

2084
01:47:14,285 --> 01:47:16,387
CASES.

2085
01:47:16,387 --> 01:47:18,289
SO ONE OF THE THINGS WE DO AND 

2086
01:47:18,289 --> 01:47:21,659
ONE EXAMPLE ON VIRUSES IS WE CAN

2087
01:47:21,659 --> 01:47:31,568
ANALYZE THE VULNERABILITY IN 

2088
01:47:31,568 --> 01:47:33,871
SITU AND WE LOOKED AT ONE TEST 

2089
01:47:33,871 --> 01:47:37,908
DATA SET ON THE RIBOSOME ON 

2090
01:47:37,908 --> 01:47:40,978
BACTERIAL CELLS AND CAN T CELL 

2091
01:47:40,978 --> 01:47:42,046
THE DIFFERENT FACTORS ATTACHED 

2092
01:47:42,046 --> 01:47:43,480
TO THE RIBOSOMES.

2093
01:47:43,480 --> 01:47:45,749
YOU CAN DO ALL THESE KINDS OF 

2094
01:47:45,749 --> 01:47:47,284
THINGS IN A MORE ROUTINE WAY 

2095
01:47:47,284 --> 01:47:54,158
SIMILAR TO WHAT YOU CAN DO WITH 

2096
01:47:54,158 --> 01:47:56,026
SINGLE CRYO.

2097
01:47:56,026 --> 01:47:57,995
AND THIS LEVERAGES THE POWER IN 

2098
01:47:57,995 --> 01:48:00,664
THE TOOLS AND THIS IS ON A 

2099
01:48:00,664 --> 01:48:06,270
STRUCTURE WE SAW RECENTLY IN 

2100
01:48:06,270 --> 01:48:12,676
COLLABORATION WITH THE IMMATURE 

2101
01:48:12,676 --> 01:48:18,215
CAPSID PROTEIN IN MISS CASE THEY

2102
01:48:18,215 --> 01:48:19,683
IMAGED THIS AND YOU CAN SEE HERE

2103
01:48:19,683 --> 01:48:24,888
IN THE UPPER LEFT YOU CAN SEE 

2104
01:48:24,888 --> 01:48:29,226
THE PROTEIN INSIDE THE BLPs AND 

2105
01:48:29,226 --> 01:48:31,562
ESSENTIALLY ONCE YOU TAKE THE 

2106
01:48:31,562 --> 01:48:40,504
TOMMO   -- TOMOGRAM AND CUT OUT 

2107
01:48:40,504 --> 01:48:41,271
THE POSITIONS WHERE YOU HAVE THE

2108
01:48:41,271 --> 01:48:42,973
GUT PROTEIN.

2109
01:48:42,973 --> 01:48:44,942
ALL THAT INVOLVES DOING A LOT OF

2110
01:48:44,942 --> 01:48:46,176
PREPARATIONS IN 3-D.

2111
01:48:46,176 --> 01:48:48,779
YOU NEED TO DO DETECTION, 

2112
01:48:48,779 --> 01:48:59,289
SEGMENTATION AND FIND ALL THE 

2113
01:48:59,590 --> 01:49:01,658
EXAMPLES AND WE HAVE THE SUB 

2114
01:49:01,658 --> 01:49:04,762
VOLUMES THAT CAPTURE A SMALL 

2115
01:49:04,762 --> 01:49:11,735
PIECE OF THE HEXAMERIC LATTICE 

2116
01:49:11,735 --> 01:49:13,303
AND YOU NEED THINGS FOR THE HIGH

2117
01:49:13,303 --> 01:49:14,271
RESOLUTION AND ONCE YOU HAVE 

2118
01:49:14,271 --> 01:49:17,141
THOSE YOU HAVE TO ALIGN THEM IN 

2119
01:49:17,141 --> 01:49:21,512
3-D AND GET A 3-D STRUCTURE AND 

2120
01:49:21,512 --> 01:49:23,280
THIS SHOWING DIFFERENT STEPS IN 

2121
01:49:23,280 --> 01:49:24,748
STAGES IN ORDER TO INCREASE 

2122
01:49:24,748 --> 01:49:28,218
RESOLUTION AND EVENTUALLY WE 

2123
01:49:28,218 --> 01:49:30,287
ENDED UP WITH A RESOLUTION 

2124
01:49:30,287 --> 01:49:38,262
RECONSTRUCTION YOU CAN SEE WITH 

2125
01:49:38,262 --> 01:49:39,897
THE CYAN PART HIGHLIGHTED.

2126
01:49:39,897 --> 01:49:43,133
YOU CAN SEE IN THE STRUCTURES 

2127
01:49:43,133 --> 01:49:45,102
YOU CAN SEE SIDE CHAINS MAKING 

2128
01:49:45,102 --> 01:49:47,604
IT EASIER TO PRODUCE ATOMIC 

2129
01:49:47,604 --> 01:49:49,206
MODELS FOR THE DENSITY.

2130
01:49:49,206 --> 01:49:54,311
THAT WAS GETTING TO HIGH 

2131
01:49:54,311 --> 01:50:04,521
RESOLUTION AND BECAUSE IT WAS IN

2132
01:50:04,521 --> 01:50:07,057
SITU YOU CAN MAP BACK AND SEE 

2133
01:50:07,057 --> 01:50:09,226
HOW UNIFORM THE DISTRIBUTION IS 

2134
01:50:09,226 --> 01:50:10,427
OR WHETHER THERE'S VARIATIONS 

2135
01:50:10,427 --> 01:50:15,666
FROM THE STRUCTURE.

2136
01:50:15,666 --> 01:50:22,539
ALL OF THAT THIS IS ONE EXAMPLE 

2137
01:50:22,539 --> 01:50:28,479
OF THE TOOLS.

2138
01:50:28,479 --> 01:50:31,381
AND THE BIGGEST CHALLENGE IS HOW

2139
01:50:31,381 --> 01:50:35,285
TO FIND THE FEATURE THAT YOU 

2140
01:50:35,285 --> 01:50:38,255
WANT TO WILL USE TO GET HIGH 

2141
01:50:38,255 --> 01:50:48,465
RESOLUTION.

2142
01:50:52,870 --> 01:50:54,938
AND THESE TOOLS HELP YOU SPEED 

2143
01:50:54,938 --> 01:50:56,373
UP THE PROCESS OF MINING THE 

2144
01:50:56,373 --> 01:50:59,443
DATA AND LOOKING FOR THINGS THAT

2145
01:50:59,443 --> 01:51:00,043
ARE INTERESTING.

2146
01:51:00,043 --> 01:51:09,920
WITHOUT GETTING INTO THE DETAILS

2147
01:51:09,920 --> 01:51:11,421
AND YOU GET THESE THAT REPEAT 

2148
01:51:11,421 --> 01:51:14,057
WITHIN YOUR VOLUMES AND CAN 

2149
01:51:14,057 --> 01:51:16,994
CHOOSE THERE'S SOMETHING I 

2150
01:51:16,994 --> 01:51:18,462
RECOGNIZE I WANT TO GO AFTER 

2151
01:51:18,462 --> 01:51:27,838
THAT AND PUSH IT FORWARD.

2152
01:51:27,838 --> 01:51:28,839
THESE TAKE A MINUTE TO 

2153
01:51:28,839 --> 01:51:33,310
UNDERSTAND HOW THEY LOOK BUT 

2154
01:51:33,310 --> 01:51:35,646
THEY'RE USEFUL.

2155
01:51:35,646 --> 01:51:46,190
WE HAVE THE SARS COV2 VIRUS AND 

2156
01:51:47,791 --> 01:51:50,294
AND YOU ISOLATE WHAT IS ATTACHED

2157
01:51:50,294 --> 01:51:51,862
TO THE MEMBRANE AND IF YOU ZERO 

2158
01:51:51,862 --> 01:51:54,198
IN ON THAT YOU CAN PICK YOUR 

2159
01:51:54,198 --> 01:51:56,466
PARTICLES FROM THE DATA SET AND 

2160
01:51:56,466 --> 01:51:58,368
START PRODUCING STRUCTURES USING

2161
01:51:58,368 --> 01:52:01,138
THE AVERAGING AND IN THIS CASE 

2162
01:52:01,138 --> 01:52:03,740
WE DID CLASSIFICATION AND THAT 

2163
01:52:03,740 --> 01:52:06,210
GIVES US THE STATE OF THE SARS 

2164
01:52:06,210 --> 01:52:08,045
COV2 SPIKE BUT THIS IS AGAIN A 

2165
01:52:08,045 --> 01:52:10,080
GENERAL PURPOSE TOOL YOU CAN USE

2166
01:52:10,080 --> 01:52:17,287
FOR MINING CELLULAR TOMOGRAMS.

2167
01:52:17,287 --> 01:52:19,389
I WANTED TO ACKNOWLEDGE PEOPLE 

2168
01:52:19,389 --> 01:52:29,866
IN MY LAB AND THE TWO MAIN 

2169
01:52:30,367 --> 01:52:34,271
FORCES BEHIND DEVELOPING THIS 

2170
01:52:34,271 --> 01:52:36,540
AND ACKNOWLEDGE COLLABORATORS 

2171
01:52:36,540 --> 01:52:38,375
AND THE SUPPORT THAT MAKES ALL 

2172
01:52:38,375 --> 01:52:38,775
THE WORK POSSIBLE.

2173
01:52:38,775 --> 01:52:48,952
THANK YOU.

2174
01:52:52,389 --> 01:52:54,224
PLEASE  

2175
01:52:54,925 --> 01:53:04,534
>> PLEASE COME UP TO THE
STAGE.

2176
01:53:04,534 --> 01:53:05,569
>> HELLO.

2177
01:53:05,569 --> 01:53:08,905
QUESTION FOR MUNIR AND GNANA 

2178
01:53:08,905 --> 01:53:11,808
COOL WORK WITH THE B CELL 

2179
01:53:11,808 --> 01:53:12,075
RECEPTORS.

2180
01:53:12,075 --> 01:53:20,917
HOW SPECIFIC ON EPITOPE AND 

2181
01:53:20,917 --> 01:53:22,052
PERITOPE AND IT SEEMS IT WOULD 

2182
01:53:22,052 --> 01:53:25,756
AFFECT THE REGION IN SIGNALLING.

2183
01:53:25,756 --> 01:53:31,395
DO YOU THINK DIFFERENT EPITOPES 

2184
01:53:31,395 --> 01:53:32,529
MAY HAVE DIFFERENT SIGNALLING 

2185
01:53:32,529 --> 01:53:34,164
AFFECTS AND IS THAT A BASIS FOR 

2186
01:53:34,164 --> 01:53:36,366
IMMUNODOMINANCE?

2187
01:53:36,366 --> 01:53:41,438
>> I THINK THAT'S WHAT WE
LIKE 

2188
01:53:41,438 --> 01:53:43,473
TO UNDERSTAND RIGHT NOW.

2189
01:53:43,473 --> 01:53:46,443
WHEN WE LOOKED AT THE PURIFIED 

2190
01:53:46,443 --> 01:53:49,413
BCR WHEN WE LOOKED AT HOW THEY 

2191
01:53:49,413 --> 01:53:52,215
BIND THEY LOOK AT THE AFFINITY 

2192
01:53:52,215 --> 01:53:57,220
AND THE RATES ON AND OFF RATES 

2193
01:53:57,220 --> 01:53:58,288
AREN'T VERY DIFFERENT FROM WHAT 

2194
01:53:58,288 --> 01:54:04,594
WE OBSERVED FROM THE SOLUBLE 

2195
01:54:04,594 --> 01:54:06,997
ANTIBODIES AND FRAGMENT.

2196
01:54:06,997 --> 01:54:10,267
THE SPECIFICITY WAS DEFINED BY 

2197
01:54:10,267 --> 01:54:12,035
THE SEQUENCES.

2198
01:54:12,035 --> 01:54:13,704
NOW, WHEN YOU BRING IN THE OTHER

2199
01:54:13,704 --> 01:54:14,805
COMPONENTS OF THE B CELL 

2200
01:54:14,805 --> 01:54:19,076
RECEPTOR AND THAT MIGHT CHANGE 

2201
01:54:19,076 --> 01:54:24,348
BECAUSE WHAT WE THE MODEL BASED 

2202
01:54:24,348 --> 01:54:30,554
ON OUR STRUCTURE AND THE 

2203
01:54:30,554 --> 01:54:33,190
TRANSMISSION AND WE SHOWED 

2204
01:54:33,190 --> 01:54:37,494
THERE'S A LOT OF COMPLEXABILITY 

2205
01:54:37,494 --> 01:54:40,097
AND VARIABILITY AND THE ANTIGEN 

2206
01:54:40,097 --> 01:54:43,600
BINDING WILL AFFECT THAT 

2207
01:54:43,600 --> 01:54:43,900
FLEXIBILITY.

2208
01:54:43,900 --> 01:54:46,837
AND THAT IS WHAT WOULD BE 

2209
01:54:46,837 --> 01:54:49,206
TRANSMITTED TO EACH OF THE 

2210
01:54:49,206 --> 01:54:50,374
DOMAINS OF THE BCR AND 

2211
01:54:50,374 --> 01:54:52,242
SUBSEQUENTLY TO THE SIGNALLING 

2212
01:54:52,242 --> 01:54:54,444
MOLECULE BUT I THINK WE ARE NOW 

2213
01:54:54,444 --> 01:54:57,080
WORKING TO LOOK AT ANTIGEN BOUND

2214
01:54:57,080 --> 01:55:00,283
COMPLEX AND THAT WILL GIVE US 

2215
01:55:00,283 --> 01:55:02,185
THE CLUE TO ANSWER YOU'RE 

2216
01:55:02,185 --> 01:55:02,486
LOOKING FOR.

2217
01:55:02,486 --> 01:55:04,588
>> DO YOU THINK IT MATTERS
IF 

2218
01:55:04,588 --> 01:55:05,822
ONE FAB ISN'T BOUND OR BOTH?

2219
01:55:05,822 --> 01:55:08,558
>> THAT'S AN INTERESTING 

2220
01:55:08,558 --> 01:55:09,593
QUESTION BECAUSE WE THINK BOTH 

2221
01:55:09,593 --> 01:55:12,229
FAB NEEDS TO BE ENGAGED AND WE 

2222
01:55:12,229 --> 01:55:14,464
HAVE A LOT OF DATA LOOKING AT 

2223
01:55:14,464 --> 01:55:21,738
THE SIGNALLING AND IF YOU TOOK A

2224
01:55:21,738 --> 01:55:29,846
MONOMERIC ANTIGEN IN VITRO AS 

2225
01:55:29,846 --> 01:55:30,580
WELL AS EX VIVO B CELLS FROM 

2226
01:55:30,580 --> 01:55:36,620
MOUSE MODELS WE DON'T SEE AC 

2227
01:55:36,620 --> 01:55:38,622
ACTIVATION TO ONE FAB TO ONE 

2228
01:55:38,622 --> 01:55:39,790
ANTIGEN INTERACTION WE THINK 

2229
01:55:39,790 --> 01:55:41,224
BOTH FAB NEED TO BE ENGAGED.

2230
01:55:41,224 --> 01:55:42,959
>> COOL, THANK YOU.

2231
01:55:42,959 --> 01:55:43,260
6:

2232
01:55:43,260 --> 01:55:44,528
>> HI.

2233
01:55:44,528 --> 01:55:48,965
I HAVE A QUESTION FOR ALBERTO.

2234
01:55:48,965 --> 01:55:53,069
FOR YOUR MOLECULAR PATTERN 

2235
01:55:53,069 --> 01:55:54,938
MEANING HAVE YOU CHECKED HOW IT 

2236
01:55:54,938 --> 01:55:58,275
FAIRS ON CELLULAR TOMOGRAMS THAT

2237
01:55:58,275 --> 01:56:06,249
ARE A LITTLE BIT MORE COMPLEX?

2238
01:56:06,249 --> 01:56:08,084
>> HERE I SHOWED THE
EXAMPLE 

2239
01:56:08,084 --> 01:56:10,287
THAT WAS MORE RELEVANT TO THINGS

2240
01:56:10,287 --> 01:56:13,123
HAPPENING IN THE CORE.

2241
01:56:13,123 --> 01:56:19,095
BUT YES, WE CAN COMBINE 

2242
01:56:19,095 --> 01:56:21,164
INTERIORS AND WE HAVE SEVERAL 

2243
01:56:21,164 --> 01:56:22,866
CASES WE VALIDATED THESE AND 

2244
01:56:22,866 --> 01:56:24,000
HOPEFULLY AN UPCOMING 

2245
01:56:24,000 --> 01:56:24,901
PUBLICATION WILL HAVE THE 

2246
01:56:24,901 --> 01:56:25,902
DETAILS BUT THE ANSWER IS YES.

2247
01:56:25,902 --> 01:56:27,471
>> IT'S A FASCINATING
TOOL.

2248
01:56:27,471 --> 01:56:31,174
I'M REALLY EXCITED.

2249
01:56:31,174 --> 01:56:32,843
>> FOR US IT'S BEEN A GAME

2250
01:56:32,843 --> 01:56:37,581
CHANGER BAYS NOW WE DON'T NEED 

2251
01:56:37,581 --> 01:56:41,818
TO SPEND TIME IN THE DATA.

2252
01:56:41,818 --> 01:56:46,122
>> THIS MAY BE A QUESTION 

2253
01:56:46,122 --> 01:56:47,224
PRINCIPALLY FOR RICK IT'S BEEN 

2254
01:56:47,224 --> 01:56:50,293
GREAT TO HAVE ENVELOPE PEOPLE IN

2255
01:56:50,293 --> 01:56:51,194
THE MEETING.

2256
01:56:51,194 --> 01:56:52,963
I WAS TAKEN BY THE FACT YOU 

2257
01:56:52,963 --> 01:56:56,533
THOUGHT THE TILT OF THE TRANS 

2258
01:56:56,533 --> 01:56:58,201
MEMBRANE DOMAIN CHANGED ON 

2259
01:56:58,201 --> 01:56:59,669
BINDING AND THAT WOULD 

2260
01:56:59,669 --> 01:57:00,604
PROBABLY -- IT'S ALWAYS 

2261
01:57:00,604 --> 01:57:04,574
INTERESTING TO THINK OF CAUSE 

2262
01:57:04,574 --> 01:57:05,775
AND EFFECT BUT YOU POINTED OUT 

2263
01:57:05,775 --> 01:57:09,346
IT WOULD CHANGE THE LIPID 

2264
01:57:09,346 --> 01:57:11,214
FLUIDITY AND WONDER IF YOU CAN 

2265
01:57:11,214 --> 01:57:14,484
DRIVE IT IN THE OPPOSITE 

2266
01:57:14,484 --> 01:57:14,751
DIRECTION.

2267
01:57:14,751 --> 01:57:18,021
CAN YOU CHANGE THE AFFINITY FOR 

2268
01:57:18,021 --> 01:57:20,023
LIGAND BY CHANGING THE LIPID 

2269
01:57:20,023 --> 01:57:28,064
COMPOSITION OF THE MEMBRANES?

2270
01:57:28,064 --> 01:57:30,901
>> ONE QUESTION WE ASKED
IS THE 

2271
01:57:30,901 --> 01:57:34,104
BCR WORK WAS DONE WITH 

2272
01:57:34,104 --> 01:57:34,504
DETERGENTS.

2273
01:57:34,504 --> 01:57:37,340
WE WANTED TO SEE WHAT HAPPENS 

2274
01:57:37,340 --> 01:57:42,078
WHEN HAVE YOU THE SIMPLE PRPC 

2275
01:57:42,078 --> 01:57:43,280
VERSUS LIKE A B CELL MEMBRANE 

2276
01:57:43,280 --> 01:57:45,382
AND WE DO SEE A DIFFERENCE.

2277
01:57:45,382 --> 01:57:50,186
I THINK THE MEMBRANE MIGHT 

2278
01:57:50,186 --> 01:57:52,622
COMPOSITION YOU'RE LOOKING AT 

2279
01:57:52,622 --> 01:58:00,297
THE BCRs MAY AFFECT THE WAY 

2280
01:58:00,297 --> 01:58:01,364
THISE THE INFORMATION GETS BOUND

2281
01:58:01,364 --> 01:58:05,502
TO THE TRANS MEMBRANE REGIONS.

2282
01:58:05,502 --> 01:58:11,541
SO I THINK ONE OF THE MAIN THING

2283
01:58:11,541 --> 01:58:13,443
WE SEE IS WHENEVER YOU HAVE THE 

2284
01:58:13,443 --> 01:58:16,546
MAME BRAIN THE MOTIONS ARE 

2285
01:58:16,546 --> 01:58:17,747
LARGER.

2286
01:58:17,747 --> 01:58:20,016
AND SO WE ARE STILL TRYING TO 

2287
01:58:20,016 --> 01:58:24,955
HAVE ASSIMILATION AND IT'S NOT 

2288
01:58:24,955 --> 01:58:26,623
BY CHANCE YOU'RE SEEING 

2289
01:58:26,623 --> 01:58:26,923
DIFFERENCES.

2290
01:58:26,923 --> 01:58:27,691
WE'RE ELEVATING THAT PART RIGHT 

2291
01:58:27,691 --> 01:58:29,326
NOW.

2292
01:58:29,326 --> 01:58:33,029
>> THANK YOU.

2293
01:58:33,029 --> 01:58:34,341
[APPLAUSE]

2294
01:58:45,102 --> 01:58:47,571
IT'S A GREAT PLEASURE FOR ME TO 

2295
01:58:47,571 --> 01:58:52,543
INTRODUCE OUR CENTER THAT IS 

2296
01:58:52,543 --> 01:58:55,713
DIRECTED BY TATIANA POLENOVA AND

2297
01:58:55,713 --> 01:58:57,581
MYSELF AND I'LL GO VERY BRIEFLY 

2298
01:58:57,581 --> 01:58:58,549
THROUGH A BIT OF THE BACKGROUND 

2299
01:58:58,549 --> 01:59:08,793
OF THE CENTER.

2300
01:59:13,864 --> 01:59:19,570
WE HAVE P.I.s ALL OVER THE WORLD

2301
01:59:19,570 --> 01:59:30,080
AND WHAT IS IMPORTANT WE HAVE 

2302
01:59:43,727 --> 01:59:52,203
WORKED ON NUCLEAR INTERPMPORT A 

2303
01:59:52,203 --> 01:59:54,004
INTEGRATION AND WE HAVE CORES 

2304
01:59:54,004 --> 01:59:56,907
AND COLLABORATORS AND THE 

2305
01:59:56,907 --> 02:00:00,010
COMPUTATIONAL CORE AND CRYO EM 

2306
02:00:00,010 --> 02:00:05,983
CORE AND VIROLOGY CORE.

2307
02:00:05,983 --> 02:00:16,527
IN THE DEVELOPMENT CORE AND KWAE

2308
02:00:18,696 --> 02:00:19,630
HAVE A MENTORED SCIENTIST 

2309
02:00:19,630 --> 02:00:21,999
PROGRAM AND STARTED A RESEARCH 

2310
02:00:21,999 --> 02:00:27,304
EMBEDDING PROGRAM WHICH IS 

2311
02:00:27,304 --> 02:00:29,607
UNFORTUNATELY SORT OF THROWING A

2312
02:00:29,607 --> 02:00:31,609
WRENCH THAT THAT AND HAVE 

2313
02:00:31,609 --> 02:00:33,677
SEVERAL EXAMPLES WHERE PEOPLE 

2314
02:00:33,677 --> 02:00:35,446
FROM ONE P.I.'S LAB GO TO 

2315
02:00:35,446 --> 02:00:37,548
ANOTHER P.I.s LAB AND STAY IN 

2316
02:00:37,548 --> 02:00:39,783
THAT LAB FOR WEEKS OR MONTHS AND

2317
02:00:39,783 --> 02:00:41,552
LEARN TECHNOLOGIES AND THEN 

2318
02:00:41,552 --> 02:00:43,287
BRING THAT BACK TO THEIR HOME 

2319
02:00:43,287 --> 02:00:45,990
LAB AND THIS IS SOMETHING THAT 

2320
02:00:45,990 --> 02:00:48,926
WE STRONGLY FEEL IS A BIG 

2321
02:00:48,926 --> 02:00:50,728
ADVANCEMENT IN TRYING TO TRAIN 

2322
02:00:50,728 --> 02:00:58,502
OUR NEXT GENERATION SCIENTISTS.

2323
02:00:58,502 --> 02:01:00,404
 OUR CENTER ALWAYS HAD A STRONG 

2324
02:01:00,404 --> 02:01:01,305
EMPHASIS ON TECHNOLOGY 

2325
02:01:01,305 --> 02:01:03,107
DEVELOPMENT AND ALL OF OUR CORES

2326
02:01:03,107 --> 02:01:04,575
CONTRIBUTE TO ALL THE PROJECT 

2327
02:01:04,575 --> 02:01:06,644
BUT WE ALSO REALLY THINK ABOUT 

2328
02:01:06,644 --> 02:01:11,882
WHERE WE COULD PUSH THE 

2329
02:01:11,882 --> 02:01:13,584
TECHNOLOGIES.

2330
02:01:13,584 --> 02:01:16,487
AND UP PARTICULAR THE MOLECULE 

2331
02:01:16,487 --> 02:01:19,089
CORRELATED MICROSCOPY CORE 

2332
02:01:19,089 --> 02:01:20,424
CONTRIBUTES TO THE DIFFERENT 

2333
02:01:20,424 --> 02:01:22,726
PROJECTS IN THE LAB AND THE 

2334
02:01:22,726 --> 02:01:23,694
COMPUTATIONAL CORE.

2335
02:01:23,694 --> 02:01:27,264
WE HAVE PUBLISHED OVER THE LAST 

2336
02:01:27,264 --> 02:01:29,233
YEAR AND SEVERAL OF THE 

2337
02:01:29,233 --> 02:01:34,572
PUBLICATIONS ARE PUBLICATIONS 

2338
02:01:34,572 --> 02:01:40,844
BETWEEN DIFFERENT CENTERS 

2339
02:01:40,844 --> 02:01:44,148
UNFORTUNATELY MOST OUR POSTERS 

2340
02:01:44,148 --> 02:01:45,983
WERE YESTERDAY AND HOPEFULLY YOU

2341
02:01:45,983 --> 02:01:47,751
CAN SEE THE POST DOCS LISTED 

2342
02:01:47,751 --> 02:01:51,689
HERE AND I'M SURE THEY'LL TALK 

2343
02:01:51,689 --> 02:01:52,957
TO YOU AGAIN AND THERE ARE FOUR 

2344
02:01:52,957 --> 02:01:55,059
POSTERS WE HAVE TODAY.

2345
02:01:55,059 --> 02:01:59,296
THE TALKS TODAY ARE FROM THREE 

2346
02:01:59,296 --> 02:02:02,399
PEOPLE TATYANA WILL TALK ABOUT 

2347
02:02:02,399 --> 02:02:07,738
THE MATURATION INHIBITOR BINDING

2348
02:02:07,738 --> 02:02:09,974
AND SHE WILL HIGHLIGHT THE 

2349
02:02:09,974 --> 02:02:14,778
TECHNOLOGY OF SPINNING SOLID 

2350
02:02:14,778 --> 02:02:19,617
STATE AND MEL IS LA KANE WILL 

2351
02:02:19,617 --> 02:02:22,586
TALK ABOUT THE RANBP2 IN HIV-1 

2352
02:02:22,586 --> 02:02:23,988
INFECTION AND YONG WILL TALK 

2353
02:02:23,988 --> 02:02:27,691
ABOUT THE BIOLOGY OF HOST VIRUS 

2354
02:02:27,691 --> 02:02:28,025
INTERACTIONS.

2355
02:02:28,025 --> 02:02:29,994
WITH THIS I HAND IT OVER TO 

2356
02:02:29,994 --> 02:02:33,998
TATIANA WHO WILL TALK ABOUT THE 

2357
02:02:33,998 --> 02:02:43,307
SCIENCE THAT'S GOING ON.

2358
02:02:43,307 --> 02:02:45,909
>> GOOD MORNING,
EVERYBODY.

2359
02:02:45,909 --> 02:02:46,977
IT'S ALWAYS GREAT TO BE HERE AND

2360
02:02:46,977 --> 02:02:49,046
THANK YOU FOR BRINGING US TO THE

2361
02:02:49,046 --> 02:02:50,147
SCIENCE CELEBRATION.

2362
02:02:50,147 --> 02:02:53,817
AND I KNOW THAT WE ARE 

2363
02:02:53,817 --> 02:02:54,518
INTEGRATING THAT WITH THE 

2364
02:02:54,518 --> 02:02:55,819
TEMPERATURE IN THE ROOM SO I'LL 

2365
02:02:55,819 --> 02:02:57,788
TRY TO BE ENERGETIC TO GIVE YOU 

2366
02:02:57,788 --> 02:03:01,058
AN OVERVIEW OF WHAT WE DO AND 

2367
02:03:01,058 --> 02:03:02,393
IT'S SOMETHING THAT HAS NOT YET 

2368
02:03:02,393 --> 02:03:06,797
BEEN DISCUSSED OVER THE PAST DAY

2369
02:03:06,797 --> 02:03:11,702
AND A HALF, MATURATION WITH THE 

2370
02:03:11,702 --> 02:03:13,671
VIRAL HIV LIFE CYCLE TARGETED BY

2371
02:03:13,671 --> 02:03:15,105
SMALL MOLECULES.

2372
02:03:15,105 --> 02:03:17,975
WHAT I WILL TELL YOU TODAY ABOUT

2373
02:03:17,975 --> 02:03:20,144
IS THE WORK TRULY AN INTEGRATION

2374
02:03:20,144 --> 02:03:21,345
FROM SEVERAL CENTERS AND THE 

2375
02:03:21,345 --> 02:03:25,149
LONG STANDING COLLABORATION 

2376
02:03:25,149 --> 02:03:35,693
BETWEEN THE CORE AND MYSELF AND 

2377
02:03:36,026 --> 02:03:39,797
THE POSTDOC CENTER LABS DONE BY 

2378
02:03:39,797 --> 02:03:43,967
TALENTED PEOPLE AT THE 

2379
02:03:43,967 --> 02:03:54,511
UNIVERSITY OF DELAWARE AND IS WE

2380
02:04:00,818 --> 02:04:05,756
HAVE AN NMR CORE CALLED SPINNING

2381
02:04:05,756 --> 02:04:06,757
NMR TO LOOK AT STRUCTURAL 

2382
02:04:06,757 --> 02:04:16,900
BIOLOGY.

2383
02:04:21,305 --> 02:04:22,272
THEY TALKED ABOUT THE DIFFERENT 

2384
02:04:22,272 --> 02:04:27,111
TYPE OF NMR AND THIS IS MAGIC 

2385
02:04:27,111 --> 02:04:28,178
ANGLE SPINNING NMR.

2386
02:04:28,178 --> 02:04:29,279
WHAT'S SO MAGIC?

2387
02:04:29,279 --> 02:04:31,515
AS FAR AS THE APPLICATIONS GO WE

2388
02:04:31,515 --> 02:04:34,651
CAN LOOK AT THINGS THAT ARE 

2389
02:04:34,651 --> 02:04:40,691
PRETTY LARGE AND INTACT CELLS 

2390
02:04:40,691 --> 02:04:45,929
AND ORGANELLES AND THERE'S CRYO 

2391
02:04:45,929 --> 02:04:48,866
ELECTRIC MICROSCOPY AND ISOLATE 

2392
02:04:48,866 --> 02:04:51,235
YOUR ASSEMBLIES OR OR PARTICLES 

2393
02:04:51,235 --> 02:04:53,437
AND FOR MAGIC ANGLE SPINNING AND

2394
02:04:53,437 --> 02:04:56,039
MORE WE USE A SPECIAL SAMPLE 

2395
02:04:56,039 --> 02:04:58,509
CONTAINER CALLED ROTORS.

2396
02:04:58,509 --> 02:05:01,979
WE HAVE TO ROTATE THE SAMPLES 

2397
02:05:01,979 --> 02:05:05,215
VERY FAST UP TO 110,000 

2398
02:05:05,215 --> 02:05:09,753
REVOLUTIONS PER SECOND AT TIME 

2399
02:05:09,753 --> 02:05:12,055
110 KILO-HERTZ AND THIS IS 

2400
02:05:12,055 --> 02:05:15,025
IMPORTANT FOR THE TYPE OF 

2401
02:05:15,025 --> 02:05:15,526
APPLICATIONS I WILL BE 

2402
02:05:15,526 --> 02:05:21,632
DISCUSSING.

2403
02:05:21,632 --> 02:05:27,437
WE HAVE MULTIDIMENSIONAL 

2404
02:05:27,437 --> 02:05:29,640
EXPERIMENTS AND WE CAN DETERMINE

2405
02:05:29,640 --> 02:05:30,841
STATIC STRUCTURES AND WE 

2406
02:05:30,841 --> 02:05:32,309
DEMONSTRATED OVER THE YEARS AND 

2407
02:05:32,309 --> 02:05:34,511
ALSO WHAT'S IMPORTANT IS WE GET 

2408
02:05:34,511 --> 02:05:36,914
INFORMATION ABOUT DYNAMICS FROM 

2409
02:05:36,914 --> 02:05:37,347
THE MEASUREMENTS.

2410
02:05:37,347 --> 02:05:40,584
AND WHEN WE COUPLE OUR 

2411
02:05:40,584 --> 02:05:41,819
EXPERIMENTS WITH POWERFUL 

2412
02:05:41,819 --> 02:05:45,856
MOLECULAR DYNAMIC SIMULATIONS 

2413
02:05:45,856 --> 02:05:49,326
AND MECHANICAL STIMULATIONS WE 

2414
02:05:49,326 --> 02:05:51,695
CAN LOOK AT DYNAMIC STRUCTURES.

2415
02:05:51,695 --> 02:05:55,365
LOOKING AT THE ARCHITECTURE AND 

2416
02:05:55,365 --> 02:05:55,599
MOTION.

2417
02:05:55,599 --> 02:05:59,169
THE ADVANTAGE IS QUITE A FEW.

2418
02:05:59,169 --> 02:06:02,906
AND IN SOLUTION NMR WE CAN SEE 

2419
02:06:02,906 --> 02:06:05,943
THE QUOTE, UNQUOTE, IN VISIBLE 

2420
02:06:05,943 --> 02:06:07,244
STRUCTURES AND I'LL SHOW SOME 

2421
02:06:07,244 --> 02:06:11,582
EXAMPLES TODAY AND WE CAN LOOK 

2422
02:06:11,582 --> 02:06:13,984
AT SMALL MOLECULES WITH DETAIL 

2423
02:06:13,984 --> 02:06:19,723
AND THERE'S NO SYMMETRY REQUIRED

2424
02:06:19,723 --> 02:06:21,925
AND AS THE ONLY EXPERIMENTAL 

2425
02:06:21,925 --> 02:06:24,628
TECHNIQUE THAT YIELDS DYNAMIC 

2426
02:06:24,628 --> 02:06:27,698
INFORMATION IN ASSEMBLIES WITH 

2427
02:06:27,698 --> 02:06:28,665
RESOLUTION ON A BROAD 

2428
02:06:28,665 --> 02:06:30,667
TEMPERATURE RANGE FROM 

2429
02:06:30,667 --> 02:06:33,971
PHYSIOLOGICAL TO CRYO GENIC.

2430
02:06:33,971 --> 02:06:35,038
OF COURSE THERE ARE SOME 

2431
02:06:35,038 --> 02:06:37,841
COMPLICATIONS BUT TODAY I WANT 

2432
02:06:37,841 --> 02:06:39,910
TO TALK ABOUT AND HIGHLIGHT THE 

2433
02:06:39,910 --> 02:06:43,180
POWER OF LOOKING AT HIV 

2434
02:06:43,180 --> 02:06:43,914
STRUCTURAL VIRAL GENE.

2435
02:06:43,914 --> 02:06:46,850
WE'RE INTERESTED IN A NUMBER OF 

2436
02:06:46,850 --> 02:06:50,721
STEPS IN THE VIRAL LIFE CYCLE.

2437
02:06:50,721 --> 02:06:52,923
AND WE HAVE LOOKED AT THE GAPS 

2438
02:06:52,923 --> 02:06:55,692
IN ARCHITECTURE AND LOOKING AT 

2439
02:06:55,692 --> 02:06:56,693
THE INTERACTIONS WITH 

2440
02:06:56,693 --> 02:06:57,594
MICROTUBULES AND THERE'S A 

2441
02:06:57,594 --> 02:06:59,730
POSTER AND WE HAVE LOOKED AT 

2442
02:06:59,730 --> 02:07:00,898
INTERACTIONS WITH THE FACTOR AND

2443
02:07:00,898 --> 02:07:03,834
AS I TOLD YOU WE'LL FOCUS ON THE

2444
02:07:03,834 --> 02:07:04,768
MATURATION PART OF THE VIRAL 

2445
02:07:04,768 --> 02:07:14,978
LIFE CYCLE.

2446
02:07:16,380 --> 02:07:21,985
AND THIS IS A PROCESS TIGHTLY 

2447
02:07:21,985 --> 02:07:25,255
REGULATED SPATIALLY AND THE LAST

2448
02:07:25,255 --> 02:07:32,129
PART IS THE PEPTIDE AND THE 

2449
02:07:32,129 --> 02:07:34,364
MATURE VARIANTS WITH THE 

2450
02:07:34,364 --> 02:07:36,166
CANONICAL CAPSIDS AND THE LAST 

2451
02:07:36,166 --> 02:07:39,670
STEP IS ACCOMPANIED BY A LARGE 

2452
02:07:39,670 --> 02:07:42,639
MORPHOLOGICAL CHANGE FROM 

2453
02:07:42,639 --> 02:07:45,976
IMMATURE TO MATURE LATTICE AND 

2454
02:07:45,976 --> 02:07:50,447
THE ORGANIZATIONAL WILL DCA IS 

2455
02:07:50,447 --> 02:07:52,649
SUCH THE CR TERMINAL DOMAIN HAS 

2456
02:07:52,649 --> 02:08:00,691
THE SHAPE OF A GOBLET AND SO 

2457
02:08:00,691 --> 02:08:01,959
MATURATION HAS BECOME AN 

2458
02:08:01,959 --> 02:08:03,560
ATTRACTION FOR THE INHIBITORS 

2459
02:08:03,560 --> 02:08:05,228
AND OTHER COLLEAGUES HAVE WORKED

2460
02:08:05,228 --> 02:08:08,198
OVER THE YEARS ON DESIGNING 

2461
02:08:08,198 --> 02:08:09,566
POTENT INHIBIT.

2462
02:08:09,566 --> 02:08:13,971
THE FIRST IS THE MOLECULE WHICH 

2463
02:08:13,971 --> 02:08:19,176
IS WILL AN ACID AND THERE'S ALSO

2464
02:08:19,176 --> 02:08:21,845
A UNIQUE INHIBITOR DESIGN BY 

2465
02:08:21,845 --> 02:08:22,646
PFIZER.

2466
02:08:22,646 --> 02:08:24,514
WHAT HAPPENS WHEN THE MATURATION

2467
02:08:24,514 --> 02:08:28,652
INHIBITORS ARE ADDED TO THE 

2468
02:08:28,652 --> 02:08:38,829
VIRUS THEIR VIRIONS ARE FORMED 

2469
02:08:38,829 --> 02:08:40,664
AND STABILIZE THE BUNDLE.

2470
02:08:40,664 --> 02:08:42,933
THE PROCESS HAS BEEN BEAUTIFULLY

2471
02:08:42,933 --> 02:08:45,635
STUDIED BY MANY PEOPLE IN THE 

2472
02:08:45,635 --> 02:08:47,804
AUDIENCE EXPERIMENTS IN 

2473
02:08:47,804 --> 02:08:50,640
CRYSTALLOGRAPHY AND 

2474
02:08:50,640 --> 02:08:52,476
COMPUTATIONALLY AND IT MICRO-ID 

2475
02:08:52,476 --> 02:08:55,679
THERE'S BEAUTIFUL INSIGHTS AND 

2476
02:08:55,679 --> 02:08:57,481
DECIDED TO LOOK INTO THE 

2477
02:08:57,481 --> 02:08:59,516
COLLABORATION A FEW YEARS AGO 

2478
02:08:59,516 --> 02:09:03,186
BECAUSE ALL OF THESE STUDIES AT 

2479
02:09:03,186 --> 02:09:05,822
THE TIME HAD DETAILS OF THE 

2480
02:09:05,822 --> 02:09:06,790
BINDING WERE NOT CLEAR.

2481
02:09:06,790 --> 02:09:11,561
SO WE CONTRIBUTED TO THE 

2482
02:09:11,561 --> 02:09:14,064
PLETHORA OF EXPERIMENTS AND 

2483
02:09:14,064 --> 02:09:15,766
CALCULATIONS BY SOLVING THE 

2484
02:09:15,766 --> 02:09:17,968
STRUCTURAL DVM PUBLISHED LAST 

2485
02:09:17,968 --> 02:09:19,269
YEAR AND QUICKLY WALK YOU 

2486
02:09:19,269 --> 02:09:20,504
THROUGH THAT BEFORE I TALK SOME 

2487
02:09:20,504 --> 02:09:21,738
OF THE RECENT RESULTS BECAUSE 

2488
02:09:21,738 --> 02:09:32,215
I'M NOT SURE I DISCUSSED IT.

2489
02:09:52,903 --> 02:09:57,507
WE HAVE THE PROTONS AN 

2490
02:09:57,507 --> 02:09:59,342
DETERMINED THE PROTEIN STRUCTURE

2491
02:09:59,342 --> 02:10:08,018
AND USED PROTONS AND LOOKED AT 

2492
02:10:08,018 --> 02:10:11,354
INHIBITOR INTERACTIONS AND ALSO 

2493
02:10:11,354 --> 02:10:15,892
AS MANY KNOW FROM THE SAMPLING 

2494
02:10:15,892 --> 02:10:19,930
OF THE VIRUS THIS WAS REQUIRED 

2495
02:10:19,930 --> 02:10:23,500
AND IT'S EXPLOIT THIS PROPERTY 

2496
02:10:23,500 --> 02:10:25,168
TO IN VITRO ASSEMBLIES AND 

2497
02:10:25,168 --> 02:10:30,707
LOOKING AT THE IP6 DYNAMICS BY 

2498
02:10:30,707 --> 02:10:32,442
USING PHOSPHORUS NMR SO THIS IS 

2499
02:10:32,442 --> 02:10:33,677
OUR TOOL KIT.

2500
02:10:33,677 --> 02:10:34,878
LET'S LOOK AT HOW WE LOOKED AT 

2501
02:10:34,878 --> 02:10:45,122
THE ASSEMBLY.

2502
02:11:06,743 --> 02:11:10,680
AND THE RESOLUTION IS SO HIGH WE

2503
02:11:10,680 --> 02:11:14,951
CAN PULL OUT DISTANCE RESTRAINTS

2504
02:11:14,951 --> 02:11:18,522
AND THEY'RE SOME OF THE BEST 

2505
02:11:18,522 --> 02:11:21,324
DEFINED STRUCTURE AND WE CAN SEE

2506
02:11:21,324 --> 02:11:24,327
DYNAMICALLY DISORDERED PARTS OF 

2507
02:11:24,327 --> 02:11:26,463
THE PROTEIN NOT VISIBLE IN THE 

2508
02:11:26,463 --> 02:11:30,800
CRYO EM AND DETERMINED BY THE 

2509
02:11:30,800 --> 02:11:34,604
RESIDUES AS WELL AS THE FIRST OF

2510
02:11:34,604 --> 02:11:35,739
THE CTD.

2511
02:11:35,739 --> 02:11:39,609
HOW DO WE LOOK AT THE BINDING OF

2512
02:11:39,609 --> 02:11:39,876
BVM?

2513
02:11:39,876 --> 02:11:41,545
WE HAVE SPECIAL CORRELATIONS 

2514
02:11:41,545 --> 02:11:43,213
BETWEEN THE PROTEINS AND THE 

2515
02:11:43,213 --> 02:11:44,381
SMALL MOLECULES AND YOU CAN SEE 

2516
02:11:44,381 --> 02:11:52,689
HERE THE OVERLAYS OF THE ASIMPLY

2517
02:11:52,689 --> 02:11:56,193
ASSEMBLY AND CONCLUDED IT'S 

2518
02:11:56,193 --> 02:12:00,597
BOUND WITH IP6 ALSO WHAT OTHERS 

2519
02:12:00,597 --> 02:12:01,965
DISCOVERED AND IMPORTANTLY WE 

2520
02:12:01,965 --> 02:12:09,506
CAN UNAMBIGUOUSLY DETERMINE THE 

2521
02:12:09,506 --> 02:12:16,680
MOLECULE AND IT POINTS TO THE 6 

2522
02:12:16,680 --> 02:12:22,652
AND POINTS TO THIS REGION AN 

2523
02:12:22,652 --> 02:12:24,221
I'LL TELL YOU BASED ON MANY 

2524
02:12:24,221 --> 02:12:25,956
EXPERIMENTS AND THE CALCULATION 

2525
02:12:25,956 --> 02:12:32,762
OF THE STRUCTURE WE FOUND BVM 

2526
02:12:32,762 --> 02:12:40,670
BINDS SIMULTANEOUS LY WE SEE IT 

2527
02:12:40,670 --> 02:12:51,214
BINDS TO THREE CHAINS AND THAT'S

2528
02:12:51,881 --> 02:12:55,352
WHY YOU KNOW IT DIDN'T DEVELOP 

2529
02:12:55,352 --> 02:12:57,988
TO CLINICAL TRIALS AND WE 

2530
02:12:57,988 --> 02:13:01,124
CHARACTERIZED THE DETAILS IN THE

2531
02:13:01,124 --> 02:13:04,728
TWO THAT ARE INTERESTING AND 

2532
02:13:04,728 --> 02:13:07,130
IMPORTANT THE MUTATION THE 

2533
02:13:07,130 --> 02:13:10,000
VARIANTS WE HAVE STUDIED AND 

2534
02:13:10,000 --> 02:13:12,669
BASED ON EXPERIMENTAL DATA AND 

2535
02:13:12,669 --> 02:13:14,638
MORE MICROSCOPY BASED WE 

2536
02:13:14,638 --> 02:13:15,505
CONCLUDED SEVERAL INTERESTING 

2537
02:13:15,505 --> 02:13:16,506
THINGS.

2538
02:13:16,506 --> 02:13:19,709
FIRST IF WE LOOK AT THE MUTANT 

2539
02:13:19,709 --> 02:13:21,811
WHAT HAPPENS IS BVM SIMPLY DOES 

2540
02:13:21,811 --> 02:13:22,712
NOT BYPRODUCT THERE AND THE 

2541
02:13:22,712 --> 02:13:24,814
VARIANT IS MORE DYNAMIC.

2542
02:13:24,814 --> 02:13:28,752
WE CAN GET THIS INFORMATION 

2543
02:13:28,752 --> 02:13:32,088
DIRECTLY FROM THE NMR SPECTRA 

2544
02:13:32,088 --> 02:13:39,663
AND THIS DOES BIND BVM BUT MORE 

2545
02:13:39,663 --> 02:13:40,630
VARIANT THAN THE WILD TYPE AND 

2546
02:13:40,630 --> 02:13:43,099
IT'S MORE DYNAMIC THAN THE WILD 

2547
02:13:43,099 --> 02:13:43,400
TYPE.

2548
02:13:43,400 --> 02:13:49,973
THIS IS A VERY UNIQUE WAY THE 

2549
02:13:49,973 --> 02:13:50,707
RESISTANCE IS CONFERRED.

2550
02:13:50,707 --> 02:13:54,044
ANOTHER WAY TO LOOK AT WILD TYPE

2551
02:13:54,044 --> 02:13:56,146
AND RESISTANCE VARIANTS IS TO 

2552
02:13:56,146 --> 02:13:59,249
LOOK AT THE PHOSPHORUS BY 

2553
02:13:59,249 --> 02:14:00,717
PROBING IP6 ITSELF AND 

2554
02:14:00,717 --> 02:14:03,787
ORIENTATION AND DYNAMICS AND 

2555
02:14:03,787 --> 02:14:05,388
WALK YOU THROUGH THAT QUICKLY 

2556
02:14:05,388 --> 02:14:06,990
BECAUSE IT'S INTERESTING.

2557
02:14:06,990 --> 02:14:09,426
IF WE DO THE EXPERIMENTS WE'RE 

2558
02:14:09,426 --> 02:14:13,563
LOOKING AT THE SIGNALS AND ONE 

2559
02:14:13,563 --> 02:14:19,202
PROBES BOTH DYNAMIC MOLECULES 

2560
02:14:19,202 --> 02:14:22,439
AND HERE WILD TYPE AND THEN A1V 

2561
02:14:22,439 --> 02:14:25,642
VARIANTS WITH OR WITHOUT BVM OR 

2562
02:14:25,642 --> 02:14:27,477
THE SIGNAL AND THAT'S A GREAT 

2563
02:14:27,477 --> 02:14:30,680
CONTROL EXPERIMENT.

2564
02:14:30,680 --> 02:14:32,982
NOW WE HAVE CROSS 

2565
02:14:32,982 --> 02:14:34,718
PHOSPHORYLIZATION AND THERE IS 

2566
02:14:34,718 --> 02:14:35,919
SOMETHING INTERESTING HERE.

2567
02:14:35,919 --> 02:14:39,789
WHEN WE HAVE NO B VM THERE IS NO

2568
02:14:39,789 --> 02:14:40,023
SIGNAL.

2569
02:14:40,023 --> 02:14:42,926
THIS TELLS US IP6 IS DYNAMIC 

2570
02:14:42,926 --> 02:14:45,462
INSIDE THE CORE ON THE TIME 

2571
02:14:45,462 --> 02:14:48,098
SCALES OF MILLISECONDS.

2572
02:14:48,098 --> 02:14:51,234
IT MEANS WE KNOW FROM ALL THE 

2573
02:14:51,234 --> 02:14:52,736
EXPERIMENTS THIS ROTATION INSIDE

2574
02:14:52,736 --> 02:14:53,970
THE CORE.

2575
02:14:53,970 --> 02:14:56,940
SO NOW WHAT HAPPENS WHEN WE ADD 

2576
02:14:56,940 --> 02:14:58,274
BVM?

2577
02:14:58,274 --> 02:15:00,310
IN THE CASE OF THE WILD TYPE WE 

2578
02:15:00,310 --> 02:15:01,745
GET SIGNAL BUT YOU CAN SEE THERE

2579
02:15:01,745 --> 02:15:03,880
ARE DIFFERENT TYPES OF SIGNALS.

2580
02:15:03,880 --> 02:15:07,517
SO A1V THERE IS NO SIGNAL WHICH 

2581
02:15:07,517 --> 02:15:10,854
ARE IS NOT SURPRISING BECAUSE 

2582
02:15:10,854 --> 02:15:13,656
A1V DOES NOT BIND BVM AND IT'S 

2583
02:15:13,656 --> 02:15:13,990
DYNAMIC.

2584
02:15:13,990 --> 02:15:20,230
THE EXPERIMENTS ARE QUICK.

2585
02:15:20,230 --> 02:15:23,733
THIS COULD BE A NICE SCREENING.

2586
02:15:23,733 --> 02:15:25,769
WE CAN DO CORRELATION 

2587
02:15:25,769 --> 02:15:27,670
EXPERIMENTS IN TWO DIMENSIONAL 

2588
02:15:27,670 --> 02:15:30,006
WAYS AND THESE ARE CORRELATIONS 

2589
02:15:30,006 --> 02:15:32,275
BETWEEN PHOSPHORUS AND PROTON 

2590
02:15:32,275 --> 02:15:33,610
ATOMS OF THE PROTEIN AND BASED 

2591
02:15:33,610 --> 02:15:36,513
ON THE EXPERIMENT AND YOU CAN 

2592
02:15:36,513 --> 02:15:39,082
SEE THE WILD TYPE IN BLACK AND A

2593
02:15:39,082 --> 02:15:40,350
VARIANT IN PURPLE.

2594
02:15:40,350 --> 02:15:43,086
VERY DIFFERENT CORRELATIONS.

2595
02:15:43,086 --> 02:15:46,423
THIS TELLS US IP6 ADOPTS A 

2596
02:15:46,423 --> 02:15:49,726
DIFFERENT ORIENTATION AND YOU 

2597
02:15:49,726 --> 02:15:52,729
CAN SEE IN THE WILD TYPE IT'S 

2598
02:15:52,729 --> 02:15:54,597
TILTED IN THIS WAY AND IT ADOPTS

2599
02:15:54,597 --> 02:15:59,169
A MORE PARALLEL ORIENTATION.

2600
02:15:59,169 --> 02:16:00,670
THESE ARE THE EXPERIMENTS WE 

2601
02:16:00,670 --> 02:16:02,939
APPLY WHEN WE LOOK AT THE 

2602
02:16:02,939 --> 02:16:03,540
MOLECULE.

2603
02:16:03,540 --> 02:16:04,541
THIS IS THE PFIZER COMPOUND WITH

2604
02:16:04,541 --> 02:16:09,579
A PROPELLER SHAPE AND THREE 

2605
02:16:09,579 --> 02:16:16,719
CHEMICAL GROUPS AND BENZENE AND 

2606
02:16:16,719 --> 02:16:27,263
TRI FLUORO ANTIMER AND WE SOLVED

2607
02:16:34,137 --> 02:16:36,739
THE SOLVED OF THE CRYSTAL AND 

2608
02:16:36,739 --> 02:16:41,678
THEN LOOKED AT THE BINDING.

2609
02:16:41,678 --> 02:16:47,317
THE FIRST THING DISCOVERED WAS 

2610
02:16:47,317 --> 02:16:49,986
INTERESTING.

2611
02:16:49,986 --> 02:16:53,656
BF BINDS ISO METRICALLY AND THIS

2612
02:16:53,656 --> 02:17:03,967
IS PRETTY OBVIOUS.

2613
02:17:12,675 --> 02:17:13,977
WE LOOKED AT THE INTERACTION AND

2614
02:17:13,977 --> 02:17:16,746
THE RESIDUES PARTICIPATING HERE 

2615
02:17:16,746 --> 02:17:23,686
ARE HIGHLIGHTED.

2616
02:17:23,686 --> 02:17:24,754
BASED ON THESE CORRELATIONS THEY

2617
02:17:24,754 --> 02:17:27,590
WERE ABILITY TO DETERMINE THE 

2618
02:17:27,590 --> 02:17:30,593
BINDING MOLD AND YOU CAN SEE THE

2619
02:17:30,593 --> 02:17:32,295
SMALLER MOLECULE ACTING ON THE 

2620
02:17:32,295 --> 02:17:36,833
CHAINS AND THE IP6 AND LED TO 

2621
02:17:36,833 --> 02:17:38,067
DIFFERENT ORIENTATION.

2622
02:17:38,067 --> 02:17:39,469
ONE THING UNIQUE AND UNEXPECTED 

2623
02:17:39,469 --> 02:17:44,007
IN OUR STUDY WAS THAT WE WERE 

2624
02:17:44,007 --> 02:17:49,279
ABLE TO ASSIGN BIASED ON THE 

2625
02:17:49,279 --> 02:17:52,615
SPECTRA FOR THE RAS ARE 

2626
02:17:52,615 --> 02:17:52,882
DIFFERENT.

2627
02:17:52,882 --> 02:17:58,755
THIS COULD SERVE AS A QUICK 

2628
02:17:58,755 --> 02:18:09,265
PROBE OF ENAN TIMER STRUCTURE 

2629
02:18:19,208 --> 02:18:22,745
AND IT'S AN INHIBITOR AND 

2630
02:18:22,745 --> 02:18:25,014
STARTED WITH ERIC AND COLLEAGUES

2631
02:18:25,014 --> 02:18:27,884
LOOKING AT THE SECOND GENERATION

2632
02:18:27,884 --> 02:18:31,120
MATURATION OF INHIBITERS AND 

2633
02:18:31,120 --> 02:18:33,423
ANALOGS OF BVM AND THEY HAVE 

2634
02:18:33,423 --> 02:18:34,757
DIFFERENT GROUPS AND SLIGHTLY 

2635
02:18:34,757 --> 02:18:44,901
LONGER.

2636
02:18:46,402 --> 02:18:49,739
AND THERE WAS THE NUCLEIC ACID 

2637
02:18:49,739 --> 02:18:52,675
DOMAIN AND THERE'S A 

2638
02:18:52,675 --> 02:18:53,876
STABILIZATION OF THE SP 1 TAIL 

2639
02:18:53,876 --> 02:18:57,447
AND THE NC RESIDUES.

2640
02:18:57,447 --> 02:18:59,983
WE'RE WORKING TOWARDS TO THE 

2641
02:18:59,983 --> 02:19:00,683
STRUCTURE AND HAVE CLUES WHAT 

2642
02:19:00,683 --> 02:19:01,551
HAPPENS AS FAR AS THE BINDING 

2643
02:19:01,551 --> 02:19:04,787
AND HOPEFULLY TELL YOU IN THE 

2644
02:19:04,787 --> 02:19:15,098
NEXT YEAR OR TWO.

2645
02:19:16,833 --> 02:19:19,502
AND WE CAN GET ATOMIC RESOLUTION

2646
02:19:19,502 --> 02:19:21,738
STRUCTURES OF PROTEINS 

2647
02:19:21,738 --> 02:19:25,975
ASSEMBLIES AND COMPLEXES WITH 

2648
02:19:25,975 --> 02:19:28,211
SMALL MOLECULES AND WE DON'T 

2649
02:19:28,211 --> 02:19:32,849
NEED TO DO ASYMMETRY AND WE'RE 

2650
02:19:32,849 --> 02:19:34,183
WORKING TOWARDS UNDERSTANDING 

2651
02:19:34,183 --> 02:19:37,153
HOW THE UNIQUE SECOND GENERATION

2652
02:19:37,153 --> 02:19:41,791
IN WHY INHIBITORS TO THE HELIX 

2653
02:19:41,791 --> 02:19:44,694
BUNDLE AND I WANT TO ONCE AGAIN 

2654
02:19:44,694 --> 02:19:46,562
THANK THE PEOPLE WHO HAVE BEEN 

2655
02:19:46,562 --> 02:19:47,130
INVOLVED IN THE WORK.

2656
02:19:47,130 --> 02:19:48,931
I CAN'T STRESS ENOUGH HOW 

2657
02:19:48,931 --> 02:19:50,867
IMPORTANT THE CENTERS ARE TO 

2658
02:19:50,867 --> 02:19:55,905
PROMOTE THESE KINDS OF 

2659
02:19:55,905 --> 02:19:56,973
INTEGRATIVE COLLABORATIVE 

2660
02:19:56,973 --> 02:19:57,640
STUDIES AND THANK YOU VERY MUCH 

2661
02:19:57,640 --> 02:20:07,884
FOR LISTENING.

2662
02:20:22,065 --> 02:20:28,404
>> SO I'LL TALK ABOUT
RANBP2 AND

2663
02:20:28,404 --> 02:20:29,572
IT LOCALIZES TO THE NUCLEAR CORE

2664
02:20:29,572 --> 02:20:32,008
COMPLEX AT THE END TERMINUS.

2665
02:20:32,008 --> 02:20:36,579
AND IT INHIBITS THE INTEGRATION 

2666
02:20:36,579 --> 02:20:38,614
COMPLEX OF HIV AND OTHER PRIMATE

2667
02:20:38,614 --> 02:20:41,984
LENTI VIRUSES BECAUSE OF THIS 

2668
02:20:41,984 --> 02:20:44,854
THE VIRAL CAPSID IS THE 

2669
02:20:44,854 --> 02:20:46,289
DETERMINATE FOR SENSITIVITY AND 

2670
02:20:46,289 --> 02:20:51,394
CAPSID INTERACTION A MAJOR DE 

2671
02:20:51,394 --> 02:20:53,896
TERMINUS AND THIS IS SENSITIVE 

2672
02:20:53,896 --> 02:20:57,800
TO MX2 BUT IF WE HAVE A CAPSID 

2673
02:20:57,800 --> 02:21:01,270
MUTANT THAT DOESN'T BIND OR 

2674
02:21:01,270 --> 02:21:03,506
KNOCKOUT CELLS OR CYCLOSPORINE 

2675
02:21:03,506 --> 02:21:05,708
IT'S SOMETIME ENHANCED IN THESE 

2676
02:21:05,708 --> 02:21:06,008
SITUATIONS.

2677
02:21:06,008 --> 02:21:08,745
BUT THIS IS ONLY TRUE FOR HIV-1 

2678
02:21:08,745 --> 02:21:10,413
AND NOT HIV-2.

2679
02:21:10,413 --> 02:21:14,417
CONTROL CELLS BOTH VIRUS 

2680
02:21:14,417 --> 02:21:17,620
SENSITIVE TO MX2 BUT IT REMAINS 

2681
02:21:17,620 --> 02:21:25,995
SENSITIVE EVEN WHEN NOT BOUND TO

2682
02:21:25,995 --> 02:21:29,932
PSYCYCLOPHILIN AND ONE OF THE 

2683
02:21:29,932 --> 02:21:33,970
QUESTIONS WE'RE INTERESTED IS 

2684
02:21:33,970 --> 02:21:37,974
WHAT ARE INVOLVED IN THE PORE OF

2685
02:21:37,974 --> 02:21:40,810
AND THE ABILITY TO INHIBIT 

2686
02:21:40,810 --> 02:21:45,047
INFECTION.

2687
02:21:45,047 --> 02:21:46,749
STUDIES HAVE DEMONSTRATED 

2688
02:21:46,749 --> 02:21:52,722
INTERACTIONS BETWEEN THE CAPSID 

2689
02:21:52,722 --> 02:21:58,294
BUT THEY MAY NOT HAVE FUNCTIONAL

2690
02:21:58,294 --> 02:22:00,596
CONSEQUENCES AND WHEN WE KNOCKED

2691
02:22:00,596 --> 02:22:02,265
DOWN CERTAIN METHODS THERE WAS 

2692
02:22:02,265 --> 02:22:05,134
AN INDICATION THEY'RE PLAYING A 

2693
02:22:05,134 --> 02:22:09,972
ROLE BUT WILL THERE COULD BE 

2694
02:22:09,972 --> 02:22:10,840
CONFOUNDING RESULTS.

2695
02:22:10,840 --> 02:22:12,375
HOW CAN WE DETERMINE WHAT THEY 

2696
02:22:12,375 --> 02:22:22,451
ARE?

2697
02:22:24,253 --> 02:22:25,788
SRNA COULD MESS UP THE PROBLEM 

2698
02:22:25,788 --> 02:22:28,825
AND IF YOU DO MORE PRECISE 

2699
02:22:28,825 --> 02:22:32,762
ENGINEERING USING A CRISPR 9 AD 

2700
02:22:32,762 --> 02:22:33,996
APPROACH YOU CAN SEPARATE THEM 

2701
02:22:33,996 --> 02:22:36,866
FROM IT THEIR DOMAIN DELETED.

2702
02:22:36,866 --> 02:22:38,835
I WANT TO HIGHLIGHT THE WORK HAS

2703
02:22:38,835 --> 02:22:40,837
DIDN'T DONE BY TWO OUTSTANDING 

2704
02:22:40,837 --> 02:22:45,474
TECHNICIANS IN MY LAB HAILEY AND

2705
02:22:45,474 --> 02:22:51,714
ANANYA AND WE'RE FOCUSSED ON THE

2706
02:22:51,714 --> 02:22:57,353
PEN

2707
02:22:57,353 --> 02:23:00,056
PENTIM

2708
02:23:00,056 --> 02:23:03,793
PENTIMERS THIS BINDS THE VIRAL 

2709
02:23:03,793 --> 02:23:05,928
CAPSID AND WHEN WE KNOCK IT DOWN

2710
02:23:05,928 --> 02:23:06,963
YOU SEE THIS NICE RING AROUND 

2711
02:23:06,963 --> 02:23:08,798
THE NUCLEUS BUT WHEN YOU KNOCK 

2712
02:23:08,798 --> 02:23:10,499
IT DOWN IT'S MISLOCALIZED AND NO

2713
02:23:10,499 --> 02:23:16,472
LONGER INHIBITING INFECTION.

2714
02:23:16,472 --> 02:23:27,016
BECAUSE OF THESE THE EXONS AND 

2715
02:23:30,620 --> 02:23:31,888
WHAT THEY LOOK LIKE AND A 

2716
02:23:31,888 --> 02:23:33,055
WESTERN SHOWING WE GET THE SMALL

2717
02:23:33,055 --> 02:23:33,956
REDUCTION IN THE SIZE OF THE 

2718
02:23:33,956 --> 02:23:41,564
PROTEIN.

2719
02:23:41,564 --> 02:23:43,466
WHEN WE LOOK AT THE CELLS YOU 

2720
02:23:43,466 --> 02:23:48,371
CAN SEE BOTH PROTEINS ARE AT THE

2721
02:23:48,371 --> 02:23:50,172
NUCLEAR ENVELOPE AND IT'S ONE OF

2722
02:23:50,172 --> 02:23:52,541
THE FIRST NUCLEAR CORE TO GO 

2723
02:23:52,541 --> 02:24:03,352
THERE AT THEN OF MITOMITOSIS.

2724
02:24:06,555 --> 02:24:09,492
AND THEY BIND AS THE WILL GD9V 

2725
02:24:09,492 --> 02:24:12,895
AND ALL THREE CAPSIDS ALSO STILL

2726
02:24:12,895 --> 02:24:15,765
BIND WHEN LACKING THE 

2727
02:24:15,765 --> 02:24:19,035
CYCLOPHILIN HOMOLOGY AND SIT 

2728
02:24:19,035 --> 02:24:21,070
INDEPENDENT BETWEEN RANBP2 AND 

2729
02:24:21,070 --> 02:24:25,942
THE CAPSID AND WHEN WE LOOK AT 

2730
02:24:25,942 --> 02:24:29,845
THE INFECTIVITY MX2 WITH AND 

2731
02:24:29,845 --> 02:24:30,613
WITHOUT WE SEE INTERESTING 

2732
02:24:30,613 --> 02:24:30,813
THING.

2733
02:24:30,813 --> 02:24:36,652
A SMALL BUT REPRODUCIBLE IN 

2734
02:24:36,652 --> 02:24:39,221
HIV-1 AND LACK OF REPRODUCTION 

2735
02:24:39,221 --> 02:24:43,759
IN THE CELLS IN CONTRAST TO 

2736
02:24:43,759 --> 02:24:45,661
HIV-2 MORE SENSITIVE THAN HIV-1 

2737
02:24:45,661 --> 02:24:47,596
BUT REMAINS SENSITIVE TO MX2 IN 

2738
02:24:47,596 --> 02:24:49,332
THE CELLS.

2739
02:24:49,332 --> 02:24:51,267
WE CAN TAKE THE CONCLUSION OF 

2740
02:24:51,267 --> 02:24:53,569
THE SENSITIVITY ONE STEP FURTHER

2741
02:24:53,569 --> 02:24:56,405
AND THE DOMAIN IS REQUIRED FOR 

2742
02:24:56,405 --> 02:24:57,673
HIV-1 ONE TO BE SENSITIVE BUT 

2743
02:24:57,673 --> 02:25:03,546
NOT HIV-2.

2744
02:25:03,546 --> 02:25:14,090
WE NEXT ASKED IF THE DELETION OF

2745
02:25:18,995 --> 02:25:21,497
THE DOMAIN HAD SIMILAR EFFECTS 

2746
02:25:21,497 --> 02:25:23,265
WE SEE AN INCREASE IN 

2747
02:25:23,265 --> 02:25:27,436
INTEGRATION IN LAMIN ASSOCIATED 

2748
02:25:27,436 --> 02:25:28,904
DOMAINS AND INCREASE IN GENE 

2749
02:25:28,904 --> 02:25:32,174
DENSE REGIONS.

2750
02:25:32,174 --> 02:25:32,842
AS FAR AS INTEGRATION SITE 

2751
02:25:32,842 --> 02:25:33,576
SELECTION THEY APPEAR TO PLAY A 

2752
02:25:33,576 --> 02:25:43,819
SIMILAR ROLE.

2753
02:25:56,866 --> 02:26:00,536
AND WE TOOK YOU'RE CONTROL AND 

2754
02:26:00,536 --> 02:26:02,438
TRANSDUCED THEM WITH OUR MX2 AND

2755
02:26:02,438 --> 02:26:06,709
TRANS FECTED THEM WITH AND ADDED

2756
02:26:06,709 --> 02:26:08,978
THE EXPRESSION AND INFECTED WITH

2757
02:26:08,978 --> 02:26:13,315
THE PURO VIRUS AND QUANTIFIED 

2758
02:26:13,315 --> 02:26:19,188
INFECTIVITY AND ON THE FAR RIGHT

2759
02:26:19,188 --> 02:26:21,991
ARE THE CONTROL CELLS WITH AND 

2760
02:26:21,991 --> 02:26:23,926
WITHOUT MX2 AND SOME REQUIRE 

2761
02:26:23,926 --> 02:26:30,766
VIRAL INFECTION AND ACTIVITY.

2762
02:26:30,766 --> 02:26:32,735
AND THESE ARE IN CONTROL CELLS 

2763
02:26:32,735 --> 02:26:34,537
AND SEE A DIFFERENT PICTURE 

2764
02:26:34,537 --> 02:26:37,973
LOOKING AT THE DEL SIP CELLS.

2765
02:26:37,973 --> 02:26:39,408
THESE MEMBERS ARE OF THE 

2766
02:26:39,408 --> 02:26:39,642
COMPLEX.

2767
02:26:39,642 --> 02:26:42,211
THE VIRUS BECOMES MORE SENSITIVE

2768
02:26:42,211 --> 02:26:44,580
TO DEPLETION OF THIS WHEN IT'S 

2769
02:26:44,580 --> 02:26:48,150
GONE AND WE ALSO SEE INTERESTING

2770
02:26:48,150 --> 02:26:55,691
THINGS WITH THE SIGHT J JO  -- 

2771
02:26:55,691 --> 02:26:57,993
CYTOPLASMIC MOUSE AND THIS IS 

2772
02:26:57,993 --> 02:27:00,696
WHAT WE SEE WITH GV9V INFECTION 

2773
02:27:00,696 --> 02:27:03,933
IN OUR CONTROL CELLS.

2774
02:27:03,933 --> 02:27:04,867
WE SEE REDUCTION UPON DEPLETION 

2775
02:27:04,867 --> 02:27:07,470
OF THE KNOBS RESCUED BY MX2 

2776
02:27:07,470 --> 02:27:12,007
EXPRESSION AND SEE A SIMILAR 

2777
02:27:12,007 --> 02:27:14,276
EFFECT OF THE 155 DEPLETION AND 

2778
02:27:14,276 --> 02:27:17,980
THINK THE CAPSID INTERACTIONS 

2779
02:27:17,980 --> 02:27:20,549
ARE AFFECTING THE INFECTION AND 

2780
02:27:20,549 --> 02:27:21,450
SENSITIVITY BUT THIS IS A LITTLE

2781
02:27:21,450 --> 02:27:23,319
BIT AT THE END SO WHAT ARE THE 

2782
02:27:23,319 --> 02:27:23,919
OTHER DOMAINS DOING FOR VIRAL 

2783
02:27:23,919 --> 02:27:33,963
INFECTION?

2784
02:27:33,963 --> 02:27:36,165
AND THIS IS THE DOMAIN 

2785
02:27:36,165 --> 02:27:37,366
ARCHITECTURE AND WE SEE AN 

2786
02:27:37,366 --> 02:27:39,001
PICTURE LOOKING AT HIV INFECTION

2787
02:27:39,001 --> 02:27:40,002
IN CELLS.

2788
02:27:40,002 --> 02:27:50,479
WE SAW A SMALLER EFFECT ON 

2789
02:27:55,084 --> 02:27:56,652
INFECTIVITY AND THIS IS THE DE 

2790
02:27:56,652 --> 02:27:58,354
MAIN ARCHITECTURE AND NOW WE SEE

2791
02:27:58,354 --> 02:28:00,456
WE HAVE NO EFFECT ON INFECTIVITY

2792
02:28:00,456 --> 02:28:01,957
OR SENSITIVITY OF THE VIRUS IN 

2793
02:28:01,957 --> 02:28:02,625
THE CELLS.

2794
02:28:02,625 --> 02:28:04,059
SO THIS IS TELLING US THERE ARE 

2795
02:28:04,059 --> 02:28:08,197
COMPLEX INTERACTS BETWEEN THE C 

2796
02:28:08,197 --> 02:28:09,565
TERMINAL DOMAIN THAT AFFECT 

2797
02:28:09,565 --> 02:28:10,099
HIV-1 INFECTION AND MX2 

2798
02:28:10,099 --> 02:28:20,309
SENSITIVITY.

2799
02:28:31,086 --> 02:28:34,056
WE SEE SMALL EFFECT BUT MX2 

2800
02:28:34,056 --> 02:28:37,793
ACTIVITY IS ABROGATED AND WHILE 

2801
02:28:37,793 --> 02:28:40,930
IT'S SENSITIVE TO THE IT 

2802
02:28:40,930 --> 02:28:45,301
CYCLOPHILIN HOMOLOGY IT'S A NO 

2803
02:28:45,301 --> 02:28:47,369
THE WHEN YOU GET RID OF THIS AND

2804
02:28:47,369 --> 02:28:52,141
SENSITIVE TO MX2 AND LOOK AT 

2805
02:28:52,141 --> 02:28:53,809
LOCALIZATION OF THE CELLS AROUND

2806
02:28:53,809 --> 02:28:57,012
RANBP2 STILL GOES TO THE NUCLEAR

2807
02:28:57,012 --> 02:28:58,914
ENVELOPE BUT MX2 IS LOCALIZED 

2808
02:28:58,914 --> 02:29:02,151
AND THINK THE RAMIFICATIONS ARE 

2809
02:29:02,151 --> 02:29:03,485
AFFECTING THROUGH DIRECT 

2810
02:29:03,485 --> 02:29:05,955
INTERACTION THE VIRAL CAPSID AND

2811
02:29:05,955 --> 02:29:09,058
INDIRECTLY THROUGH THE MX2 

2812
02:29:09,058 --> 02:29:09,959
LOCALIZATION.

2813
02:29:09,959 --> 02:29:13,028
IN CONTROL CELLS HIV-1 AND HIV-2

2814
02:29:13,028 --> 02:29:19,235
ARE SENSITIVE TO MX2 AND HIV-1 

2815
02:29:19,235 --> 02:29:23,672
IS RESISTANT AND HIV-2 IS 

2816
02:29:23,672 --> 02:29:25,574
NEGATIVE AND WHEN WE REMOVE BOTH

2817
02:29:25,574 --> 02:29:29,245
BOTH ARE SENSITIVE.

2818
02:29:29,245 --> 02:29:31,247
AGAIN THE INTERACTIONS ARE 

2819
02:29:31,247 --> 02:29:41,790
COMPLEX BETWEEN THE CAPSID AND 

2820
02:29:43,859 --> 02:29:44,593
MX2 SENSITIVITY AND THINK THE 

2821
02:29:44,593 --> 02:29:47,496
FOURTH BINDING DOMAIN PLAYS AN 

2822
02:29:47,496 --> 02:29:50,966
INDIRECT ROLE AND INSPENCEABLE 

2823
02:29:50,966 --> 02:29:52,234
FOR HIV INFECTION.

2824
02:29:52,234 --> 02:29:54,470
AND WE INDICATE IT'S INTERACTING

2825
02:29:54,470 --> 02:29:57,339
WITH THE CAPSID AND FG 

2826
02:29:57,339 --> 02:29:59,875
INDEPENDENT MANNER BUT THE 

2827
02:29:59,875 --> 02:30:01,710
EXPRESSION OF THE TRUNCATED 

2828
02:30:01,710 --> 02:30:04,346
RANBP2 IS REQUIRED AND MX2 IS 

2829
02:30:04,346 --> 02:30:05,948
REQUIRED FOR NUCLEAR ENVELOPE 

2830
02:30:05,948 --> 02:30:09,251
LOCALIZING AND ANTIVIRAL 

2831
02:30:09,251 --> 02:30:09,518
ACTIVITY.

2832
02:30:09,518 --> 02:30:11,687
AND WITH THAT I WANT TO THANK 

2833
02:30:11,687 --> 02:30:14,990
EVERYBODY IN MY LAB AND ANANYA 

2834
02:30:14,990 --> 02:30:19,328
IS GOING TO MADISON FOR GRAD 

2835
02:30:19,328 --> 02:30:21,297
SCHOOL THIS FALL AND WILL BE 

2836
02:30:21,297 --> 02:30:23,565
MISSED AND THANK YOU FOR THE 

2837
02:30:23,565 --> 02:30:24,400
FUNDING AND IT'S BEEN GREAT TO 

2838
02:30:24,400 --> 02:30:34,777
BE PART OF THIS GROUP.

2839
02:30:35,044 --> 02:30:36,478
>> OKAY.

2840
02:30:36,478 --> 02:30:44,787
SO I'M NOW STANDING BETWEEN YOU 

2841
02:30:44,787 --> 02:30:45,988
AND MEALS.

2842
02:30:45,988 --> 02:30:47,790
I'LL TRY TO SPEAK AS FAST AS 

2843
02:30:47,790 --> 02:30:48,090
MELISSA.

2844
02:30:48,090 --> 02:30:50,759
SO WHAT I'M TRYING TO STUDY HERE

2845
02:30:50,759 --> 02:30:52,928
IS APPLICATION OF A SINGLE 

2846
02:30:52,928 --> 02:30:59,068
PARTICLE CRYO EM AND NOT CRYO-ED

2847
02:30:59,068 --> 02:31:01,403
FOR IN SITU STUDIES IN A NATIVE 

2848
02:31:01,403 --> 02:31:01,904
SELL ENVIRONMENT.

2849
02:31:01,904 --> 02:31:05,274
FOR THE FIRST PART OF MY TALK 

2850
02:31:05,274 --> 02:31:11,313
I'LL TALK BRIEFLY ON VIRUS WORK 

2851
02:31:11,313 --> 02:31:15,818
BUT THIS IS THE TREND IN THE 

2852
02:31:15,818 --> 02:31:17,619
FIELD AS YOU HEARD ABOUT 

2853
02:31:17,619 --> 02:31:21,090
YESTERDAY AND TALK ABOUT 

2854
02:31:21,090 --> 02:31:22,891
STRUCTURAL BIOLOGY AND MY LAB IS

2855
02:31:22,891 --> 02:31:26,662
ALSO LOOKING AT THE SINGLE 

2856
02:31:26,662 --> 02:31:35,571
PARTICLE CRY SCRO -- CRYO EM 

2857
02:31:35,571 --> 02:31:37,272
FROM MAMMALIAL CELLS AND GET A 

2858
02:31:37,272 --> 02:31:40,909
RANGE AND THIS WORK IS 

2859
02:31:40,909 --> 02:31:43,846
SPEARHEADED AND CO-LED BY MY LAB

2860
02:31:43,846 --> 02:31:46,448
WITH HELP WITH RACHEL AND FOR 

2861
02:31:46,448 --> 02:31:48,183
THE INTEREST OF TIME I WILL ONLY

2862
02:31:48,183 --> 02:31:52,888
PRESENT ONE STRUCTURE TO YOU 

2863
02:31:52,888 --> 02:31:56,892
TODAY OF THIS CAREFUL VIRUS.

2864
02:31:56,892 --> 02:31:58,894
SO WHY THE TREATMENT?

2865
02:31:58,894 --> 02:32:01,063
WHAT IS PFO.

2866
02:32:01,063 --> 02:32:04,233
IT'S BACTERIAL TOXINS ON THE 

2867
02:32:04,233 --> 02:32:05,667
LIPID MEMBRANE.

2868
02:32:05,667 --> 02:32:11,740
THIS IS A FIRST APPLIED TO THE 

2869
02:32:11,740 --> 02:32:18,280
MATURE VIRAL VLPs AND LABS USE 

2870
02:32:18,280 --> 02:32:21,049
THIS FOR CRYO EM WORK AND THIS 

2871
02:32:21,049 --> 02:32:25,721
IS MATURE CAPSIDS AND CAN WE 

2872
02:32:25,721 --> 02:32:26,789
APPLY THIS BECAUSE IT'S CLOSE TO

2873
02:32:26,789 --> 02:32:27,289
THE MEMBRANE.

2874
02:32:27,289 --> 02:32:32,060
DOES IT WORK FOR TREATMENT?

2875
02:32:32,060 --> 02:32:33,395
AND YOU CAN COMPARE FROM A 

2876
02:32:33,395 --> 02:32:36,231
COMPARISON OF THE BEFORE AND 

2877
02:32:36,231 --> 02:32:37,433
AFTER TREATMENT YOU CAN CLEARLY 

2878
02:32:37,433 --> 02:32:39,835
SEE THE PORES FORM AS INDICATED 

2879
02:32:39,835 --> 02:32:41,370
BY ARROWS AND WE'RE VERY HAPPY 

2880
02:32:41,370 --> 02:32:46,442
TO SEE IT WORKS ON THE IMMATURE 

2881
02:32:46,442 --> 02:32:49,978
VLP AND IT APPEARS TO BE INTACT.

2882
02:32:49,978 --> 02:32:53,549
IT'S NOW DISRUPTING THE LATTICE.

2883
02:32:53,549 --> 02:32:56,351
IT POKE HOLE TO HELP INFLUX OF 

2884
02:32:56,351 --> 02:33:01,223
DRUG OR INHIBITOR PRECURSORS SO 

2885
02:33:01,223 --> 02:33:03,459
WHETHER THEY'RE OPTIMIZED WILL 

2886
02:33:03,459 --> 02:33:04,726
HELP WITH PERMEABILITY.

2887
02:33:04,726 --> 02:33:09,264
WE USE THIS TO STUDY A FEW 

2888
02:33:09,264 --> 02:33:10,098
DIFFERENT DRUG COMPOUNDS WHEREAS

2889
02:33:10,098 --> 02:33:13,368
I SAID I'LL ONLY TALK TO YOU 

2890
02:33:13,368 --> 02:33:15,737
ABOUT A DOUBLE TREATED VLP.

2891
02:33:15,737 --> 02:33:20,676
SO IT WORKED WELL AND GOT HIGH 

2892
02:33:20,676 --> 02:33:26,014
RESOLUTION OF 2.1 IN RESOLUTION 

2893
02:33:26,014 --> 02:33:29,184
AND LOOKS AT THE BVM AND YOU SEE

2894
02:33:29,184 --> 02:33:32,387
NICE IP6 ON TOP OF THE BUNDLE OF

2895
02:33:32,387 --> 02:33:36,692
CSP1 AND FROM THE SIDE VIEW YOU 

2896
02:33:36,692 --> 02:33:40,896
SEE THE IG6 AND BVM IS WELL 

2897
02:33:40,896 --> 02:33:43,832
DEFINED DENSITY.

2898
02:33:43,832 --> 02:33:45,501
WE KNOW PRECISELY THE LOCATION 

2899
02:33:45,501 --> 02:33:46,134
AND CAN FIT A NICE MODEL INTO 

2900
02:33:46,134 --> 02:33:51,173
IT.

2901
02:33:51,173 --> 02:33:57,346
HOW ABOUT LENACAPAVIR AND IF I 

2902
02:33:57,346 --> 02:34:04,887
TAKE THIS AND TURN IT 90 DEGREE 

2903
02:34:04,887 --> 02:34:09,124
VIEW AND IV6 THE LENACAPAVIR 

2904
02:34:09,124 --> 02:34:11,960
BINDS TO NTD ONLY IN THE 

2905
02:34:11,960 --> 02:34:14,129
IMMATURE VIRION AND WE HAVE GOOD

2906
02:34:14,129 --> 02:34:16,532
DENSITY TO DEFINE THE FUNCTIONAL

2907
02:34:16,532 --> 02:34:18,233
GROUPS AND IT DOES ACTUALLY HAVE

2908
02:34:18,233 --> 02:34:21,970
A DIFFERENT POST COMPARED TO THE

2909
02:34:21,970 --> 02:34:23,005
MATURE LENACAPAVIR BINDING TO 

2910
02:34:23,005 --> 02:34:24,406
THE MATURE CAPSID.

2911
02:34:24,406 --> 02:34:28,877
AND THE BINDING ACTUALLY CAUSES 

2912
02:34:28,877 --> 02:34:30,979
A CONFIRMATIONAL CHANGE PART OF 

2913
02:34:30,979 --> 02:34:31,547
THE LAG.

2914
02:34:31,547 --> 02:34:34,283
IT'S A RELATIVE ORIENTATION OF 

2915
02:34:34,283 --> 02:34:38,120
THE NTCA RELATIVE TO THE CTD.

2916
02:34:38,120 --> 02:34:41,256
AS A CONSEQUENCE WE SEE A 

2917
02:34:41,256 --> 02:34:44,259
MORPHING MOTION OF THE LATTICE 

2918
02:34:44,259 --> 02:34:49,031
TYPE AND THE IMMATURE LATTICE IS

2919
02:34:49,031 --> 02:34:50,098
MALLEABLE AND CAN CHANGE.

2920
02:34:50,098 --> 02:34:54,202
AS A QUICK SUMMARY OF THIS PART,

2921
02:34:54,202 --> 02:34:56,038
PFO TREATMENT WORKS FOR INHI

2922
02:34:56,038 --> 02:34:57,873
INHIBITOR STUDIES AND ALLOWED US

2923
02:34:57,873 --> 02:35:00,442
TO OBTAIN A HIGH RESOLUTION 

2924
02:35:00,442 --> 02:35:02,544
STRUCTURE OF LENACAPAVIR AND 

2925
02:35:02,544 --> 02:35:03,645
IDENTIFY A NEW LOCATION AND POST

2926
02:35:03,645 --> 02:35:07,316
OF BINDING AND AT THE SAME TIME 

2927
02:35:07,316 --> 02:35:10,152
WITH GOOD DEFINITION OF BBM 

2928
02:35:10,152 --> 02:35:11,920
INDICATING THEY CAN WORK 

2929
02:35:11,920 --> 02:35:12,187
TOGETHER.

2930
02:35:12,187 --> 02:35:17,993
AND ALSO WE IDENTIFIED AN 

2931
02:35:17,993 --> 02:35:21,763
IMMATURE CAPSID MAY HAVE 

2932
02:35:21,763 --> 02:35:22,431
SIGNIFICANT INDICATIONS IN THE 

2933
02:35:22,431 --> 02:35:27,135
PATHWAY OF THE CAPSID.

2934
02:35:27,135 --> 02:35:30,005
SO THE STRUCTURAL BIOLOGY YOU 

2935
02:35:30,005 --> 02:35:31,206
HAVE SINGLE BIOLOGY CRYO EM WORK

2936
02:35:31,206 --> 02:35:33,976
AND WHAT WORKS WELL FOR VIRUSES 

2937
02:35:33,976 --> 02:35:35,777
HAS GREAT OUTCOMES AND I WANT TO

2938
02:35:35,777 --> 02:35:39,982
SPEND MORE TIME WITH YOU ON THE 

2939
02:35:39,982 --> 02:35:41,783
HIGH RESOLUTION RESOLUTION IN 

2940
02:35:41,783 --> 02:35:42,784
CELLS BASICALLY FROM TEST TUBE 

2941
02:35:42,784 --> 02:35:43,919
TO CELLS.

2942
02:35:43,919 --> 02:35:46,188
THIS IS REALLY A DREAM FOR MANY 

2943
02:35:46,188 --> 02:35:52,694
OF US FOR DECADES AND ALSO 

2944
02:35:52,694 --> 02:35:56,598
ACCELERATED BY THE MAJOR 

2945
02:35:56,598 --> 02:35:58,934
DEVELOPMENT IN THE FIELD 

2946
02:35:58,934 --> 02:36:01,970
BASICALLY THE THINNING THE CELL 

2947
02:36:01,970 --> 02:36:05,974
SAMPLE BY CRYO ET AS ALBERTO 

2948
02:36:05,974 --> 02:36:11,146
NICELY ILLUSTRATED IN THE 

2949
02:36:11,146 --> 02:36:16,685
MORNING.

2950
02:36:16,685 --> 02:36:19,421
AS NICE AS IT IS DUE TO 

2951
02:36:19,421 --> 02:36:21,456
INTRINSIC LIMITATIONS AND THE 

2952
02:36:21,456 --> 02:36:22,457
HIGH LEVEL OF EXPERTISE NEEDED 

2953
02:36:22,457 --> 02:36:32,701
TO DO IT WELL.

2954
02:36:33,135 --> 02:36:35,337
AND YOU CAN TAKE A SINGLE 

2955
02:36:35,337 --> 02:36:38,206
PARTICLE TYPE OF WORK WITHOUT 

2956
02:36:38,206 --> 02:36:40,976
ANY TILT CAN WE DO HIGH 

2957
02:36:40,976 --> 02:36:49,351
RESOLUTION STRUCTURES.

2958
02:36:49,351 --> 02:36:54,990
THIS IS SPEARHEADED BY OUR 

2959
02:36:54,990 --> 02:36:56,024
POSTDOC IN THE LAB.

2960
02:36:56,024 --> 02:36:57,993
I'LL WALK YOU THROUGH THE 

2961
02:36:57,993 --> 02:36:58,260
PIPELINE.

2962
02:36:58,260 --> 02:37:01,963
ON THE LEFT ARE FROZEN CELLS 

2963
02:37:01,963 --> 02:37:09,971
POSITIONED TO BE MARK AND FOR 

2964
02:37:09,971 --> 02:37:13,842
THE TALK THE WORK I'LL PRESENT 

2965
02:37:13,842 --> 02:37:15,677
TODAY I HAVE TWO DATA SETS ON 

2966
02:37:15,677 --> 02:37:17,946
NATIVE UNTREATED CELL AND THE 

2967
02:37:17,946 --> 02:37:22,684
INHIBITOR TREATED CELL.

2968
02:37:22,684 --> 02:37:29,658
THEN WE ACTUAL LY ACTUALLY HAD  

2969
02:37:29,658 --> 02:37:31,226
BOTTLENECK ON THE WORK SO WE 

2970
02:37:31,226 --> 02:37:33,395
NEED BETTER EQUIPMENT AND THE 

2971
02:37:33,395 --> 02:37:36,698
TRAINING AND EXPERIMENTING TO 

2972
02:37:36,698 --> 02:37:39,067
GET THERE TAKES ABOUT 100 DAYS.

2973
02:37:39,067 --> 02:37:40,268
THIS WOULD HAVE BEEN IMPOSSIBLE 

2974
02:37:40,268 --> 02:37:42,971
WITHOUT THE GREAT SUPPORT OF OUR

2975
02:37:42,971 --> 02:37:46,942
COLLEAGUES AT YALE ESPECIALLY 

2976
02:37:46,942 --> 02:37:52,681
JUN LU WHO REALLY TOLERATED US 

2977
02:37:52,681 --> 02:37:58,587
WHO TRAINED US TO DO IT WELL.

2978
02:37:58,587 --> 02:38:00,589
IF WE GET OVER THIS HURDLE THE 

2979
02:38:00,589 --> 02:38:05,293
REST IS SMOOTH AND ENJOYABLE.

2980
02:38:05,293 --> 02:38:07,796
WE TOGETHER WITH OUR COLLEAGUES 

2981
02:38:07,796 --> 02:38:09,798
WE DEVELOPED A QUITE EFFICIENT 

2982
02:38:09,798 --> 02:38:11,399
SINGLE PARTICLE PIPELINE LIKE 

2983
02:38:11,399 --> 02:38:13,068
YOU COLLECT PURIFIED SAMPLE IN 

2984
02:38:13,068 --> 02:38:13,969
THE DATA SET.

2985
02:38:13,969 --> 02:38:17,439
YOU CAN DEFINE A FOCUS REGION TO

2986
02:38:17,439 --> 02:38:19,341
DO DATA COLLECTION AND MOVE TO 

2987
02:38:19,341 --> 02:38:21,510
DIFFERENT LOCATIONS TO DO THE 

2988
02:38:21,510 --> 02:38:21,977
SAME.

2989
02:38:21,977 --> 02:38:25,547
SO WITH THIS SCHEME WE CAN GET 

2990
02:38:25,547 --> 02:38:27,282
ABOUT 4,000 TO 5,000 MICROGRAPHS

2991
02:38:27,282 --> 02:38:29,584
PER DAY WHICH IS NOT THE SAME AS

2992
02:38:29,584 --> 02:38:33,955
SINGLE PARTICLES VERY CLOSE TO 

2993
02:38:33,955 --> 02:38:36,625
THE THROUGHPUT PURIFIED SAMPLE.

2994
02:38:36,625 --> 02:38:39,394
THESE ARE THE TYPICAL 

2995
02:38:39,394 --> 02:38:40,362
MICROGRAPHS WE LOOK AT THESE 

2996
02:38:40,362 --> 02:38:40,862
DAYS.

2997
02:38:40,862 --> 02:38:42,931
THE PROOF OF PRINCIPLE STUDIES 

2998
02:38:42,931 --> 02:38:53,408
AND FOCUS ON THE BLACK DOTS.

2999
02:39:05,320 --> 02:39:07,455
AND IT TURNED OUT PAR BETTER 

3000
02:39:07,455 --> 02:39:08,723
THAN WE EXPECT.

3001
02:39:08,723 --> 02:39:11,259
THIS IS A WILL E-MAIL TO THE 

3002
02:39:11,259 --> 02:39:14,896
TEAM THE SECOND DAY AFTER THE 

3003
02:39:14,896 --> 02:39:20,836
INTERACTION AND 3.6 RESOLUTION 

3004
02:39:20,836 --> 02:39:23,071
ENTIRELY USING THE PIPELINE 

3005
02:39:23,071 --> 02:39:25,440
USING CRYO SPARK NO SPECIAL 

3006
02:39:25,440 --> 02:39:28,977
TREATMENT AND ASSEMBLED A LARGE 

3007
02:39:28,977 --> 02:39:34,516
TEAM AND TOOK ADVANTAGE OF THE 

3008
02:39:34,516 --> 02:39:36,484
ALGORITHM DEVELOPED THAT WORKED 

3009
02:39:36,484 --> 02:39:37,252
BETTER.

3010
02:39:37,252 --> 02:39:39,054
A MONTH LATER WE HAD 2.5 

3011
02:39:39,054 --> 02:39:40,989
ASSOCIATION AND THESE ARE THE 

3012
02:39:40,989 --> 02:39:44,626
SEN

3013
02:39:44,626 --> 02:39:51,933
SENSITY WE SAW AND -- 

3014
02:39:51,933 --> 02:39:52,968
SENSITIVITY AND THIS IS THE 

3015
02:39:52,968 --> 02:39:54,436
DENSITY WE HAVE NOW.

3016
02:39:54,436 --> 02:39:57,973
IT'S NICE TO SEE THE AROMATICS 

3017
02:39:57,973 --> 02:40:00,475
AND SMALL SIDE CHAINS AND GOOD 

3018
02:40:00,475 --> 02:40:05,914
DEFINITION WITH THE RNA POST 

3019
02:40:05,914 --> 02:40:16,458
TRANSLATIONAL GROUP ON THE BASE 

3020
02:40:18,827 --> 02:40:26,935
AND WE USE THE INHIBITOR AND WE 

3021
02:40:26,935 --> 02:40:29,604
SEE GOOD DENSITY IN THE CELL.

3022
02:40:29,604 --> 02:40:31,439
HIGH RESOLUTION IS GOOD.

3023
02:40:31,439 --> 02:40:34,776
WE ALSO SEE NICE INTERACTION 

3024
02:40:34,776 --> 02:40:38,980
BETWEEN TRNA AND CODEINE 

3025
02:40:38,980 --> 02:40:40,448
INTERACTION AND IF I DON'T SPEAK

3026
02:40:40,448 --> 02:40:42,317
THIS FAST IT GIVES TIME TO THINK

3027
02:40:42,317 --> 02:40:45,954
AND YOU MAY THINK WAIT A MINUTE 

3028
02:40:45,954 --> 02:40:50,091
AND THIS IS A HIGHLY COMPLEX AND

3029
02:40:50,091 --> 02:40:51,927
DYNAMIC PROCESS.

3030
02:40:51,927 --> 02:40:55,897
CLOSE TO 60TRNAs BRING ACIDS 

3031
02:40:55,897 --> 02:40:57,265
PACKING THROUGH MANY SITES WITH 

3032
02:40:57,265 --> 02:41:01,970
CONFIRMATIONAL CHANGES.

3033
02:41:01,970 --> 02:41:02,570
BASICALLY WE HAVE EVERYTHING 

3034
02:41:02,570 --> 02:41:05,373
EVERYWHERE ALL AT ONCE.

3035
02:41:05,373 --> 02:41:06,274
THIS A CHALLENGE BUT ALSO 

3036
02:41:06,274 --> 02:41:06,608
BLESSING.

3037
02:41:06,608 --> 02:41:08,610
IF WE CAN FIGURE OUT THE 

3038
02:41:08,610 --> 02:41:12,280
DIFFERENT STATES WE LEARN MORE.

3039
02:41:12,280 --> 02:41:16,985
THAT'S WE -- THAT'S WHAT WE DID 

3040
02:41:16,985 --> 02:41:19,154
AND WE USED THE SINGLE PARTICLES

3041
02:41:19,154 --> 02:41:23,024
AND SOME CALL IT PURIFICATION IN

3042
02:41:23,024 --> 02:41:25,961
SILICO AND HERE'S A CLASS OF 

3043
02:41:25,961 --> 02:41:31,666
HIBERNATING RIBOSOME AND HERE'S 

3044
02:41:31,666 --> 02:41:34,869
ANOTHER PTRNA AND BOUND TO THE 

3045
02:41:34,869 --> 02:41:37,973
SIDE AND MORE AND MORE AND MORE 

3046
02:41:37,973 --> 02:41:42,677
AND MORE.

3047
02:41:42,677 --> 02:41:45,680
AND THIS IS A Z tRNA IN THE 

3048
02:41:45,680 --> 02:41:45,880
CELL.

3049
02:41:45,880 --> 02:41:49,951
AT THE END WE HAVE AN EXTENSIVE 

3050
02:41:49,951 --> 02:41:51,920
TRANSLATION CYCLE WITH 21 

3051
02:41:51,920 --> 02:41:56,091
DISTINCT STATES AND ALL BUT ONE 

3052
02:41:56,091 --> 02:41:57,792
IS DETERMINATED IN THE 

3053
02:41:57,792 --> 02:42:00,962
RESOLUTION AND THIS IS 3.3 NOT 

3054
02:42:00,962 --> 02:42:01,830
TOO SHABBY.

3055
02:42:01,830 --> 02:42:04,199
AND ALSO BECAUSE WE HAVE OVER A 

3056
02:42:04,199 --> 02:42:06,735
MILLION HIGH QUALITY PARTICLES 

3057
02:42:06,735 --> 02:42:10,839
WE CAN DO STATISTICS TO FIGURE 

3058
02:42:10,839 --> 02:42:13,975
OUT THE STATE DISTRIBUTION AND 

3059
02:42:13,975 --> 02:42:17,812
THE TRANSLATION LANDSCAPE.

3060
02:42:17,812 --> 02:42:23,218
YOU HAVE TIME TO TILL ABOUT ONE 

3061
02:42:23,218 --> 02:42:29,057
STORY AND CRBP1 AND THIS CENTRAL

3062
02:42:29,057 --> 02:42:31,659
REGION OF THIS LONG NOODLY 

3063
02:42:31,659 --> 02:42:34,562
MOLECULE HAS BEEN OBSERVED 

3064
02:42:34,562 --> 02:42:35,730
OCCUPYING THE mRNA CHANNEL AND 

3065
02:42:35,730 --> 02:42:42,570
PUT IT INTO A NON-TRANSLATING OR

3066
02:42:42,570 --> 02:42:49,944
HIBERNATING STATE AND WE SEE THE

3067
02:42:49,944 --> 02:43:00,488
END TERMINUS AND THIS INDICATES 

3068
02:43:02,957 --> 02:43:13,068
IT'S A POTENT SO TRANSLATE TO 

3069
02:43:13,068 --> 02:43:16,171
HIBERNATION AND MORE STRIKINGLY 

3070
02:43:16,171 --> 02:43:19,374
WE SEE THIS ALWAYS ON THE UNIT 

3071
02:43:19,374 --> 02:43:23,511
WHICH IS A TANTALIZING MODEL 

3072
02:43:23,511 --> 02:43:28,216
THAT MAYBE PERHAPS IN THE CELL 

3073
02:43:28,216 --> 02:43:30,552
EVERYTHING IS COMING AS TETHERED

3074
02:43:30,552 --> 02:43:30,752
PAIRS.

3075
02:43:30,752 --> 02:43:32,253
THIS IS HIGHLY SPECULATIVE.

3076
02:43:32,253 --> 02:43:32,854
WE DON'T HAVE DATA TO SUPPORT 

3077
02:43:32,854 --> 02:43:37,625
THIS.

3078
02:43:37,625 --> 02:43:40,295
TO MORE SLIDES TO SHOW.

3079
02:43:40,295 --> 02:43:45,233
WE HAVE AN INHIBITOR TREATED 

3080
02:43:45,233 --> 02:43:47,202
CELL AND SEEING INTERACTION WE 

3081
02:43:47,202 --> 02:43:49,771
SEE DO TRANSLATION LANDSCAPE 

3082
02:43:49,771 --> 02:43:52,006
ANALYSIS TO SEE HOW THEY CHANGE 

3083
02:43:52,006 --> 02:43:53,007
THE TRANSLATION STATES 

3084
02:43:53,007 --> 02:44:00,882
DISTRIBUTION.

3085
02:44:00,882 --> 02:44:05,954
WE SEE OTHER COFACTORS SUCH AS 

3086
02:44:05,954 --> 02:44:14,996
POLY AMINES AND THIS IS ENGAGED 

3087
02:44:14,996 --> 02:44:19,934
IN HEXAMIDE IS LIKELY FROM 

3088
02:44:19,934 --> 02:44:21,936
PURIFIED SAMPLE STUDY AND IT'S 

3089
02:44:21,936 --> 02:44:22,770
NOT INCLUDED.

3090
02:44:22,770 --> 02:44:24,772
OF COURSE THEY DON'T SEE IT.

3091
02:44:24,772 --> 02:44:27,208
THIS EMPHASIZES TO DO THE WORK 

3092
02:44:27,208 --> 02:44:27,876
IN THE CORRECT ENVIRONMENT AND 

3093
02:44:27,876 --> 02:44:34,716
CELL IS THE BEST YOU CAN GET.

3094
02:44:34,716 --> 02:44:37,285
IN SUMMARY IT HOLDS GREAT 

3095
02:44:37,285 --> 02:44:38,586
POTENTIAL FOR STUDIES BUT THIS 

3096
02:44:38,586 --> 02:44:40,622
IS JUST THE BEGINNING.

3097
02:44:40,622 --> 02:44:41,956
SO MUCH NEEDS TO BE OPTIMIZED 

3098
02:44:41,956 --> 02:44:51,199
AND SO MUCH CAN BE DONE.

3099
02:44:51,199 --> 02:44:53,001
WE CAN DO RIBOSOME RELATED 

3100
02:44:53,001 --> 02:44:56,204
BIOLOGY IN CELLS AND SOME 

3101
02:44:56,204 --> 02:44:57,705
PHARMACOLOGY BUT COMING BACK TO 

3102
02:44:57,705 --> 02:44:59,641
VIROLOGY IS WHERE WE DEVELOPED 

3103
02:44:59,641 --> 02:45:00,642
THE SYSTEM.

3104
02:45:00,642 --> 02:45:08,283
WE CAN STUDY THE VIRAL  

3105
02:45:08,283 --> 02:45:09,884
ALTERATION A COMMON PATHWAY WITH

3106
02:45:09,884 --> 02:45:13,154
THE RIBOSOMES SHIFTING THE 

3107
02:45:13,154 --> 02:45:15,990
EFFECT ON TRANSLATION EFFICIENCY

3108
02:45:15,990 --> 02:45:17,559
AND ALTERNATIVE INITIATION 

3109
02:45:17,559 --> 02:45:17,792
VIRUSES.

3110
02:45:17,792 --> 02:45:22,964
AND WE CAN ALSO STUDY HIV 

3111
02:45:22,964 --> 02:45:24,699
MATURATION TETHERED TO CELL 

3112
02:45:24,699 --> 02:45:24,933
SURFACE.

3113
02:45:24,933 --> 02:45:30,605
WE'RE WELL UNDERWAY AND HAVE 

3114
02:45:30,605 --> 02:45:34,309
DATA FOR THE CELLS AND 

3115
02:45:34,309 --> 02:45:36,578
COLLABORATING WITH YALE WE HAVE 

3116
02:45:36,578 --> 02:45:39,147
A VIRUS ON PRIMARY CD4 T CELLS 

3117
02:45:39,147 --> 02:45:41,015
AND TAKING ADVANTAGE OF THE 

3118
02:45:41,015 --> 02:45:45,920
RESTRICTION TETHERING WORKING TO

3119
02:45:45,920 --> 02:45:50,959
HELP LOOK AT THE TETHERED 

3120
02:45:50,959 --> 02:45:55,430
PARTICLES TO ENRICH TO THE CELL 

3121
02:45:55,430 --> 02:45:56,130
SURFACE.

3122
02:45:56,130 --> 02:46:01,369
WITH THIS I WILL ACKNOWLEDGE 

3123
02:46:01,369 --> 02:46:03,004
PEOPLE WHO DID THE WORK AND 

3124
02:46:03,004 --> 02:46:07,575
COLLABORATORS AND THANKS FOR THE

3125
02:46:07,575 --> 02:46:09,978
GENEROUS SUPPORT FROM NIAID AND 

3126
02:46:09,978 --> 02:46:14,716
THE IN SITU WORK IS NOT FUNDED 

3127
02:46:14,716 --> 02:46:17,018
YET.

3128
02:46:17,018 --> 02:46:23,791
THANK YOU FOR YOUR ATTENTION.

3129
02:46:23,791 --> 02:46:24,359
>> THANK YOU.

3130
02:46:24,359 --> 02:46:26,761
WHERE WE GO FOR THE QUESTIONS I 

3131
02:46:26,761 --> 02:46:28,529
WANT TO REMIND PEOPLE WE HAVE A 

3132
02:46:28,529 --> 02:46:29,964
PRESENTATION THIS AFTERNOON 

3133
02:46:29,964 --> 02:46:33,968
PLEASE BRING THE PRESENTATIONS 

3134
02:46:33,968 --> 02:46:34,669
NOW AS SOON AS THE QUESTION 

3135
02:46:34,669 --> 02:46:45,213
SESSION IS OVER WE CAN DO THAT.

3136
02:47:01,462 --> 02:47:03,298
>> GREAT WORK.

3137
02:47:03,298 --> 02:47:05,566
DO YOU HAVE ANY IDEA HOW THE B 

3138
02:47:05,566 --> 02:47:07,201
FACTORS FOR THE RECONSTRUCTION 

3139
02:47:07,201 --> 02:47:11,072
IS COMPARED BETWEEN THE IN SITU 

3140
02:47:11,072 --> 02:47:16,477
AND EX SITU SAMPLES AND HOW MUCH

3141
02:47:16,477 --> 02:47:17,979
MORE DATA DO YOU NEED IN THE 

3142
02:47:17,979 --> 02:47:24,152
CELL FOR THE RESOLUTIONS AND 

3143
02:47:24,152 --> 02:47:28,022
NOISE?

3144
02:47:28,022 --> 02:47:31,926
>> IT'S PROBABLY AT THE
LAW

3145
02:47:31,926 --> 02:47:34,962
LAWYER -- LOWER END OF THE 

3146
02:47:34,962 --> 02:47:37,965
SIGNAL TO NOISE.

3147
02:47:37,965 --> 02:47:43,338
THE 3.6 IS FROM 20 TO 30K 

3148
02:47:43,338 --> 02:47:44,172
PARTICLES GETTING TO HIGH 

3149
02:47:44,172 --> 02:47:45,973
RESOLUTION WE NEED OVER A 

3150
02:47:45,973 --> 02:47:56,284
MILLION PARTICLES.

3151
02:48:01,789 --> 02:48:03,858
>> THANK YOU.

3152
02:48:03,858 --> 02:48:07,929
SOONER OR LATER PEOPLE WORK ON 

3153
02:48:07,929 --> 02:48:09,130
THE RIBOSOME.

3154
02:48:09,130 --> 02:48:15,403
PRESUMABLY THERE'S NO PREFERRED 

3155
02:48:15,403 --> 02:48:17,071
ORIENTATIONS AND GET ISOTROPIC 

3156
02:48:17,071 --> 02:48:17,972
SETS OF IMAGES.

3157
02:48:17,972 --> 02:48:19,407
IS THAT TRUE?

3158
02:48:19,407 --> 02:48:25,046
>> I THINK SO BECAUSE IN
THE 

3159
02:48:25,046 --> 02:48:29,150
CELL THERE'S NOTHING AND YOU SEE

3160
02:48:29,150 --> 02:48:33,087
IT INSIGHT THE CELL.

3161
02:48:33,087 --> 02:48:35,056
-- INSIDE THE CELL.

3162
02:48:35,056 --> 02:48:37,925
>> A QUESTION FOR MELISSA.

3163
02:48:37,925 --> 02:48:39,260
>> HI, HARRY.

3164
02:48:39,260 --> 02:48:43,631
>> HEY.

3165
02:48:43,631 --> 02:48:51,406
I WAS WONDERING IF MX2 IS 

3166
02:48:51,406 --> 02:49:00,848
INHIBITED BY CELL CYCLE STAGE.

3167
02:49:00,848 --> 02:49:04,952
IN SOME IT CAUSES CELL CYCLE 

3168
02:49:04,952 --> 02:49:07,955
ARREST PHENOTYPE AND HAVEN'T 

3169
02:49:07,955 --> 02:49:09,290
BEEN ABLE TO DISENTANGLE THE 

3170
02:49:09,290 --> 02:49:11,192
CELL CYCLE IN YOUR SYSTEMS.

3171
02:49:11,192 --> 02:49:13,561
>> DISENTANGLES NO BUT I
CAN 

3172
02:49:13,561 --> 02:49:14,929
TELL YOU MX2 ACTIVITY AGAINST 

3173
02:49:14,929 --> 02:49:20,001
HIV IS ENHANCED WHEN YOU BLOCK 

3174
02:49:20,001 --> 02:49:21,035
CELL DIVISION SO WHICH MAKES 

3175
02:49:21,035 --> 02:49:24,071
SENSE THAT IT'S BETTER ABLE TO 

3176
02:49:24,071 --> 02:49:27,508
INHIBIT THE VIRUS WHEN THERE'S 

3177
02:49:27,508 --> 02:49:29,944
NO NEW MITOSIS HAPPENING OR 

3178
02:49:29,944 --> 02:49:32,580
PORES BEING GENERATED.

3179
02:49:32,580 --> 02:49:34,515
IT DOES HAVE ENHANCED ACTIVITY.

3180
02:49:34,515 --> 02:49:37,452
HOW THE CELL CYCLE IS PLAYING A 

3181
02:49:37,452 --> 02:49:38,286
ROLE WE DON'T KNOW BUT SHORT 

3182
02:49:38,286 --> 02:49:38,586
ANSWER, YES.

3183
02:49:38,586 --> 02:49:48,729
THANKS.

3184
02:49:49,063 --> 02:49:51,232
>> ANYMORE QUESTIONS?

3185
02:49:51,232 --> 02:49:54,335
IF NOT I'LL THANK THE SPEAKERS 

3186
02:49:54,335 --> 02:49:56,170
FOR THE MORNING'S PRESENTATIONS.

3187
02:49:56,170 --> 02:49:59,073
MAY I CALL ON THE SPEAKERS WHO
HAVE THEIR PRESENTATIONS THIS 

3188
02:49:59,073 --> 02:50:00,357
AFTERNOON TO COME UP AND LOAD UP
THE SLIDES.

3189
02:50:01,939 --> 02:50:02,807
MODERATING THIS AFTERNOON.

3190
02:50:02,807 --> 02:50:04,575
THANK YOU FOR JOINING US FOR

3191
02:50:04,575 --> 02:50:08,212
THIS LAST SESSION.

3192
02:50:08,212 --> 02:50:11,782
OUR FIRST SPEAKER IS ANDREW

3193
02:50:11,782 --> 02:50:12,883
DELAITSCH FROM CALIFORNIA

3194
02:50:12,883 --> 02:50:14,552
INSTITUTE OF TECHNOLOGY.

3195
02:50:14,552 --> 02:50:14,852
>> GREAT.

3196
02:50:14,852 --> 02:50:15,219
HELLO, EVERYONE.

3197
02:50:15,219 --> 02:50:16,320
I'D LIKE TO THANK THE ORGANIZERS

3198
02:50:16,320 --> 02:50:18,355
FOR THE OPPORTUNITY TO SHARE MY

3199
02:50:18,355 --> 02:50:20,424
WORK TODAY.

3200
02:50:20,424 --> 02:50:23,227
I'M ANDY DELAITSCH, FIFTH YEAR

3201
02:50:23,227 --> 02:50:24,628
GRADUATE STUDENT, TODAY TALKING

3202
02:50:24,628 --> 02:50:28,899
ABOUT NANOBODY WITH A COOL NAME,

3203
02:50:28,899 --> 02:50:29,133
007.

3204
02:50:29,133 --> 02:50:30,768
AND SO, THIS ANTIBODY CAME FROM

3205
02:50:30,768 --> 02:50:36,907
A COLLABORATION BETWEEN OUR LAB

3206
02:50:36,907 --> 02:50:40,711
AND KLEIN LAB IN COLOGNE,

3207
02:50:40,711 --> 02:50:41,345
GERMANY.

3208
02:50:41,345 --> 02:50:45,883
PEOPLE LIVING WITH HIV, THEY

3209
02:50:45,883 --> 02:50:47,451
SCREENED SERUM FROM 2,000 PEOPLE

3210
02:50:47,451 --> 02:50:49,753
LIVING WITH HIV, AND SCREENED

3211
02:50:49,753 --> 02:50:53,224
THE SERUM FOR ITS ABILITY TO

3212
02:50:53,224 --> 02:50:55,659
NEUTRALIZE DIVERSE VIRUSES, TAKE

3213
02:50:55,659 --> 02:50:58,496
THE TOP, ELITE NEUTRALIZERS,

3214
02:50:58,496 --> 02:51:00,998
ISOLATE SINGLE B CELLS FROM

3215
02:51:00,998 --> 02:51:01,899
THESE PEOPLE.

3216
02:51:01,899 --> 02:51:07,638
AND SO TO DO THIS THEY LOOK FOR

3217
02:51:07,638 --> 02:51:11,308
IgG POSITIVE BG 505 SINGLE B

3218
02:51:11,308 --> 02:51:14,712
CELL CLONES, FROM THE TOP ELITE

3219
02:51:14,712 --> 02:51:16,914
NEUTRALIZERS, ISOLATED HUNDREDS

3220
02:51:16,914 --> 02:51:17,448
OF MONOCLONAL ANTIBODIES,

3221
02:51:17,448 --> 02:51:18,849
SCREENED THESE FIRST ON A

3222
02:51:18,849 --> 02:51:20,718
SCREENING PANEL AND THEN ON

3223
02:51:20,718 --> 02:51:21,585
LARGER PANELS.

3224
02:51:21,585 --> 02:51:22,887
SO THE ANTIBODY THAT I'M GOING

3225
02:51:22,887 --> 02:51:25,956
TO TALK ABOUT TODAY CAME FROM

3226
02:51:25,956 --> 02:51:29,360
THIS TOP ELITE NEUTRALIZER, IT

3227
02:51:29,360 --> 02:51:39,870
LANDED IN WELL D 07 MISREAD AT

3228
02:51:44,875 --> 02:51:46,443
007, THE NAME STUCK.

3229
02:51:46,443 --> 02:51:48,846
IT APPEARS TO BE BEST IN CLASS,

3230
02:51:48,846 --> 02:51:52,583
HERE SHOWN AGAINST 119 STRAINS.

3231
02:51:52,583 --> 02:51:55,085
SO THEIR LAB TURNED TO OUR LAB

3232
02:51:55,085 --> 02:51:56,921
TO A SINGLE PARTICLE CryoEM

3233
02:51:56,921 --> 02:51:58,155
STRUCTURE OF THE ANTIBODY, I'M

3234
02:51:58,155 --> 02:51:59,323
SHOWING HERE THE TYPICAL WORK

3235
02:51:59,323 --> 02:52:03,861
FLOW THAT OUR LAB USES, AND SO

3236
02:52:03,861 --> 02:52:05,829
THIS INVOLVES MAKING FAB WITH

3237
02:52:05,829 --> 02:52:09,166
PRIMER, WE USE EXCESS OF FAB,

3238
02:52:09,166 --> 02:52:12,136
3.3 MOLES PER EVERY MOLE OF

3239
02:52:12,136 --> 02:52:12,536
TRIMER.

3240
02:52:12,536 --> 02:52:15,539
AND THEN PURIFIED SAMPLE BY SIZE

3241
02:52:15,539 --> 02:52:17,541
EXCLUSION CHROMATOGRAPHY, INDEED

3242
02:52:17,541 --> 02:52:20,444
WE SEE EXCESS UNBOUND FAB IN THE

3243
02:52:20,444 --> 02:52:21,779
SEC PROFILE.

3244
02:52:21,779 --> 02:52:24,782
WE TAKE THE COMPLEX PEAK, FREEZE

3245
02:52:24,782 --> 02:52:27,885
IT, AND THEN IMAGE ON ELECTRON

3246
02:52:27,885 --> 02:52:28,552
MICROSCOPE.

3247
02:52:28,552 --> 02:52:30,521
FROM THOUSANDS OF IMAGES LIKE

3248
02:52:30,521 --> 02:52:31,622
THIS WE'RE ABLE TO EXTRACT

3249
02:52:31,622 --> 02:52:33,157
PARTICLES AND DO DATA PROCESSING

3250
02:52:33,157 --> 02:52:36,193
AND ONE OF THE BEAUTIFUL THINGS

3251
02:52:36,193 --> 02:52:38,095
ABOUT E.M., IT DEALS WITH

3252
02:52:38,095 --> 02:52:39,830
HETEROGENEITY QUITE NICELY SO

3253
02:52:39,830 --> 02:52:42,066
FROM A SINGLE DATASET WE GET NOT

3254
02:52:42,066 --> 02:52:44,435
ONE STRUCTURE BUT FOUR

3255
02:52:44,435 --> 02:52:45,970
STRUCTURES, HERE YOU CAN SEE

3256
02:52:45,970 --> 02:52:48,105
UNBOUND, ONE BOUND, TWO BOUND,

3257
02:52:48,105 --> 02:52:50,908
THREE BOUND STRUCTURES.

3258
02:52:50,908 --> 02:52:53,244
SO WHILE THIS UNUSUAL

3259
02:52:53,244 --> 02:52:55,145
STOICHIOMETRY IS THE FOCUS, I

3260
02:52:55,145 --> 02:52:57,014
WANT TO HIGHLIGHT SOME FEATURES

3261
02:52:57,014 --> 02:52:59,083
OF THE EPITOPE OF THIS ANTIBODY,

3262
02:52:59,083 --> 02:53:02,486
LOOKING SPECIFICALLY AT THE ONE

3263
02:53:02,486 --> 02:53:03,254
BOUND STRUCTURE.

3264
02:53:03,254 --> 02:53:10,628
AND SO AS I MENTIONED IT'S A V3

3265
02:53:10,628 --> 02:53:13,197
ANTIBODY AT THE APEX, THEY BIND

3266
02:53:13,197 --> 02:53:14,431
WITH DIFFERENT ANGLES OF

3267
02:53:14,431 --> 02:53:15,065
APPROACHES.

3268
02:53:15,065 --> 02:53:18,235
AS YOU CAN SEE ON THE LEFT 007

3269
02:53:18,235 --> 02:53:21,505
SEEMS TO BIND TO ONE SIDE

3270
02:53:21,505 --> 02:53:25,009
RELATIVE TO OTHER HIV V3

3271
02:53:25,009 --> 02:53:27,778
bNAbS, IT DOES SO, HOISTED UP

3272
02:53:27,778 --> 02:53:32,316
IN BETWEEN THESE N 156 AND M 301

3273
02:53:32,316 --> 02:53:36,186
GLYCANS, DOES NOT COMBER ACT

3274
02:53:36,186 --> 02:53:41,191
WITH N332, AND HAS 22 AMINO

3275
02:53:41,191 --> 02:53:43,227
ACID-LONG CDRH 3 WHICH MAKES

3276
02:53:43,227 --> 02:53:48,565
CONTACT WITH COULD BE SERVED

3277
02:53:48,565 --> 02:53:49,833
GDIR MOTIF ON V3.

3278
02:53:49,833 --> 02:53:55,139
THE MOST HAVING THING TO ME WE

3279
02:53:55,139 --> 02:53:55,906
OBSERVED BIND BUYING ELECTRON

3280
02:53:55,906 --> 02:53:58,609
MICROSCOPY, AND SO THIS IS

3281
02:53:58,609 --> 02:54:00,277
REALLY UNUSUAL BECAUSE THIS IS A

3282
02:54:00,277 --> 02:54:06,150
VERY POTENT ANTIBODY, IT WAS

3283
02:54:06,150 --> 02:54:09,953
POTENT AGAINST 505, WE THINK OF

3284
02:54:09,953 --> 02:54:11,388
NEUTRALIZATION POTENCY AS

3285
02:54:11,388 --> 02:54:13,223
CORRELATED WITH BINDING, AND SO

3286
02:54:13,223 --> 02:54:14,792
BY EM LOOKS LIKE THIS ANTIBODY

3287
02:54:14,792 --> 02:54:15,759
IS NOT BINDING WELL.

3288
02:54:15,759 --> 02:54:19,129
YOU CAN SEE THE DISTRIBUTION OF

3289
02:54:19,129 --> 02:54:20,798
PARTICLES HERE ON THE RIGHT.

3290
02:54:20,798 --> 02:54:22,766
AND SO WHAT IS GOING ON?

3291
02:54:22,766 --> 02:54:25,602
THIS WAS THE QUESTION.

3292
02:54:25,602 --> 02:54:27,805
AND WE REALLY MADE NO PROGRESS

3293
02:54:27,805 --> 02:54:29,206
FOR WELL OVER A YEAR UNTIL ONE

3294
02:54:29,206 --> 02:54:32,609
DAY I HAD SORT OF A CRAZY

3295
02:54:32,609 --> 02:54:34,912
HYPOTHESIS, WHICH IS THAT

3296
02:54:34,912 --> 02:54:38,749
PERHAPS 007 IgG IS BINDING

3297
02:54:38,749 --> 02:54:40,284
BIVALENTLY TO A SINGLE OM.

3298
02:54:40,284 --> 02:54:42,653
IN THE TOP RIGHT YOU CAN SEE THE

3299
02:54:42,653 --> 02:54:45,522
FAB USED FOR THE STRUCTURAL

3300
02:54:45,522 --> 02:54:47,291
DETERMINATION BUT THE IgG,

3301
02:54:47,291 --> 02:54:52,096
BIOLOGICALLY RELEVANT FORM, WAS

3302
02:54:52,096 --> 02:54:52,830
USED FOR NEUTRALIZATION ASSAY ON

3303
02:54:52,830 --> 02:54:53,630
THE RIGHT.

3304
02:54:53,630 --> 02:54:55,132
THIS IS A CRAZY HYPOTHESIS, FOR

3305
02:54:55,132 --> 02:54:56,266
TWO REASONS.

3306
02:54:56,266 --> 02:54:59,303
ONE IS THAT TO MY KNOWLEDGE THIS

3307
02:54:59,303 --> 02:55:09,847
HAS NEVER BEEN SHOWN FOR ANY HIV

3308
02:55:10,981 --> 02:55:14,084
ANTIBODIES BEFORE, SECONDLY THE

3309
02:55:14,084 --> 02:55:17,488
WAY PEOPLE INFERRED IS LOOKING

3310
02:55:17,488 --> 02:55:19,123
AT HEAVY CHAINS.

3311
02:55:19,123 --> 02:55:21,158
C TERM NIGH ARE THE SHOULDERS,

3312
02:55:21,158 --> 02:55:23,727
THEY HAVE TO BE WITHIN A CERTAIN

3313
02:55:23,727 --> 02:55:27,364
DISTANCE, SHOULDER WIDTH APART

3314
02:55:27,364 --> 02:55:33,170
FOR ANTIBODY, ABOUT 50 OR 60

3315
02:55:33,170 --> 02:55:33,804
ANGSTROMS.

3316
02:55:33,804 --> 02:55:35,139
IN MY TWO BOUND STRUCTURE YOU

3317
02:55:35,139 --> 02:55:42,880
CAN SEE THE C TERMINI ARE TOO

3318
02:55:42,880 --> 02:55:43,881
FAR APART.

3319
02:55:43,881 --> 02:55:47,484
THIS IS A DYNAMIC MACHINE AND AS

3320
02:55:47,484 --> 02:55:49,453
MANY PEOPLE HERE KNOW, THERE

3321
02:55:49,453 --> 02:55:54,758
HAVE BEEN OPEN STATES OF OHMs

3322
02:55:54,758 --> 02:55:56,593
SOLVED, HERE OCCLUDED OPEN OHMS,

3323
02:55:56,593 --> 02:55:59,763
WHEN I TOOK MY STRUCTURE, A

3324
02:55:59,763 --> 02:56:01,865
CLOSED OMM, TOOK THE FAB

3325
02:56:01,865 --> 02:56:07,004
RELATIVE TO THE GP120 AND DOCKED

3326
02:56:07,004 --> 02:56:11,241
TO VARIOUS OPEN FORMS C TERMINI

3327
02:56:11,241 --> 02:56:12,142
COME IN CLOSE PROXIMITY, COOL

3328
02:56:12,142 --> 02:56:13,710
BUT STILL A MODEL, NO EVIDENCE

3329
02:56:13,710 --> 02:56:14,344
FOR THIS.

3330
02:56:14,344 --> 02:56:16,513
BUT WHAT IT DID DO IS PROMPTED

3331
02:56:16,513 --> 02:56:19,249
US TO RUN ANOTHER EXPERIMENT,

3332
02:56:19,249 --> 02:56:28,258
WHICH IS SOAP  -- SHOWN HERE,

3333
02:56:28,258 --> 02:56:29,326
NEUTRALIZATION ASSAY, COMPARING

3334
02:56:29,326 --> 02:56:31,628
HOW POTENT IS FAB RELATIVE TO

3335
02:56:31,628 --> 02:56:31,962
IgG.

3336
02:56:31,962 --> 02:56:33,063
THE IgG IS ABOUT THREE TIMES

3337
02:56:33,063 --> 02:56:39,937
THE SIZE OF THE FAB, TO CONTROL

3338
02:56:39,937 --> 02:56:42,306
FOR THE STERICS, ONE ARM IS

3339
02:56:42,306 --> 02:56:44,441
NON-HIV BINDING FAB, AND SO

3340
02:56:44,441 --> 02:56:48,812
THAT'S A BETTER CONTROLLED

3341
02:56:48,812 --> 02:56:53,083
COMPARISON THIS IS SHOWN FOR ONE

3342
02:56:53,083 --> 02:56:54,818
EXAMPLE PSEUDOVIRUS, MOVING FROM

3343
02:56:54,818 --> 02:56:57,654
THE FAB TO BI-SPECIFIC WE GET A

3344
02:56:57,654 --> 02:57:01,091
20-FOLD INCREASE IN POTENCY,

3345
02:57:01,091 --> 02:57:03,694
INDICATING THERE'S SOME STERIC

3346
02:57:03,694 --> 02:57:07,631
COMPONENT, AND FROM BI-SPECIFIC

3347
02:57:07,631 --> 02:57:10,367
TO NATIVE IgG WE GET

3348
02:57:10,367 --> 02:57:11,468
ADDITIONAL 250-FOLD INCREASE IN

3349
02:57:11,468 --> 02:57:13,203
POTENCY FOR THIS SPECIFIC

3350
02:57:13,203 --> 02:57:13,437
STRAIN.

3351
02:57:13,437 --> 02:57:16,607
SO, TOGETHER THAT MEANS THAT THE

3352
02:57:16,607 --> 02:57:17,908
IgG IS ABOUT 5,000 TIMES MORE

3353
02:57:17,908 --> 02:57:20,878
POTENT THAN FAB FOR THIS STRAIN.

3354
02:57:20,878 --> 02:57:24,081
AND SO THAT WAS PRETTY

3355
02:57:24,081 --> 02:57:25,582
UNEXPECTED AND COOL.

3356
02:57:25,582 --> 02:57:27,651
AND SO WE THOUGHT THIS WAS GOING

3357
02:57:27,651 --> 02:57:28,986
TO BE SOMEWHAT STRAIGHTFORWARD.

3358
02:57:28,986 --> 02:57:31,522
LET'S DO A SINGLE PARTICLE EM

3359
02:57:31,522 --> 02:57:34,458
STRUCTURE AND SEE IF WE SEE THAT

3360
02:57:34,458 --> 02:57:35,759
STRUCTURE THAT WE EXPECTED.

3361
02:57:35,759 --> 02:57:42,599
AND SO FOR THIS I SIMPLIFIED THE

3362
02:57:42,599 --> 02:57:48,805
WORKFLOW, MIXED EQUI MOLAR 007

3363
02:57:48,805 --> 02:57:50,774
WITH THE 505 SOSIP, WE SEE

3364
02:57:50,774 --> 02:57:52,209
SOMETHING THAT LOOKS LIKE THIS,

3365
02:57:52,209 --> 02:57:55,178
TO OUR SURPRISE.

3366
02:57:55,178 --> 02:57:57,347
THE EQUILIBRIUM IS SHIFTED TO

3367
02:57:57,347 --> 02:57:58,148
THE WEIRD TRIMER-DIMER

3368
02:57:58,148 --> 02:57:59,016
STRUCTURE, FOR THOSE FAMILIAR

3369
02:57:59,016 --> 02:58:00,651
WITH EM THIS IS A GOOD REMINDER

3370
02:58:00,651 --> 02:58:02,719
THAT THINGS THAT ARE FLEXIBLE IN

3371
02:58:02,719 --> 02:58:06,523
EM SUCH AS UNBOUND FABS OR SD

3372
02:58:06,523 --> 02:58:07,624
ARE NOT OBSERVED BECAUSE THIS IS

3373
02:58:07,624 --> 02:58:11,562
AT ITS CORE AN AVERAGING

3374
02:58:11,562 --> 02:58:15,365
TECHNIQUE, STRUCTURES CAN BE

3375
02:58:15,365 --> 02:58:16,300
INTERPRETED MORE LIKE THAT,

3376
02:58:16,300 --> 02:58:17,701
IgG AROUND THE FABs, BUT ON

3377
02:58:17,701 --> 02:58:20,003
THE RIGHT THE STRUCTURE IS

3378
02:58:20,003 --> 02:58:22,272
REALLY WEIRD. 

3379
02:58:22,272 --> 02:58:24,808
AND SO WE ASKED THE QUESTION CAN

3380
02:58:24,808 --> 02:58:27,210
THIS STRUCTURE ACTUALLY EXPLAIN

3381
02:58:27,210 --> 02:58:30,881
THIS INCREASED NEUTRALIZATION

3382
02:58:30,881 --> 02:58:31,682
POTENCY OF THE IgG?

3383
02:58:31,682 --> 02:58:33,350
THE WAY WE THINK OF THIS THE

3384
02:58:33,350 --> 02:58:35,852
POTENTIAL MECHANISM WOULD BE

3385
02:58:35,852 --> 02:58:38,789
THAT YOU HAVE ONE ON ONE VIRUS,

3386
02:58:38,789 --> 02:58:40,591
ANOTHER ON A DIFFERENT VIRUS,

3387
02:58:40,591 --> 02:58:42,492
AND YOU WOULD HAVE VIRAL

3388
02:58:42,492 --> 02:58:46,129
AGGREGATION AS A MECHANISM OF

3389
02:58:46,129 --> 02:58:46,897
NEUTRALIZATION, FORMER THIS

3390
02:58:46,897 --> 02:58:49,166
TRIMER-DIMER STRUCTURE IN

3391
02:58:49,166 --> 02:58:51,134
BETWEEN VIRIONS.

3392
02:58:51,134 --> 02:58:54,638
THERE'S ALSO OUR ORIGINAL

3393
02:58:54,638 --> 02:58:55,739
HYPOTHESIS OF INTRASPIKE

3394
02:58:55,739 --> 02:58:57,608
CROSS-LINKING TO SINGLE OPEN OM

3395
02:58:57,608 --> 02:59:00,243
OR BINDING TO ADJACENT TO SPIKES

3396
02:59:00,243 --> 02:59:02,546
ON A SINGLE VIRUS.

3397
02:59:02,546 --> 02:59:05,182
AND SO WE WANTED TO NAIL THIS

3398
02:59:05,182 --> 02:59:07,084
HOME, AND FIGURE OUT WHAT WAS

3399
02:59:07,084 --> 02:59:08,852
GOING ON WITHIN OUR PSEUDOVIRUS

3400
02:59:08,852 --> 02:59:10,420
ASSAY, AND SO WHAT WE WOULD LIKE

3401
02:59:10,420 --> 02:59:14,758
TO DO IS IMAGE THE 007 IgG IN

3402
02:59:14,758 --> 02:59:20,330
COMPLEX WITH OTHER VIRUS OR

3403
02:59:20,330 --> 02:59:22,466
RATHER ENVELOPE VIRUS-LIKE

3404
02:59:22,466 --> 02:59:22,733
PARTICLES.

3405
02:59:22,733 --> 02:59:23,367
OKAY.

3406
02:59:23,367 --> 02:59:25,802
AND SO TO DO THIS WE UTILIZED

3407
02:59:25,802 --> 02:59:28,605
TECHNOLOGY THAT WAS PREVIOUSLY

3408
02:59:28,605 --> 02:59:31,908
DEVELOPED IN OUR LAB, EABR

3409
02:59:31,908 --> 02:59:33,343
TECHNOLOGY, THE MAIN PURPOSE AT

3410
02:59:33,343 --> 02:59:34,945
THE TIME WAS AS A VACCINE

3411
02:59:34,945 --> 02:59:36,913
PLATFORM, BUT I WANT TO MAKE THE

3412
02:59:36,913 --> 02:59:39,650
CASE IT'S USEFUL FOR STRUCTURAL

3413
02:59:39,650 --> 02:59:42,586
BIOLOGY AND SO THE IDEA HERE WE

3414
02:59:42,586 --> 02:59:46,556
HAVE A GENE SHOWN IN TOP LEFT,

3415
02:59:46,556 --> 02:59:51,194
FULL LENGTH HIV OMs, SWITCH TO

3416
02:59:51,194 --> 02:59:56,566
ENDOSIGH THOSE IS PREVENTING

3417
02:59:56,566 --> 02:59:58,068
MOTIF FOLLOWED BY EABR SEQUENCE.

3418
02:59:58,068 --> 03:00:03,840
THAT LEADS TO THE FORMATION OF

3419
03:00:03,840 --> 03:00:06,610
THESE VIRUS-LIKE PARTICLES.

3420
03:00:06,610 --> 03:00:07,177
WITH MEMBRANE PRESENT.

3421
03:00:07,177 --> 03:00:09,946
THIS IS A REALLY ADVANTAGEOUS

3422
03:00:09,946 --> 03:00:13,717
METHOD, BECAUSE RELATIVE TO

3423
03:00:13,717 --> 03:00:15,252
WORKING WITH VIRUSES, SAFE AND

3424
03:00:15,252 --> 03:00:16,653
EASY TO WORK WITH, VERSATILE,

3425
03:00:16,653 --> 03:00:17,654
ANYTHING YOU CAN CLONE YOU CAN

3426
03:00:17,654 --> 03:00:22,259
TEST TO SEE WHETHER YOU GET

3427
03:00:22,259 --> 03:00:23,794
EXPRESSION, NOT LIMITED TO A FEW

3428
03:00:23,794 --> 03:00:29,032
VIRAL STRAINS AND RELATIVE TO

3429
03:00:29,032 --> 03:00:31,334
SOSIPs, STRESSING FULL LENGTH

3430
03:00:31,334 --> 03:00:33,737
OMS WITH A MEMBRANE.

3431
03:00:33,737 --> 03:00:35,272
THE WORKFLOW IS PRETTY

3432
03:00:35,272 --> 03:00:36,039
STRAIGHTFORWARD.

3433
03:00:36,039 --> 03:00:38,675
I'M NOT GOING INTO DETAIL BUT

3434
03:00:38,675 --> 03:00:44,548
WE'VE PURIFIED ON A SUCROSE

3435
03:00:44,548 --> 03:00:46,216
GRADIENT, DIALIZED, LOOKED ON

3436
03:00:46,216 --> 03:00:46,650
MICROSCOPE.

3437
03:00:46,650 --> 03:00:50,387
THESE THINGS LOOK REALLY

3438
03:00:50,387 --> 03:00:50,921
BEAUTIFUL.

3439
03:00:50,921 --> 03:00:53,990
RELATIVE TO ACTUAL VIRUSES

3440
03:00:53,990 --> 03:00:56,693
SPARSELY COATED IN OMS, YOU CAN

3441
03:00:56,693 --> 03:00:58,795
SEE A LOT PRESENT.

3442
03:00:58,795 --> 03:01:04,301
WE'RE GETTING A NUMBER OF OMS,

3443
03:01:04,301 --> 03:01:05,402
RELATIVE POSITIONING AND TILT

3444
03:01:05,402 --> 03:01:06,503
RELATIVE TO MEMBRANE.

3445
03:01:06,503 --> 03:01:07,604
ONE THING NOT IMMEDIATELY

3446
03:01:07,604 --> 03:01:09,639
OBVIOUS YOU CAN PERFORM SINGLE

3447
03:01:09,639 --> 03:01:12,042
PARTICLE ON SIDE VIEW OMS ON

3448
03:01:12,042 --> 03:01:13,944
PERIMETER OF THE VIRUSES SO WE

3449
03:01:13,944 --> 03:01:18,548
DID THIS, AND WE'VE GOTTEN

3450
03:01:18,548 --> 03:01:19,316
ROUGHLY 7 OR 8-ANGSTROM

3451
03:01:19,316 --> 03:01:20,183
STRUCTURE OF THIS.

3452
03:01:20,183 --> 03:01:21,384
THAT'S WHERE WE'RE AT NOW.

3453
03:01:21,384 --> 03:01:23,487
SO I'LL LEAVE YOU WITH A

3454
03:01:23,487 --> 03:01:24,988
CLIFFHANGER OF THE NEXT THING WE

3455
03:01:24,988 --> 03:01:27,724
HAVE TO DO, WE WANT -- NOW THAT

3456
03:01:27,724 --> 03:01:30,594
WE'VE ESTABLISHED WE CAN USE

3457
03:01:30,594 --> 03:01:32,596
EVLPs, WE WANT TO COMPLEX THEM

3458
03:01:32,596 --> 03:01:34,064
WITH ANTIBODY OF INTEREST.

3459
03:01:34,064 --> 03:01:41,538
FUTURE DIRECTIONS TO DO THIS,

3460
03:01:41,538 --> 03:01:43,240
THIS IMAGE OF THE STABILIZED

3461
03:01:43,240 --> 03:01:44,274
CONSTRUCT, WE WANT TO SEE IF WE

3462
03:01:44,274 --> 03:01:48,512
CAN ALSO GET A NATIVE

3463
03:01:48,512 --> 03:01:49,045
NON-STABILIZED CONSTRUCT.

3464
03:01:49,045 --> 03:01:52,816
FINALLY I DIDN'T GO INTO IT BUT

3465
03:01:52,816 --> 03:01:54,384
AVIDITY IS A CONSEQUENCE, WE

3466
03:01:54,384 --> 03:01:57,387
WANT TO LOOK FURTHER INTO

3467
03:01:57,387 --> 03:02:01,191
BINDING KINETICS OF 007 WITH

3468
03:02:01,191 --> 03:02:02,425
DIFFERENT OMS.

3469
03:02:02,425 --> 03:02:04,795
I A TON OF PEOPLE TO THANK,

3470
03:02:04,795 --> 03:02:09,199
PAMELA, ADVISER, PEOPLE IN THE

3471
03:02:09,199 --> 03:02:10,400
LAB, WONDERFUL COLLABORATORS,

3472
03:02:10,400 --> 03:02:13,570
AND POSTDOC IN THE LAB, AND THEN

3473
03:02:13,570 --> 03:02:16,206
VARIOUS PEOPLE WHO HELPED

3474
03:02:16,206 --> 03:02:17,107
IMMEDIATE WITH EM, FUNDING

3475
03:02:17,107 --> 03:02:19,576
SOURCES, HAPPY TO TAKE

3476
03:02:19,576 --> 03:02:19,843
QUESTIONS.

3477
03:02:19,843 --> 03:02:29,352
[APPLAUSE] 

3478
03:02:29,352 --> 03:02:30,453
>> THANK YOU, ANDREW.

3479
03:02:30,453 --> 03:02:31,988
ANY QUESTIONS?

3480
03:02:31,988 --> 03:02:34,925
>> I WAS ABOUT TO ASK
ABOUT THE

3481
03:02:34,925 --> 03:02:38,328
KINETIC RATES UNTIL I SAW THE

3482
03:02:38,328 --> 03:02:39,663
LAST SLIDE.

3483
03:02:39,663 --> 03:02:40,530
>> YEAH, A REALLY
INTERESTING

3484
03:02:40,530 --> 03:02:47,437
THING WHEN YOU LOOK AT THESE

3485
03:02:47,437 --> 03:02:49,072
NEUTRALIZATION CURVES, HERE

3486
03:02:49,072 --> 03:02:54,344
STERIC COMPONENT, WE ALWAYS SEE

3487
03:02:54,344 --> 03:02:58,682
THIS, BUT  THE AVIDITY COMPONENT

3488
03:02:58,682 --> 03:03:02,419
VARIOUS FROM A FEW TO FEW

3489
03:03:02,419 --> 03:03:02,652
HUNDRED.

3490
03:03:02,652 --> 03:03:05,255
>> DO YOU KNOW WHAT THE
SPACING

3491
03:03:05,255 --> 03:03:08,592
DISTANCE IS BETWEEN THE OMS?

3492
03:03:08,592 --> 03:03:11,394
>> YOU CAN JUST SEE
VISUALLY A

3493
03:03:11,394 --> 03:03:13,496
LOT OF THE EVLPs ARE DENSELY

3494
03:03:13,496 --> 03:03:15,432
COATED, SPIKES ALL OVER.

3495
03:03:15,432 --> 03:03:16,900
YEAH, I'D LIKE TO DO TOMOGRAPHY

3496
03:03:16,900 --> 03:03:18,869
TO FURTHER LOOK INTO THAT.

3497
03:03:18,869 --> 03:03:20,604
>> THANK YOU.

3498
03:03:20,604 --> 03:03:22,239
>> HI.

3499
03:03:22,239 --> 03:03:24,941
CAN YOU JUST EXPAND ON WHICH

3500
03:03:24,941 --> 03:03:26,176
ESCORTS SEEM TO DO A BETTER JOB

3501
03:03:26,176 --> 03:03:29,012
AND WHY YOU THINK THAT MIGHT BE

3502
03:03:29,012 --> 03:03:29,980
HAPPENING?

3503
03:03:29,980 --> 03:03:30,981
>> ON WHICH ESCORT?

3504
03:03:30,981 --> 03:03:32,048
EXCUSE ME?

3505
03:03:32,048 --> 03:03:36,019
YEAH, SO I DIDN'T REALLY TEST

3506
03:03:36,019 --> 03:03:40,590
ANY DIFFERENT LIKE ESCORT

3507
03:03:40,590 --> 03:03:40,857
MACHINERY.

3508
03:03:40,857 --> 03:03:42,559
WE USED THIS EABR SEQUENCE

3509
03:03:42,559 --> 03:03:43,693
PREVIOUSLY PUBLISHED, WENT

3510
03:03:43,693 --> 03:03:45,528
FORWARD AND IT WORKED, SO WE

3511
03:03:45,528 --> 03:03:46,730
HAVEN'T DONE ANY TROUBLESHOOTING

3512
03:03:46,730 --> 03:03:49,833
SO I DON'T HAVE MUCH.

3513
03:03:49,833 --> 03:03:53,203
>> OKAY, THANKS.

3514
03:03:53,203 --> 03:03:56,673
>> THANK YOU AGAIN, ANDY. 

3515
03:03:56,673 --> 03:03:57,040
>> THANK YOU.

3516
03:03:57,040 --> 03:04:04,347
[APPLAUSE] 

3517
03:04:04,347 --> 03:04:12,989
>> NEXT SPEAKER IS
SHARIDAN

3518
03:04:12,989 --> 03:04:17,360
BROWN FROM SCRIPPS RESEARCH. 

3519
03:04:17,360 --> 03:04:20,730
WE'LL GIVE HER A SECOND.

3520
03:04:20,730 --> 03:04:21,698
>> I'M SHARIDAN, GRADUATE

3521
03:04:21,698 --> 03:04:23,166
STUDENT IN ANDREW WARD'S LAB AT

3522
03:04:23,166 --> 03:04:25,669
SCRIPPS, TALKING ABOUT MY WORK

3523
03:04:25,669 --> 03:04:28,338
ON ANTI-IMMUNE COMPLEX

3524
03:04:28,338 --> 03:04:30,040
ANTIBODIES ELICITED AGAINST HIV

3525
03:04:30,040 --> 03:04:30,206
OMS.

3526
03:04:30,206 --> 03:04:32,142
FIRST I WANT TO TOUCH ON SOME OF

3527
03:04:32,142 --> 03:04:34,377
THE CORE GOALS OF RATIONAL

3528
03:04:34,377 --> 03:04:35,111
VACCINE DESIGN.

3529
03:04:35,111 --> 03:04:39,182
PRETTY MUCH WE WANT TO ANALYZE

3530
03:04:39,182 --> 03:04:41,451
ANTIBODY RESPONSES, AND

3531
03:04:41,451 --> 03:04:42,552
CHARACTERIZE DIFFERENT

3532
03:04:42,552 --> 03:04:43,453
NEUTRALIZATION CAPABILITIES OF

3533
03:04:43,453 --> 03:04:53,964
ANTIBODIES TO GO BACK INTO OUR

3534
03:04:57,267 --> 03:05:06,743
IMMUNOGENS, PRODUCING GERMLINE

3535
03:05:06,743 --> 03:05:08,378
TARGETING IMMUNEEGENS, I'LL TAKE

3536
03:05:08,378 --> 03:05:09,679
YOU THROUGH THE PIPELINE.

3537
03:05:09,679 --> 03:05:13,750
WE'RE ABLE TO TAKE IMMUNIZED

3538
03:05:13,750 --> 03:05:15,819
SERA, ISOLATE IgG, DIGEST INTO

3539
03:05:15,819 --> 03:05:22,325
FAB WHICH WE CAN RECOMPLEX WITH

3540
03:05:22,325 --> 03:05:23,693
ANTIGEN, PUT THAT PUT THAT ON

3541
03:05:23,693 --> 03:05:26,596
CryoEM OR NEGATIVE STAINING

3542
03:05:26,596 --> 03:05:26,863
GRIDS.

3543
03:05:26,863 --> 03:05:29,299
THROUGH SERIES OF CLASSIFICATION

3544
03:05:29,299 --> 03:05:31,034
STEPS WE CAN ISOLATE HIGH

3545
03:05:31,034 --> 03:05:35,038
RESOLUTION MODELS OF OUR ANTIGEN

3546
03:05:35,038 --> 03:05:37,407
BOUND TO POLYCLONAL ANTIBODIES.

3547
03:05:37,407 --> 03:05:38,208
THIS COMPLEMENTS RATIONAL

3548
03:05:38,208 --> 03:05:39,476
VACCINE DESIGN TECHNIQUES.

3549
03:05:39,476 --> 03:05:43,646
WE'VE USED IT TO HELP PRODUCE

3550
03:05:43,646 --> 03:05:44,314
STABLE AND SOLUBLE

3551
03:05:44,314 --> 03:05:46,716
GLYCOPROTEINS, THAT COULD BE

3552
03:05:46,716 --> 03:05:48,985
USED AS IMMUNOGENS.

3553
03:05:48,985 --> 03:05:50,186
I WANT TO HIGHLIGHT WORK BY

3554
03:05:50,186 --> 03:05:52,722
OTHER PEOPLE IN OUR LAB BECAUSE

3555
03:05:52,722 --> 03:05:59,496
I THINK IT SHOWCASES EXACTLY HOW

3556
03:05:59,496 --> 03:06:00,263
POWERFUL EMPEM.

3557
03:06:00,263 --> 03:06:03,333
SHE WAS ABLE TO PRODUCE A

3558
03:06:03,333 --> 03:06:10,240
DATASET CONTAINING 20 UNIQUE

3559
03:06:10,240 --> 03:06:10,840
POLYCLONAL ANTIBODIES, RANGING

3560
03:06:10,840 --> 03:06:18,214
FROM 3 TO 3 1/2 ANGSTROMS,

3561
03:06:18,214 --> 03:06:25,822
TARGET SIX THREE ON THE ENVELOPE

3562
03:06:25,822 --> 03:06:28,858
SEES BEYOND INCLUDING MALARIA,

3563
03:06:28,858 --> 03:06:30,627
GLYCOPROTEIN, INFLUENZA VIRUS,

3564
03:06:30,627 --> 03:06:35,398
CORONAVIRUS SPIKE, AND HEPATITIS

3565
03:06:35,398 --> 03:06:35,632
C VIRUS.

3566
03:06:35,632 --> 03:06:37,734
MY WORK, I WANT TO FIRST

3567
03:06:37,734 --> 03:06:42,205
INTRODUCE THE CONCEPT OF

3568
03:06:42,205 --> 03:06:42,872
ANTI-IDIOTIPPIC ANTIBODIES, THAT

3569
03:06:42,872 --> 03:06:49,012
TARGET AND BIND TO PRIMARY

3570
03:06:49,012 --> 03:06:51,514
ANTIBODIES, IN THREE CLASSES,

3571
03:06:51,514 --> 03:06:54,551
AB2 BIND TO PRIMARY ANTIBODIES.

3572
03:06:54,551 --> 03:06:57,420
AB2 BETA BIND INTO THE CDR

3573
03:06:57,420 --> 03:06:59,823
REGION OF THE PRIMARY ANTIBODY.

3574
03:06:59,823 --> 03:07:03,893
AND THIS BLOCKS ANTIGEN BINDING.

3575
03:07:03,893 --> 03:07:06,529
AND THEN AB2 GAMMA ANTIBODIES

3576
03:07:06,529 --> 03:07:12,202
BIND ADJACENT TO CDR BINDING

3577
03:07:12,202 --> 03:07:17,173
SITE BLOCK ANTIGEN BINDING BUT

3578
03:07:17,173 --> 03:07:20,076
ARE NOT INTERNAL IMAGE

3579
03:07:20,076 --> 03:07:20,543
ANTIBODIES.

3580
03:07:20,543 --> 03:07:24,848
WE CAME ACROSS A SERIES OF

3581
03:07:24,848 --> 03:07:26,216
ANTIBODIES, ANTI-IMMUNE COMPLEX

3582
03:07:26,216 --> 03:07:27,250
ANTIBODIES, IN FOUR CLASSES.

3583
03:07:27,250 --> 03:07:29,953
I'LL WALK YOU THROUGH EACH OF

3584
03:07:29,953 --> 03:07:31,554
THOSE IN DETAIL.

3585
03:07:31,554 --> 03:07:34,190
OUR FIRST EXAMPLE IS CLASS 1

3586
03:07:34,190 --> 03:07:35,191
ANTI-IMMUNE COMPLEX ANTIBODIES,

3587
03:07:35,191 --> 03:07:38,595
AND THESE ARE ANTIBODIES THAT

3588
03:07:38,595 --> 03:07:41,664
HAVE DISTINCT ANTIGEN EPITOPES

3589
03:07:41,664 --> 03:07:43,199
BUT MAKE MINOR NON-SPECIFIC

3590
03:07:43,199 --> 03:07:44,400
CONTACTS ALONG THE FRAMEWORK

3591
03:07:44,400 --> 03:07:44,634
REGIONS.

3592
03:07:44,634 --> 03:07:52,041
IN THIS EXAMPLE WE HAVE A BASE

3593
03:07:52,041 --> 03:07:54,144
ANTIBODY, N625 FAB MAKING MINOR

3594
03:07:54,144 --> 03:07:54,911
CONTACTS.

3595
03:07:54,911 --> 03:07:57,180
SAME ANIMAL, SAME BASE EPITOPE,

3596
03:07:57,180 --> 03:08:03,920
BUT WE ALSO HAVE AN ADDITIONAL N

3597
03:08:03,920 --> 03:08:07,257
66 ANTIBODY BINDING, WE THINK

3598
03:08:07,257 --> 03:08:08,825
ONE BINDS, HELPS STABILIZE

3599
03:08:08,825 --> 03:08:10,426
EPITOPE, ALLOWS OTHER ANTIBODY

3600
03:08:10,426 --> 03:08:15,398
TO COME IN AND BIND.

3601
03:08:15,398 --> 03:08:17,333
THESE CLASS 2 ANTI-IMMUNE

3602
03:08:17,333 --> 03:08:19,235
COMPLEX ARE EPITOPE OF ANTIGEN

3603
03:08:19,235 --> 03:08:21,838
AND ANTIBODY, SO IN THIS CASE

3604
03:08:21,838 --> 03:08:23,039
THE ANTI-IMMUNE COMPLEX ANTIBODY

3605
03:08:23,039 --> 03:08:27,310
IN GREEN IS BINDING TO A GLYCAN

3606
03:08:27,310 --> 03:08:28,745
ON OUR SOSIP ANTIGEN, ALSO

3607
03:08:28,745 --> 03:08:35,185
BINDING TO THE FRAMEWORK REGIONS

3608
03:08:35,185 --> 03:08:38,354
OF THIS N 241 FAB.

3609
03:08:38,354 --> 03:08:42,058
ANOTHER EXAMPLE BETWEEN THE BLUE

3610
03:08:42,058 --> 03:08:49,732
AND B1B 3 FAB CONTACTING

3611
03:08:49,732 --> 03:08:52,602
FRAMEWORK REGIONS BUT ALSO THIS

3612
03:08:52,602 --> 03:08:53,770
138 GLYCAN.

3613
03:08:53,770 --> 03:08:59,375
CLASS III ARE YOUR

3614
03:08:59,375 --> 03:09:02,011
QUINTESSENTIAL AB2 ALPHA

3615
03:09:02,011 --> 03:09:02,612
IDIOTYPIC ANTIBODIES, COMPOSED

3616
03:09:02,612 --> 03:09:04,214
OF ANTIBODY, DOES NOT MAKE

3617
03:09:04,214 --> 03:09:06,282
CONTACT WITH THE ANTIGEN,

3618
03:09:06,282 --> 03:09:07,717
THERE'S EXTENSIVE CONTACTS WITH

3619
03:09:07,717 --> 03:09:12,088
FRAMEWORK REGION 1 AND FRAMEWORK

3620
03:09:12,088 --> 03:09:18,995
REGION 3 OF THIS V2 FAB.

3621
03:09:18,995 --> 03:09:23,032
PERHAPS CLASS 4 ANTIBODY HAS

3622
03:09:23,032 --> 03:09:24,334
EPITOPE OF TWO DISTINCT

3623
03:09:24,334 --> 03:09:27,437
ANTIBODIES, THIS IS BINDING TO

3624
03:09:27,437 --> 03:09:32,875
BOTH THIS V2 FAB IN BLUE AND GP1

3625
03:09:32,875 --> 03:09:34,310
INTERFACE IN RED.

3626
03:09:34,310 --> 03:09:35,945
ONE KEY QUESTION, WHEN DURING

3627
03:09:35,945 --> 03:09:38,915
THE IMMUNE RESPONSE ARE THESE

3628
03:09:38,915 --> 03:09:42,285
ANTIBODIES BEING ELICITED?

3629
03:09:42,285 --> 03:09:43,219
WE PERFORM LONGITUDINAL NEGATIVE

3630
03:09:43,219 --> 03:09:46,422
STAIN TO ANSWER, THE SLIDE IS

3631
03:09:46,422 --> 03:09:47,523
BUSY.

3632
03:09:47,523 --> 03:09:49,192
FIRST WITH THE RABBIT

3633
03:09:49,192 --> 03:09:52,895
IMMUNIZATION, SO THE FIRST

3634
03:09:52,895 --> 03:09:54,030
IMMUNIZATION IN GREEN, THERE'S

3635
03:09:54,030 --> 03:09:55,999
LITTLE TO NO ANTIBODY RESPONSES

3636
03:09:55,999 --> 03:09:57,734
WHICH IS EXPECTED.

3637
03:09:57,734 --> 03:09:59,569
BY THE SECOND IMMUNIZATION WHICH

3638
03:09:59,569 --> 03:10:03,606
IS IN BLUE, WE SEE ANTIBODIES TO

3639
03:10:03,606 --> 03:10:04,707
IMMUNODOMINANT EPITOPES ARISE,

3640
03:10:04,707 --> 03:10:06,609
SO TYPICALLY THE BASE ANTIBODY

3641
03:10:06,609 --> 03:10:10,113
WHICH IS PURPLE HERE AS WELL AS

3642
03:10:10,113 --> 03:10:11,514
SOME GLYCANS, GLYCAN HOLES, AND

3643
03:10:11,514 --> 03:10:14,784
THEN BY THE THIRD IMMUNIZATION

3644
03:10:14,784 --> 03:10:16,753
WE SEE WIDE VARIETY OF EPITOPES

3645
03:10:16,753 --> 03:10:18,955
ELICITED AS WELL AS ANTI-IMMUNE

3646
03:10:18,955 --> 03:10:22,992
COMPLEX ANTIBODIES. 

3647
03:10:22,992 --> 03:10:24,661
FOR THE MACAQUE IMMUNIZATION,

3648
03:10:24,661 --> 03:10:28,464
THIS TREND IS BORNE OUT BUT TOOK

3649
03:10:28,464 --> 03:10:28,731
LONGER.

3650
03:10:28,731 --> 03:10:30,333
THIRD IMMUNIZATION V 5 AND

3651
03:10:30,333 --> 03:10:32,535
INTERFACE FAB APPEAR, BY FOURTH

3652
03:10:32,535 --> 03:10:36,572
YOU SEE THAT CLASS IV

3653
03:10:36,572 --> 03:10:40,543
ANTI-IMMUNE COMPLEX ANTIBODY

3654
03:10:40,543 --> 03:10:40,843
BRIDGING.

3655
03:10:40,843 --> 03:10:41,911
UNFORTUNATELY THIS IS POLYCLONAL

3656
03:10:41,911 --> 03:10:43,780
SO IT'S HARD TO STUDY THESE

3657
03:10:43,780 --> 03:10:46,382
ANTIBODIES IN DETAIL USING OTHER

3658
03:10:46,382 --> 03:10:46,783
ASSAYS.

3659
03:10:46,783 --> 03:10:48,818
BUT PEOPLE IN THE WARD LAB,

3660
03:10:48,818 --> 03:10:50,553
MYSELF INCLUDED, HAVE BEEN

3661
03:10:50,553 --> 03:10:54,524
WORKING ON DIFFERENT STRATEGIES

3662
03:10:54,524 --> 03:10:56,459
TO DETERMINE SEQUENCE OF

3663
03:10:56,459 --> 03:10:59,762
POLYCLONAL ANTIBODIES ONE USING

3664
03:10:59,762 --> 03:11:00,963
SOFTWARE MODELANGELO.

3665
03:11:00,963 --> 03:11:07,236
WE CAN TAKE OUR POLYCLONAL MAP

3666
03:11:07,236 --> 03:11:11,240
AND B CELL RECEPTOR REPERTOIRE,

3667
03:11:11,240 --> 03:11:12,108
AND COMBINE WITH MODELANGELO,

3668
03:11:12,108 --> 03:11:13,743
WILL SPIT OUT A SEQUENCE BASED

3669
03:11:13,743 --> 03:11:18,047
ON OUR FEATURES, AND USE THAT

3670
03:11:18,047 --> 03:11:27,223
INFORMATION TO SEARCH OUR

3671
03:11:27,223 --> 03:11:27,924
REPERTOIRE.

3672
03:11:27,924 --> 03:11:34,330
I WAS ABLE TO FIND DECENT ENOUGH

3673
03:11:34,330 --> 03:11:34,897
MATCHES.

3674
03:11:34,897 --> 03:11:36,299
SO I JUST ORDERED THE

3675
03:11:36,299 --> 03:11:37,533
CONSTRUCTS, WE'LL SEE IF THEY

3676
03:11:37,533 --> 03:11:41,471
EXPRESS AND IF THEY BIND

3677
03:11:41,471 --> 03:11:42,004
APPROPRIATELY.

3678
03:11:42,004 --> 03:11:44,974
SO, I GUESS THE BIG QUESTION IS

3679
03:11:44,974 --> 03:11:47,043
HOW ARE THESE ANTI-IMMUNE

3680
03:11:47,043 --> 03:11:47,677
COMPLEX ANTIBODIES ELICITED?

3681
03:11:47,677 --> 03:11:50,213
AND SO I'LL WALK YOU THROUGH OUR

3682
03:11:50,213 --> 03:11:51,013
CURRENT HYPOTHESIS.

3683
03:11:51,013 --> 03:11:53,616
WE BELIEVE THAT AFTER THE FIRST

3684
03:11:53,616 --> 03:11:55,685
IMMUNIZATION ACTIVATED B CELLS

3685
03:11:55,685 --> 03:11:58,421
ENTER THE GERMINAL CENTER,

3686
03:11:58,421 --> 03:12:00,590
UNDERGO SOMATIC HYPERMUTATION

3687
03:12:00,590 --> 03:12:01,023
AND SELECTION.

3688
03:12:01,023 --> 03:12:08,731
THESE BECOME MEMORY B CELLS PLAS

3689
03:12:08,731 --> 03:12:10,433
MOW BLASTS, TARGETING EPITOPES,

3690
03:12:10,433 --> 03:12:17,073
AND AFTER THE SECONDS

3691
03:12:17,073 --> 03:12:20,443
IMMUNIZATION AID IN FORMATION OF

3692
03:12:20,443 --> 03:12:23,379
IMMUNE COMPLEXES, AND THERE'S

3693
03:12:23,379 --> 03:12:25,415
LITTLE SURFACE AREA LEFT DUE TO

3694
03:12:25,415 --> 03:12:28,017
THIS GLYCOSYLATION AND THESE

3695
03:12:28,017 --> 03:12:34,891
PRIMARY ANTIBODIES, THAT'S

3696
03:12:34,891 --> 03:12:37,059
CALLED EPITOPE MASKING, IN THE

3697
03:12:37,059 --> 03:12:39,595
GERMINAL CENTER THEY ARE

3698
03:12:39,595 --> 03:12:42,064
PRESENTED TO FOLLICULAR

3699
03:12:42,064 --> 03:12:43,666
DENDRITIC CELLS, UNDERGO SOMATIC

3700
03:12:43,666 --> 03:12:48,805
HYPERMUTATION TO BECOME PLAS MOW

3701
03:12:48,805 --> 03:12:55,912
BLASTS AND MEMORY CELLS, CALLED

3702
03:12:55,912 --> 03:12:59,982
AND FORM ANTI-IMMUNE COMPLEXES.

3703
03:12:59,982 --> 03:13:05,555
IMAGING COMPLEXES USING EMPEM

3704
03:13:05,555 --> 03:13:11,260
ALLOWED YOU HAD TO CHARACTERIZE

3705
03:13:11,260 --> 03:13:12,762
THEM, IN FOUR CLASSESES, ARISE

3706
03:13:12,762 --> 03:13:19,335
AFTER THE THIRD AND FOURTH

3707
03:13:19,335 --> 03:13:20,570
IMMUNIZATION, MODELANGEL CAN

3708
03:13:20,570 --> 03:13:23,406
HELP US DETERMINE SEQUENCES OF

3709
03:13:23,406 --> 03:13:26,442
POLYCLONAL ANTIBODIES AND MOST

3710
03:13:26,442 --> 03:13:27,410
IMPORTANTLY THESE ANTIBODIES

3711
03:13:27,410 --> 03:13:28,311
PROBABLY HAVE IMPORTANT

3712
03:13:28,311 --> 03:13:29,412
IMPLICATIONS FOR CURRENT

3713
03:13:29,412 --> 03:13:32,348
APPROACH TO HIV VACCINE DESIGN.

3714
03:13:32,348 --> 03:13:34,984
AND WITH THAT I WANT TO THANK

3715
03:13:34,984 --> 03:13:37,253
THE PEOPLE IN MY LAB WHO

3716
03:13:37,253 --> 03:13:42,592
SUPPORTED THIS WORK, SHOUT OUT

3717
03:13:42,592 --> 03:13:44,293
ALEX, MY POSTDOCTORAL MENTOR,

3718
03:13:44,293 --> 03:13:46,195
NOW RUNS HIS OWN LAB, AIDED IN

3719
03:13:46,195 --> 03:13:48,865
THE COLLECTION OF A LOT OF THIS

3720
03:13:48,865 --> 03:13:49,098
DATA.

3721
03:13:49,098 --> 03:13:49,765
SO THANK YOU.

3722
03:13:49,765 --> 03:13:52,235
[APPLAUSE]

3723
03:13:52,235 --> 03:13:58,040
 

3724
03:13:58,040 --> 03:14:04,213
 

3725
03:14:04,213 --> 03:14:04,780
>> VERY INTERESTING STORY.

3726
03:14:04,780 --> 03:14:09,051
DO YOU KNOW ANYTHING ABOUT THE

3727
03:14:09,051 --> 03:14:11,220
FUNCTION OF THESE ANTI-IMMUNE

3728
03:14:11,220 --> 03:14:13,256
COMPLEX ANTIBODIES, AND

3729
03:14:13,256 --> 03:14:14,390
ESPECIALLY DO THEY INFLUENCE

3730
03:14:14,390 --> 03:14:14,757
NEUTRALIZATION?

3731
03:14:14,757 --> 03:14:15,324
>> GREAT QUESTION.

3732
03:14:15,324 --> 03:14:18,694
SO NONE OF THESE ANIMALS SHOWED

3733
03:14:18,694 --> 03:14:19,529
ANY REMARKABLE NEUTRALIZATION,

3734
03:14:19,529 --> 03:14:21,497
THEY WERE VERY STANDARD FOR

3735
03:14:21,497 --> 03:14:23,699
THEIR COHORTS SO I CAN'T SAY

3736
03:14:23,699 --> 03:14:25,334
THEY WERE BETTER NEUTRALIZERS OR

3737
03:14:25,334 --> 03:14:26,002
WORSE.

3738
03:14:26,002 --> 03:14:27,303
UNFORTUNATELY WE DON'T KNOW A

3739
03:14:27,303 --> 03:14:30,039
LOT ABOUT THEIR FUNCTION BECAUSE

3740
03:14:30,039 --> 03:14:30,706
WE ONLY HAVE POLYCLONAL

3741
03:14:30,706 --> 03:14:34,377
INFORMATION AT THE MOMENT BUT

3742
03:14:34,377 --> 03:14:35,645
WE'RE EXCITED TO HOPEFULLY

3743
03:14:35,645 --> 03:14:40,883
PRODUCE AND START LOOKING AT FC

3744
03:14:40,883 --> 03:14:43,452
RECEPTOR FUNCTION.

3745
03:14:43,452 --> 03:14:48,858
>> ANY OTHER QUESTIONS?

3746
03:14:48,858 --> 03:14:51,260
 

3747
03:14:51,260 --> 03:14:52,328
IF NOT LET'S GIVE SHARIDAN A

3748
03:14:52,328 --> 03:14:53,863
LAST ROUND OF APPLAUSE.

3749
03:14:53,863 --> 03:14:55,531
THANK YOU.

3750
03:14:55,531 --> 03:14:57,934
[APPLAUSE]

3751
03:14:57,934 --> 03:15:01,103
>> NEXT WILL BE
ZHUANROMROM

3752
03:15:01,103 --> 03:15:09,345
YALE UNIVERSITY. 

3753
03:15:09,345 --> 03:15:15,818
 

3754
03:15:15,818 --> 03:15:25,861
 

3755
03:15:36,739 --> 03:15:40,443
>> I WANT TO THANK THE

3756
03:15:40,443 --> 03:15:42,945
ORGANIZERS FOR THE OPPORTUNITY

3757
03:15:42,945 --> 03:15:47,350
TO HEAR OUR WORK.

3758
03:15:47,350 --> 03:15:48,284
I'M ASSOCIATED WITH SCIENTISTS

3759
03:15:48,284 --> 03:15:50,486
IN THE LAB, I'M GOING TO TALK

3760
03:15:50,486 --> 03:16:00,896
ABOUT THE DYNAMIC OF THE

3761
03:16:04,767 --> 03:16:05,935
MEMBRANE-PROXIMAL EXTERNAL

3762
03:16:05,935 --> 03:16:06,168
REGION.

3763
03:16:06,168 --> 03:16:08,004
PRIMARY GOAL TO INTRODUCE

3764
03:16:08,004 --> 03:16:11,774
BROADLY NEUTRALIZING ANTIBODIES

3765
03:16:11,774 --> 03:16:15,378
THAT PROTECT AGAINST HIV

3766
03:16:15,378 --> 03:16:18,414
STRAINS. 

3767
03:16:18,414 --> 03:16:21,150
THIS BROADLY NEUTRALIZING

3768
03:16:21,150 --> 03:16:24,320
ANTIBODY, ENVELOPE GLYCOPROTEIN,

3769
03:16:24,320 --> 03:16:29,225
INCLUDING THE MEMBRANE

3770
03:16:29,225 --> 03:16:32,662
PROXIMAL-EXTERNAL REGION, MPER.

3771
03:16:32,662 --> 03:16:35,131
AMONG THOSE ANTIBODIES THE

3772
03:16:35,131 --> 03:16:45,675
ANTI-MPER ANTIBODY STAND OUT FOR

3773
03:16:52,281 --> 03:16:52,448
POTENCY.

3774
03:16:52,448 --> 03:16:54,350
TWO REASON STUDY SHOW MPER

3775
03:16:54,350 --> 03:16:58,454
EPITOPE CAN BE USED IN VACCINE

3776
03:16:58,454 --> 03:16:58,888
DEVELOPMENT.

3777
03:16:58,888 --> 03:17:01,724
AND THOSE TWO STUDIES

3778
03:17:01,724 --> 03:17:05,561
SUCCESSFULLY INDUCED BROADLY

3779
03:17:05,561 --> 03:17:08,497
NEUTRALIZING ANTIBODY.

3780
03:17:08,497 --> 03:17:13,436
IN PREVIOUS STUDIES THE ANTIBODY

3781
03:17:13,436 --> 03:17:15,838
FAB BINDS TO ENVELOPE HAS BEEN

3782
03:17:15,838 --> 03:17:21,177
VISUALIZED USING CryoEM SINGLE

3783
03:17:21,177 --> 03:17:24,213
PARTICLE ANALYSIS. 

3784
03:17:24,213 --> 03:17:25,581
TWO STUDIES OF THE DIFFERENT

3785
03:17:25,581 --> 03:17:29,118
STATES OF ONE, TWO, AND THREE

3786
03:17:29,118 --> 03:17:33,823
MPER ANTIBODY 28 FAB AND 4010

3787
03:17:33,823 --> 03:17:40,262
FAB, SHOW DIFFERENT -- SHOWING

3788
03:17:40,262 --> 03:17:43,199
DIFFERENT DISTANCE OF ENVELOPE

3789
03:17:43,199 --> 03:17:45,167
FROM THE MEMBRANE.

3790
03:17:45,167 --> 03:17:49,438
THEY FOUND ONE FAB COMBINES TO

3791
03:17:49,438 --> 03:17:54,744
THE ENVELOPE AND THREE FABS

3792
03:17:54,744 --> 03:18:01,684
COMBINE TO ENVELOPE FROM THE

3793
03:18:01,684 --> 03:18:02,118
MEMBRANE.

3794
03:18:02,118 --> 03:18:06,689
THE STUDY ALSO SHOWS THAT

3795
03:18:06,689 --> 03:18:08,491
DIFFERENT MEMBRANES SOMETIMES

3796
03:18:08,491 --> 03:18:12,428
HAVE DIFFERENT CUMULATIVE WORK,

3797
03:18:12,428 --> 03:18:15,798
ROTATIONAL POSITION AND

3798
03:18:15,798 --> 03:18:19,268
COMPOSITIONAL PROPERTY,

3799
03:18:19,268 --> 03:18:25,841
ADDITIONALLY THE MPER ANTIBODY

3800
03:18:25,841 --> 03:18:28,110
SHARE -- THEY ALIGN MPER EPITOPE

3801
03:18:28,110 --> 03:18:32,915
FROM CRITICAL INFRASTRUCTURE

3802
03:18:32,915 --> 03:18:33,048
,

3803
03:18:33,048 --> 03:18:37,753
BINDS TO ANTIBODY IN A SIMILAR

3804
03:18:37,753 --> 03:18:38,387
WAY.

3805
03:18:38,387 --> 03:18:39,054
THOSE STUDIES USE PURIFIED

3806
03:18:39,054 --> 03:18:39,388
ENVELOPE.

3807
03:18:39,388 --> 03:18:45,528
WE WANT TO LOOK AT THE ENVELOPE

3808
03:18:45,528 --> 03:18:47,496
FROM MULTIPLE VIRUS AND STUDY

3809
03:18:47,496 --> 03:18:48,464
PROPERTY.

3810
03:18:48,464 --> 03:18:54,170
IN OUR STUDY WE USE THE MPER

3811
03:18:54,170 --> 03:18:59,842
ANTIBODY FAB, TH 511.

3812
03:18:59,842 --> 03:19:06,315
WE FOUND IN ABSENCE OF 34 YOU

3813
03:19:06,315 --> 03:19:12,755
CAN ALREADY SEE THE DH 511

3814
03:19:12,755 --> 03:19:15,291
BINDING FROM THE TOMOGRAM.

3815
03:19:15,291 --> 03:19:17,593
THEN WE DID IMAGING ON THE

3816
03:19:17,593 --> 03:19:20,663
CLASSIFICATION, WE FOUND HALF OF

3817
03:19:20,663 --> 03:19:26,468
THE ENVELOPES DON'T HAVE DH 511,

3818
03:19:26,468 --> 03:19:32,041
AND ABOUT A HALF HAS THREE DL511

3819
03:19:32,041 --> 03:19:33,676
BIND SITE.

3820
03:19:33,676 --> 03:19:43,452
WE DID NOT OBSERVE ANY  ONE OR

3821
03:19:43,452 --> 03:19:44,386
TWO BINDING.

3822
03:19:44,386 --> 03:19:48,023
SO THEN WE THOUGHT AVERAGE

3823
03:19:48,023 --> 03:19:50,960
STRUCTURE, TO AVERAGE MAP, WE

3824
03:19:50,960 --> 03:19:52,294
FOUND THE MEMBRANE

3825
03:19:52,294 --> 03:19:53,929
REORGANIZATION IN THE CENTRAL

3826
03:19:53,929 --> 03:19:57,766
SLICE, WE ALREADY KNOW THAT THE

3827
03:19:57,766 --> 03:20:00,169
MPER REGION PLAY A ROLE IN THE

3828
03:20:00,169 --> 03:20:05,074
MEMBRANE FUSION AND SAW HOW THIS

3829
03:20:05,074 --> 03:20:08,677
AMPLIFIED FAB BINDING CAUSE THE

3830
03:20:08,677 --> 03:20:10,746
THIS MEMBRANE.

3831
03:20:10,746 --> 03:20:12,681
ALL RIGHT.

3832
03:20:12,681 --> 03:20:21,523
THIS IS THE CRYSTAL STRUCTURE OF

3833
03:20:21,523 --> 03:20:25,294
THE DH511 FAB BINDS TO THE AMPER

3834
03:20:25,294 --> 03:20:31,100
AND PLACE THIS STRUCTURE INTO

3835
03:20:31,100 --> 03:20:36,672
OUR ELECTRON DENSITY, THIS IS

3836
03:20:36,672 --> 03:20:39,541
THE 3D RENDERING.

3837
03:20:39,541 --> 03:20:43,913
WE USE THE CRYSTAL STRUCTURE

3838
03:20:43,913 --> 03:20:45,981
FROM DH511 FAB AND CLOSE THE

3839
03:20:45,981 --> 03:20:47,316
ENVELOPE STRUCTURE.

3840
03:20:47,316 --> 03:20:50,019
YOU CAN SEE THE MEMBRANE

3841
03:20:50,019 --> 03:20:58,027
DISTORTION IS RIGHT IN CENTER,

3842
03:20:58,027 --> 03:20:59,595
UNDERNEATH THE ENVELOPE.

3843
03:20:59,595 --> 03:21:01,897
AND TO FACILITATE THIS BINDING,

3844
03:21:01,897 --> 03:21:03,999
THIS EPITOPE NEED TO BE PLACED

3845
03:21:03,999 --> 03:21:10,272
AT ONE SIDE OF THE ENVELOPE.

3846
03:21:10,272 --> 03:21:14,643
HOWEVER, IN THE LAMAR STRUCTURE,

3847
03:21:14,643 --> 03:21:19,014
THE REGION FORMS SYMMETRIC ABOUT

3848
03:21:19,014 --> 03:21:21,216
THE TRANSMEMBRANE DOMAIN,

3849
03:21:21,216 --> 03:21:25,187
DIFFERENT, THE MPER EPITOPE IN

3850
03:21:25,187 --> 03:21:32,761
THE STRUCTURE IS DIFFERENT FROM

3851
03:21:32,761 --> 03:21:35,564
EPITOPE IN OUR STRUCTURE.

3852
03:21:35,564 --> 03:21:44,039
MPER NEED TO BE ON ONE SIDE OF

3853
03:21:44,039 --> 03:21:46,208
THE ENVELOPE.

3854
03:21:46,208 --> 03:21:51,113
SO WE'RE USING THIS TO STUDY,

3855
03:21:51,113 --> 03:21:54,183
THE REASON WE USE STIMULATION

3856
03:21:54,183 --> 03:21:58,320
ALTHOUGH PART 69 EPITOPE IS

3857
03:21:58,320 --> 03:22:01,256
STABILIZED, THE OTHER PART THAT

3858
03:22:01,256 --> 03:22:03,359
CONNECTING TO THE ENVELOPE IS

3859
03:22:03,359 --> 03:22:07,463
STILL FLEXIBLE SO WHEN I ALIGN

3860
03:22:07,463 --> 03:22:11,800
THE TOP ENVELOPE PART FROM THE

3861
03:22:11,800 --> 03:22:18,007
GROUND YOU CAN SEE THE DH511 FAB

3862
03:22:18,007 --> 03:22:21,076
IS SHIFTING, WHEN ALIGN DH 511

3863
03:22:21,076 --> 03:22:23,846
FAB AT THE BOTTOM OF THE

3864
03:22:23,846 --> 03:22:28,650
SUBTOMOGRAM YOU CAN SEE THE

3865
03:22:28,650 --> 03:22:36,325
EPITOPE SHIFTS AND ROTATES.

3866
03:22:36,325 --> 03:22:36,959
 

3867
03:22:36,959 --> 03:22:42,698
NEXT I ANALYZE THE FAB BIND

3868
03:22:42,698 --> 03:22:48,604
ENVELOPE AND FOUND THE FREQUENCY

3869
03:22:48,604 --> 03:22:49,905
PEAK AT ABOUT 30-DEGREE.

3870
03:22:49,905 --> 03:22:54,176
COME DOWN TO THE UNBOUND

3871
03:22:54,176 --> 03:23:01,617
ENVELOPE, FREQUENCY PEAK AT 12

3872
03:23:01,617 --> 03:23:02,017
DEGREE.

3873
03:23:02,017 --> 03:23:08,290
NOW WE HAVE THE QUESTION ARE THE

3874
03:23:08,290 --> 03:23:11,560
ENVELOPE ALREADY INHIBITED?

3875
03:23:11,560 --> 03:23:13,529
TO ANSWER THIS QUESTION WE

3876
03:23:13,529 --> 03:23:17,366
PREPARED ANOTHER SAMPLE IN WHICH

3877
03:23:17,366 --> 03:23:23,605
WE ADDED DH511 FIRST, CD4 AND

3878
03:23:23,605 --> 03:23:24,139
17B.

3879
03:23:24,139 --> 03:23:28,644
WE FOUND ONE-THIRD -- WE FOUND A

3880
03:23:28,644 --> 03:23:34,783
HALF -- WE FOUND A HALF OF THE

3881
03:23:34,783 --> 03:23:38,554
ENVELOPE HAS -- ALL OF THE 17B

3882
03:23:38,554 --> 03:23:42,791
BOUND AND HALF OF THE DH511 FAB

3883
03:23:42,791 --> 03:23:44,827
BINDS FROM ONE SIDE.

3884
03:23:44,827 --> 03:23:55,370
AND ABOUT A THIRD HAS THE DH 511

3885
03:23:57,406 --> 03:24:00,175
FAB BINDS SYMMETRICALLY.

3886
03:24:00,175 --> 03:24:04,246
SO THE CD4 AND 17B TO STILL BIND

3887
03:24:04,246 --> 03:24:07,616
TO THE ENVELOPE SUGGESTING THAT

3888
03:24:07,616 --> 03:24:10,219
THE ENVELOPE STILL UNDERGOES CD4

3889
03:24:10,219 --> 03:24:11,487
DEPENDENT ACTIVATION.

3890
03:24:11,487 --> 03:24:13,589
ALSO FROM THE NUMBERS YOU CAN

3891
03:24:13,589 --> 03:24:18,794
SEE THE CDs 4 BINDING INCREASE

3892
03:24:18,794 --> 03:24:26,468
DH511 BINDING, ALREADY REPORTED

3893
03:24:26,468 --> 03:24:28,504
IN LITERATURE.

3894
03:24:28,504 --> 03:24:31,440
HOWEVER THIS ENVELOPE STILL

3895
03:24:31,440 --> 03:24:36,612
RETAIN FLEXIBILITY, SO FROM THE

3896
03:24:36,612 --> 03:24:43,952
ANALYSIS WE CAN SEE THE CD4 AND

3897
03:24:43,952 --> 03:24:46,054
THE 17B BINDS ENVELOPE WITHOUT

3898
03:24:46,054 --> 03:24:53,829
FAB LARGER RAIN OF TRT AND THE

3899
03:24:53,829 --> 03:24:57,900
DH511 BINDING OF FAB ENVELOPE,

3900
03:24:57,900 --> 03:25:07,309
START BINDING ENVELOPE,

3901
03:25:07,309 --> 03:25:08,911
CONJOINED SYMMETRICALLY IN

3902
03:25:08,911 --> 03:25:09,378
ENVELOPE.

3903
03:25:09,378 --> 03:25:13,715
IN SUMMARY, WE HAVE SHOWN THAT

3904
03:25:13,715 --> 03:25:20,622
THE DH 511 COULD BIND TO OPEN

3905
03:25:20,622 --> 03:25:27,596
AND CLOSED ENVELOPE.

3906
03:25:27,596 --> 03:25:31,333
NEED TO BE SHIFTED AT ONE SIDE

3907
03:25:31,333 --> 03:25:34,736
OF THE ENVELOPE, SO INDICATING

3908
03:25:34,736 --> 03:25:37,472
IF EXCEPTIONAL CONFIRMATION OF

3909
03:25:37,472 --> 03:25:48,016
FLEXIBILITY OF THE MPER REGION.

3910
03:25:55,524 --> 03:25:59,161
THE ENVELOPE CAN UNDERGO

3911
03:25:59,161 --> 03:25:59,695
CD4-DEPENDENT ACTIVATION.

3912
03:25:59,695 --> 03:26:02,497
HOWEVER, LOOK AT ALL THOSE

3913
03:26:02,497 --> 03:26:04,233
STRUCTURES WE STILL HAVE MANY

3914
03:26:04,233 --> 03:26:08,270
QUESTIONS.

3915
03:26:08,270 --> 03:26:11,907
SO FIRST THAT'S THE MPER

3916
03:26:11,907 --> 03:26:12,874
ANTIBODY INDUCED ENVELOPE

3917
03:26:12,874 --> 03:26:19,448
CHANGING, JUST SIMPLY BINDS TO

3918
03:26:19,448 --> 03:26:27,956
TRT ENVELOPE.

3919
03:26:27,956 --> 03:26:32,894
ADDITIONAL DH511 CD4, CAN EITHER

3920
03:26:32,894 --> 03:26:39,201
BE FRONT OF TRT ENVELOPE.

3921
03:26:39,201 --> 03:26:40,569
GIVE THIS STRUCTURE.

3922
03:26:40,569 --> 03:26:46,908
AND THE LAST BUT NOT THE LEAST

3923
03:26:46,908 --> 03:26:56,618
CAN THIS TWO STRUCTURE

3924
03:26:56,618 --> 03:26:57,252
TRANSITIONAL CONFIRMATIONAL

3925
03:26:57,252 --> 03:26:57,619
EQUILIBRIUM.

3926
03:26:57,619 --> 03:27:01,857
I WANT TO THANK ALL THE LAB

3927
03:27:01,857 --> 03:27:03,525
MEMBERS AND OUR COLLABORATORS.

3928
03:27:03,525 --> 03:27:04,092
THANK YOU.

3929
03:27:04,092 --> 03:27:07,396
[APPLAUSE]

3930
03:27:07,396 --> 03:27:12,701
 

3931
03:27:12,701 --> 03:27:18,173
 

3932
03:27:18,173 --> 03:27:18,974
>> GREAT WORK.

3933
03:27:18,974 --> 03:27:23,979
I HAVE A QUESTION ABOUT THE

3934
03:27:23,979 --> 03:27:24,613
DIFFERENCE BETWEEN THE STRUCTURE

3935
03:27:24,613 --> 03:27:25,414
AND STRUCTURE WITH THE

3936
03:27:25,414 --> 03:27:27,449
ANTIBODIES, DO YOU THINK THE

3937
03:27:27,449 --> 03:27:29,117
ANTIBODY BINDING IS CHANGING

3938
03:27:29,117 --> 03:27:30,752
THAT MPER OR IS JUST THAT SINCE

3939
03:27:30,752 --> 03:27:36,558
YOUR SYSTEM IS A LITTLE BIT MORE

3940
03:27:36,558 --> 03:27:38,894
NATIVE THAT IS ACTUALLY THE

3941
03:27:38,894 --> 03:27:40,162
STRUCTURAL WITH THE MPER, THEN

3942
03:27:40,162 --> 03:27:42,164
MY SECOND YES IF IT'S THE

3943
03:27:42,164 --> 03:27:43,999
ANTIBODIES THAT ARE CAUSING THAT

3944
03:27:43,999 --> 03:27:46,101
CHANGE HOW DOES THAT TRANSLATE

3945
03:27:46,101 --> 03:27:49,037
TO THE TRANSMEMBRANE REGION AND

3946
03:27:49,037 --> 03:27:51,573
THE CYTOPLASMIC TAIL OF THE

3947
03:27:51,573 --> 03:27:54,076
ENVELOPE, BASICALLY CAN MPER

3948
03:27:54,076 --> 03:27:55,410
BINDING CREATE PROPAGATING

3949
03:27:55,410 --> 03:27:57,279
CHANGES THAT CHANGE THE

3950
03:27:57,279 --> 03:27:58,814
CYTOPLASMIC TAIL OF THE ENVELOPE

3951
03:27:58,814 --> 03:28:02,551
AND MAYBE THAT'S WHY MPER

3952
03:28:02,551 --> 03:28:03,518
ANTIBODIES ARE SO GOOD?

3953
03:28:03,518 --> 03:28:05,487
>> THAT'S A VERY GOOD
QUESTION.

3954
03:28:05,487 --> 03:28:09,891
WE DON'T KNOW THAT. 

3955
03:28:09,891 --> 03:28:14,229
>> OKAY, THANK YOU. 

3956
03:28:14,229 --> 03:28:14,830
>> GO AHEAD.

3957
03:28:14,830 --> 03:28:17,199
>> VERY NICE.

3958
03:28:17,199 --> 03:28:20,135
DO YOU SEE ANY NEAREST NEIGHBOR

3959
03:28:20,135 --> 03:28:22,471
EFFECT, LIKE 50% BOUND, 50%

3960
03:28:22,471 --> 03:28:24,272
UNBOUND, ARE BOUND ONES

3961
03:28:24,272 --> 03:28:28,110
CLUSTERED TOGETHER OR IS IT

3962
03:28:28,110 --> 03:28:29,378
PRETTY RANDOM?

3963
03:28:29,378 --> 03:28:31,012
>> FROM THE -- YEAH,
SOMEONE

3964
03:28:31,012 --> 03:28:37,986
ELSE ASK ME THIS QUESTION. 

3965
03:28:37,986 --> 03:28:41,490
FROM THE TOMOGRAM YOU SEE THEY

3966
03:28:41,490 --> 03:28:42,224
ARE CLUSTERING.

3967
03:28:42,224 --> 03:28:43,992
YOU'LL SEE THEM CLUSTERING.

3968
03:28:43,992 --> 03:28:45,193
>> THEY ARE CLUSTERED
TOGETHER?

3969
03:28:45,193 --> 03:28:49,998
>> YEAH, BY THE WAY WE
NEED LIKE

3970
03:28:49,998 --> 03:28:52,834
ANALYZE THE DATA THAT YOU GIVE A

3971
03:28:52,834 --> 03:28:53,135
CONCLUSION.

3972
03:28:53,135 --> 03:28:59,541
>> THANK YOU. 

3973
03:28:59,541 --> 03:29:03,378
 

3974
03:29:03,378 --> 03:29:07,382
>> THANK YOU AGAIN, ZHUAN.

3975
03:29:07,382 --> 03:29:07,649
[APPLAUSE]

3976
03:29:07,649 --> 03:29:09,217
NEXT IS JAMIN WILLOUGHBY,

3977
03:29:09,217 --> 03:29:17,826
UNIVERSITY OF UTAH. 

3978
03:29:17,826 --> 03:29:27,869
 

3979
03:30:00,235 --> 03:30:03,071
>> TECHNICAL DIFFICULTIES.

3980
03:30:03,071 --> 03:30:07,142
 

3981
03:30:07,142 --> 03:30:17,185
 

3982
03:32:12,934 --> 03:32:16,571
>> SWEET, OKAY.

3983
03:32:16,571 --> 03:32:19,274
 

3984
03:32:19,274 --> 03:32:22,844
I'M A SECOND YEAR GRADUATE

3985
03:32:22,844 --> 03:32:25,080
STUDENT IN THE STAR LABS,

3986
03:32:25,080 --> 03:32:25,547
UNIVERSITY OF UTAH.

3987
03:32:25,547 --> 03:32:27,816
I'D LIKE TO THANK THE ORGANIZERS

3988
03:32:27,816 --> 03:32:29,484
FOR INVITING ME TO GIVE A LITTLE

3989
03:32:29,484 --> 03:32:29,684
TALK.

3990
03:32:29,684 --> 03:32:31,419
I'D LIKE TO TELL YOU A BIT OF A

3991
03:32:31,419 --> 03:32:33,722
STORY HOW I STARTED TO MAP

3992
03:32:33,722 --> 03:32:42,263
MUTATIONAL LANDSCAPE OF BROADLY

3993
03:32:42,263 --> 03:32:43,264
NEUTRALIZING ANTIBODIES,

3994
03:32:43,264 --> 03:32:45,867
bNAbS ARISE FROM THE ARMS

3995
03:32:45,867 --> 03:32:48,169
RACE, TRYING TO EVOLVE TO COMBAT

3996
03:32:48,169 --> 03:32:51,706
A POOL OF HIV IN THE HOST, AND

3997
03:32:51,706 --> 03:32:54,609
SO THEREFORE THEY DEVELOP THE

3998
03:32:54,609 --> 03:32:56,645
ABILITY TO NEUTRALIZE

3999
03:32:56,645 --> 03:32:57,946
HETEROLOGOUS HIV ISOLATES FROM

4000
03:32:57,946 --> 03:32:59,447
OTHER INDIVIDUALS THROUGHOUT THE

4001
03:32:59,447 --> 03:33:00,682
WORLD.

4002
03:33:00,682 --> 03:33:04,953
AND THE ANTIBODIES THAT TARGET

4003
03:33:04,953 --> 03:33:06,888
CD4 BINDING SITE ARE BROAD

4004
03:33:06,888 --> 03:33:10,659
BECAUSE IT'S HIGHLY CONSERVED

4005
03:33:10,659 --> 03:33:13,461
VITAL FOR CELL ENTRY AND MIMIC

4006
03:33:13,461 --> 03:33:14,596
CD4 RECEPTOR.

4007
03:33:14,596 --> 03:33:23,038
HERE IN SORT OF A STRUCTURAL

4008
03:33:23,038 --> 03:33:25,140
COMPARISON BETWEEN CD4 AND HEAVY

4009
03:33:25,140 --> 03:33:26,207
CHAIN IMMUNIZING IMMUNOGLOBULIN

4010
03:33:26,207 --> 03:33:28,209
FOLD THAT'S TYPICAL OF THE CD4

4011
03:33:28,209 --> 03:33:28,476
RECEPTOR.

4012
03:33:28,476 --> 03:33:30,045
SO THEREFORE A LOT OF ANTIBODIES

4013
03:33:30,045 --> 03:33:32,347
ARE ABLE TO NEUTRALIZE OVER 90%

4014
03:33:32,347 --> 03:33:34,215
OF GLOBAL HIV STRAINS.

4015
03:33:34,215 --> 03:33:38,153
AND OVER THE YEARS OF MORE SORT

4016
03:33:38,153 --> 03:33:40,021
OF -- AS PEOPLE BEGAN TO ISOLATE

4017
03:33:40,021 --> 03:33:42,524
ANTIBODIES WITH MORE BREADTH AND

4018
03:33:42,524 --> 03:33:45,060
POTENCY BEGAN TO SEE MANY

4019
03:33:45,060 --> 03:33:46,695
SIMILAR bNAbS AROSE IN

4020
03:33:46,695 --> 03:33:48,596
DIFFERENT DONORS, SUGGESTING A

4021
03:33:48,596 --> 03:33:49,297
CONVERGENT EVOLUTIONARY THING

4022
03:33:49,297 --> 03:33:50,732
THAT WE MIGHT BE ABLE TO DIRECT,

4023
03:33:50,732 --> 03:33:53,168
SHOWN HERE WITH A STRUCTURAL

4024
03:33:53,168 --> 03:33:55,036
OVERLAP OF FIVE DIFFERENT CD4

4025
03:33:55,036 --> 03:34:00,842
BINDING SITE bNAbS FROM FIVE

4026
03:34:00,842 --> 03:34:02,110
DONORS TARGETING GP120.

4027
03:34:02,110 --> 03:34:04,946
THE COMMON BINDING MODE THAT

4028
03:34:04,946 --> 03:34:06,147
DICTATES INTERACTION IS

4029
03:34:06,147 --> 03:34:08,016
STRUCTURALLY CONSERVED AMONGST

4030
03:34:08,016 --> 03:34:09,451
THESE ANTIBODIES.

4031
03:34:09,451 --> 03:34:13,288
FURTHERMORE THIS IS ENCODED

4032
03:34:13,288 --> 03:34:14,589
COMMONLY BY THE VH1-2 GENE WHICH

4033
03:34:14,589 --> 03:34:19,494
MOST INDIVIDUALS IN THE WORLD

4034
03:34:19,494 --> 03:34:19,761
HAVE.

4035
03:34:19,761 --> 03:34:22,363
IT HIGHLIGHTS THAT WE MIGHT

4036
03:34:22,363 --> 03:34:26,401
LEVERAGE FOR EFFECTIVE VACCINE

4037
03:34:26,401 --> 03:34:29,671
TO INDUCE CD4 BINDING SITES.

4038
03:34:29,671 --> 03:34:30,872
TRYING TO ELICIT THESE

4039
03:34:30,872 --> 03:34:32,407
ANTIBODIES, THEY HAVE BEEN USING

4040
03:34:32,407 --> 03:34:33,842
SORT OF THE MATURE ANTIBODY

4041
03:34:33,842 --> 03:34:37,011
PROTOTYPES THEY HAVE ISOLATED AS

4042
03:34:37,011 --> 03:34:39,247
BENCH MARKS FOR SUCCESS.

4043
03:34:39,247 --> 03:34:44,385
ALL THESE IMMUNOGENS HAVE SO FAR

4044
03:34:44,385 --> 03:34:48,556
FAILED TO ELICIT IN A BROAD AND

4045
03:34:48,556 --> 03:34:49,190
PROTECTIVE MANNER HIGHLIGHTING

4046
03:34:49,190 --> 03:34:50,391
CRE FEATURES THAT WE HAVE TO

4047
03:34:50,391 --> 03:34:52,460
TAKE INTO ACCOUNT AND HAVE BEEN

4048
03:34:52,460 --> 03:34:55,964
YET TO BE OVERCOME DEPENDABLY.

4049
03:34:55,964 --> 03:34:57,398
THERE WE GO.

4050
03:34:57,398 --> 03:35:00,969
SO, FIRST AND FOREMOST THESE

4051
03:35:00,969 --> 03:35:08,309
ANTIBODIES ARE EXTENSIVELY

4052
03:35:08,309 --> 03:35:10,945
MUTATED FROM NAIVE PRECURSORS,

4053
03:35:10,945 --> 03:35:12,147
HARDER TO CAPITULATE IN A

4054
03:35:12,147 --> 03:35:16,951
SHORTER TIME SPAN.

4055
03:35:16,951 --> 03:35:19,354
THERE'S PRESENCE OF ISOLATION AT

4056
03:35:19,354 --> 03:35:26,828
276, THIS GLYCOSYLATION IS

4057
03:35:26,828 --> 03:35:28,429
PRESENT IN 95%.

4058
03:35:28,429 --> 03:35:30,098
DURING THE EVOLUTION OF A LOT OF

4059
03:35:30,098 --> 03:35:37,405
THESE ANTIBODIES THEY KIND OF

4060
03:35:37,405 --> 03:35:41,142
TRIED TO ALEVE BY TRUNCATING

4061
03:35:41,142 --> 03:35:43,878
LOOPS, YET TO BE DEPENDABLY

4062
03:35:43,878 --> 03:35:47,248
ELICITED IN MOST VACCINE

4063
03:35:47,248 --> 03:35:47,515
RESPONSES.

4064
03:35:47,515 --> 03:35:48,917
SO THIS HIGHLIGHTS TWO THINGS

4065
03:35:48,917 --> 03:35:56,958
THAT WE NEED TO KNOW HOW ALL THE

4066
03:35:56,958 --> 03:36:03,898
POSSIBLE WAY THE 2376 GLYCAN.

4067
03:36:03,898 --> 03:36:05,300
A MORE HOLISTIC ANALYSIS THAT

4068
03:36:05,300 --> 03:36:06,267
THEY CAN PICK UP ALONG THE WAY

4069
03:36:06,267 --> 03:36:10,071
WOULD BE ABLE TO INFORM HOW NEXT

4070
03:36:10,071 --> 03:36:11,039
GEN IMMUNOGENS COULD ELICIT

4071
03:36:11,039 --> 03:36:11,739
PATHWAYS WE WANT.

4072
03:36:11,739 --> 03:36:13,708
WE WOULD LIKE TO TRY AND ELICIT

4073
03:36:13,708 --> 03:36:15,877
BROAD AND POTENT ANTIBODIES WITH

4074
03:36:15,877 --> 03:36:19,047
MINIMAL MUTATIONS NECESSARY TO

4075
03:36:19,047 --> 03:36:21,249
BE BROAD, POTENT, PROTECTIVE.

4076
03:36:21,249 --> 03:36:24,285
THIS LEADS ME TO THE IOMA CLASS,

4077
03:36:24,285 --> 03:36:29,557
AND SO THIS CLASS IS

4078
03:36:29,557 --> 03:36:31,392
CHARACTERIZED IN 2016, AND

4079
03:36:31,392 --> 03:36:36,431
CONTRARY TO MOST COMMON CD4

4080
03:36:36,431 --> 03:36:37,832
BINDING SITE bNAbS, THE

4081
03:36:37,832 --> 03:36:40,368
CLASS OF ANTIBODIES THAT HAS

4082
03:36:40,368 --> 03:36:42,470
BEEN GIVEN MUCH EFFORT INTO

4083
03:36:42,470 --> 03:36:46,875
VACCINE ELICITATION, THEY HAVE

4084
03:36:46,875 --> 03:36:51,546
ACQUIRED THE IOMA -- THE CLASS

4085
03:36:51,546 --> 03:36:56,584
ACQUIRED MORE, AND HAS A NORMAL

4086
03:36:56,584 --> 03:36:58,586
LENGTH CDR L1 THAT CAN

4087
03:36:58,586 --> 03:37:00,655
ACCOMMODATE THE GLYCAN THROUGH A

4088
03:37:00,655 --> 03:37:02,523
DIFFERENT MECHANISM, SHOWN HERE

4089
03:37:02,523 --> 03:37:05,960
IN THIS FIGURE WHERE IOMA HAS A

4090
03:37:05,960 --> 03:37:08,196
10% SEQUENCE DIVERSITY FROM

4091
03:37:08,196 --> 03:37:18,606
NAIVE ANTIBODY PRECURSOR

4092
03:37:21,743 --> 03:37:22,443
COMPARES TO 25%.

4093
03:37:22,443 --> 03:37:28,583
WE'VE ONLY BEEN ABLE TO

4094
03:37:28,583 --> 03:37:31,219
NEUTRALIZE 25%, IT HAS LESS

4095
03:37:31,219 --> 03:37:33,821
NECESSARY MUTATIONS ALONG THE

4096
03:37:33,821 --> 03:37:34,055
WAY.

4097
03:37:34,055 --> 03:37:39,394
AND THE BJORKMAN GROUP HAS SHOWN

4098
03:37:39,394 --> 03:37:44,232
THIS TO SUCCESS A COUPLE YEARS

4099
03:37:44,232 --> 03:37:48,503
AGO DEVELOPING TWO IGT 1 AND 2

4100
03:37:48,503 --> 03:37:50,505
IMMUNOGENS, IN THE ANTIBODIES

4101
03:37:50,505 --> 03:37:51,906
THEY ELICITED IN THEIR STUDY

4102
03:37:51,906 --> 03:37:54,475
THEY DID HAVE SOME POTENT

4103
03:37:54,475 --> 03:37:55,510
NEUTRALIZING ACTIVITY.

4104
03:37:55,510 --> 03:37:57,178
AND SO I'VE BEEN COLLABORATING

4105
03:37:57,178 --> 03:37:59,147
WITH THE BJORKMAN GROUP TO

4106
03:37:59,147 --> 03:37:59,781
COMPLEMENT RESULTS AND

4107
03:37:59,781 --> 03:38:01,883
VACCINATION STUDIES USING MORE

4108
03:38:01,883 --> 03:38:03,751
UNBIASED MUTATIONAL SCANS TO

4109
03:38:03,751 --> 03:38:05,720
HOLISTICALLY DETERMINE HOW ALL

4110
03:38:05,720 --> 03:38:08,089
THE POSSIBLE MUTATIONS IN THIS

4111
03:38:08,089 --> 03:38:11,159
IOMA ANTIBODY CAN AFFECT THE

4112
03:38:11,159 --> 03:38:13,695
POTENCY OF RECOGNITION, TRY AND

4113
03:38:13,695 --> 03:38:17,966
ENGINEER IOMA TO BE MORE BROAD

4114
03:38:17,966 --> 03:38:20,068
AND POTENT, IT CAN ALSO AID THE

4115
03:38:20,068 --> 03:38:24,072
NEXT GENERATION OF IMMUNOGENS TO

4116
03:38:24,072 --> 03:38:25,273
TARGET IOMA CLASS PAINTING A

4117
03:38:25,273 --> 03:38:26,841
BETTER TARGET OF WHAT WE MIGHT

4118
03:38:26,841 --> 03:38:27,775
WANT TO SHOOT FOR.

4119
03:38:27,775 --> 03:38:29,744
THAT LEADS ME TO WHAT I WANT TO

4120
03:38:29,744 --> 03:38:31,279
PROPOSE, WHERE I'D LIKE TO USE

4121
03:38:31,279 --> 03:38:32,580
DEEP MUTATIONAL SCANNING TO

4122
03:38:32,580 --> 03:38:35,316
DETERMINE HOW THE POSSIBLE

4123
03:38:35,316 --> 03:38:36,985
MUTATIONS IOMA CAN ACCRUE, WHAT

4124
03:38:36,985 --> 03:38:41,122
THEIR EFFECT WOULD BE ON BINDING

4125
03:38:41,122 --> 03:38:41,856
AFFINITY.

4126
03:38:41,856 --> 03:38:44,325
DEEP MUTATIONAL SCANNING ALLOWS

4127
03:38:44,325 --> 03:38:46,728
US TO SAMPLE MUTATIONS AND

4128
03:38:46,728 --> 03:38:48,730
DELETIONS IN A PROTEIN OF

4129
03:38:48,730 --> 03:38:50,698
INTEREST AND BE APPLIED TO THE

4130
03:38:50,698 --> 03:38:52,300
HEAVY AND LIGHT CHAINS OF

4131
03:38:52,300 --> 03:38:52,600
ANTIBODIES.

4132
03:38:52,600 --> 03:38:57,238
TO DO THIS, I CAN TRANSFORM A

4133
03:38:57,238 --> 03:38:59,807
SINGLE MUTANT PROTEIN LIBRARY OF

4134
03:38:59,807 --> 03:39:02,477
THE IOMA INTO YEAST WHERE THEY

4135
03:39:02,477 --> 03:39:04,479
CAN DISPLAY THAT PROTEIN ON

4136
03:39:04,479 --> 03:39:06,314
THEIR CELL SURFACE, IN LIKE A

4137
03:39:06,314 --> 03:39:08,182
SINGLE OPEN READING FRAME BY

4138
03:39:08,182 --> 03:39:09,951
EXPRESSING ANTIBODIES IN A

4139
03:39:09,951 --> 03:39:11,219
SINGLE CHAIN VARIABLE FRAGMENT

4140
03:39:11,219 --> 03:39:12,920
FORMAT WHERE WE TAKE THE HEAVY

4141
03:39:12,920 --> 03:39:15,523
CHAIN AND LIGHT CHAINS OF THE

4142
03:39:15,523 --> 03:39:18,659
ANTIBODIES AND TETHER TOGETHER

4143
03:39:18,659 --> 03:39:20,361
WITH GLYCINE SERINE LINKER

4144
03:39:20,361 --> 03:39:24,198
RESULTING IN EACH YEAST CELL

4145
03:39:24,198 --> 03:39:25,600
DISPLAYING A DIFFERENT MUTANT,

4146
03:39:25,600 --> 03:39:27,502
WE CAN DETERMINE HOW THEY CAN

4147
03:39:27,502 --> 03:39:30,772
BIND TO HIV OMS BY LABELING THE

4148
03:39:30,772 --> 03:39:34,042
CELL SURFACE AND OMS LIGAND OF

4149
03:39:34,042 --> 03:39:37,145
CHOICE, THIS ALLOWS US TO

4150
03:39:37,145 --> 03:39:43,551
LEVERAGE FLUORESCENCE ACTIVATED

4151
03:39:43,551 --> 03:39:43,818
 ACTIVATED

4152
03:39:43,818 --> 03:39:46,320
CELL SORTING, WE CAN RUN THIS

4153
03:39:46,320 --> 03:39:48,389
YEAST CELL THROUGH A CYTOMETER

4154
03:39:48,389 --> 03:39:52,260
AND SORT INTO BINS BASED ON THE

4155
03:39:52,260 --> 03:39:53,094
BINDING FLUORESCENCE INTENSITY.

4156
03:39:53,094 --> 03:39:55,396
IF WE'RE TAKING A POOL OF YEAST

4157
03:39:55,396 --> 03:39:57,265
CELLS EXPRESSING A DIFFERENT

4158
03:39:57,265 --> 03:40:00,568
VARIANT ON CELL SURFACE WITH

4159
03:40:00,568 --> 03:40:02,403
DIFFERENT BINDING AFFINITIES AS

4160
03:40:02,403 --> 03:40:04,705
YOU TITRATE UP YOU'LL SEE THE

4161
03:40:04,705 --> 03:40:06,674
ONES WITH HIGHER BINDING

4162
03:40:06,674 --> 03:40:08,509
AFFINITY WILL TITRATE INTO BINS

4163
03:40:08,509 --> 03:40:11,579
3 AND 4 AT MUCH QUICKER RATE.

4164
03:40:11,579 --> 03:40:16,317
ONCE WE DO THE ASSAY WE CAN

4165
03:40:16,317 --> 03:40:19,053
EXACT THE DNA, DEEP SEQUENCE

4166
03:40:19,053 --> 03:40:20,154
VARIANTS, AND FROM THERE WE'LL

4167
03:40:20,154 --> 03:40:22,323
BE ABLE TO QUANTIFY AT PER

4168
03:40:22,323 --> 03:40:23,858
VARIANT LEVEL BINDING AT EACH

4169
03:40:23,858 --> 03:40:25,726
LIGAND CONCENTRATION WHICH AFTER

4170
03:40:25,726 --> 03:40:26,360
RUNNING COMPUTATIONAL ANALYSIS

4171
03:40:26,360 --> 03:40:30,231
CAN GET US TO VARIANT-SPECIFIC

4172
03:40:30,231 --> 03:40:32,066
TITRATION BINDING CURVES.

4173
03:40:32,066 --> 03:40:36,337
SO TO THIS EFFECT I'VE RUN THE

4174
03:40:36,337 --> 03:40:38,973
IOMA SINGLE MUTANT SCAN AND THIS

4175
03:40:38,973 --> 03:40:40,108
ASSAY WITH IOMA ANTIBODY, WITH

4176
03:40:40,108 --> 03:40:41,909
TWO LIGANDS.

4177
03:40:41,909 --> 03:40:47,181
I'VE DONE THE 4, 2, 6, A NATIVE

4178
03:40:47,181 --> 03:40:51,018
PROTEIN AND IgG 2 IMMUNOAGAIN

4179
03:40:51,018 --> 03:40:53,521
FROM 4, 2, 6C, THE READOUT WE

4180
03:40:53,521 --> 03:40:54,989
MIGHT EXPECT LOOKING AT A HEAT

4181
03:40:54,989 --> 03:41:01,262
MAP, CHANGE IN  BINDING AFFINIT,

4182
03:41:01,262 --> 03:41:03,331
AT THE HEAVY A AND LIGHT RAIN,

4183
03:41:03,331 --> 03:41:05,399
ON THE Y-AXIS WE'RE LOOKING AT

4184
03:41:05,399 --> 03:41:08,035
ALL MUTATIONS WE CAN MUTATE AT

4185
03:41:08,035 --> 03:41:14,275
THAT SITE, WHERE QUITE  SQUARES

4186
03:41:14,275 --> 03:41:15,810
WITH Xs WITH WILDTYPE RESIDUE.

4187
03:41:15,810 --> 03:41:18,312
IF WE LOOK AT THE HEAVY CHAIN,

4188
03:41:18,312 --> 03:41:20,081
THESE POSITIONS WERE CYSTEINE IS

4189
03:41:20,081 --> 03:41:22,049
THE WILD TYPE RESIDUE ANYTHING

4190
03:41:22,049 --> 03:41:26,287
ELSE YOU MUTATE THAT RESIDUE TO

4191
03:41:26,287 --> 03:41:27,188
WILL TANK BINDING AFFINITY,

4192
03:41:27,188 --> 03:41:33,728
INFERRING WE MIGHT BE DISRUPTING

4193
03:41:33,728 --> 03:41:36,597
A DISULFIDE BOND, FOR INSTANCE,

4194
03:41:36,597 --> 03:41:41,002
SEEING A LOWER BINDING AFFINITY.

4195
03:41:41,002 --> 03:41:42,837
WHAT WE'RE LOOKING AT HERE IS

4196
03:41:42,837 --> 03:41:45,339
TOP-DOWN VIEW OF THE BINDING

4197
03:41:45,339 --> 03:41:47,441
INTERFACE THAT IOMA USES TO

4198
03:41:47,441 --> 03:41:49,177
CONTACT OMS, THIS IS WHAT YOU

4199
03:41:49,177 --> 03:41:52,246
WOULD SEE FLYING AT YOU, WHERE

4200
03:41:52,246 --> 03:41:55,316
I'VE OUTLINED THE KEY RESIDUES

4201
03:41:55,316 --> 03:41:56,751
THAT CONTACT THE GP120 IN BLACK.

4202
03:41:56,751 --> 03:41:58,753
AND AS YOU MIGHT GLEAN FROM JUST

4203
03:41:58,753 --> 03:42:01,556
LOOKING AT THE HEAT MAP MOST OF

4204
03:42:01,556 --> 03:42:04,458
THESE MUTATIONS ARE NEGATIVE.

4205
03:42:04,458 --> 03:42:05,560
IF WE'RE LOOKING -- OR ON

4206
03:42:05,560 --> 03:42:06,661
AVERAGE, NOT NEGATIVE.

4207
03:42:06,661 --> 03:42:08,829
IF WE LOOK AT MAXIMUM BENEFICIAL

4208
03:42:08,829 --> 03:42:10,731
EFFECTS OF MUTATIONS WE START TO

4209
03:42:10,731 --> 03:42:11,632
SEE INTERESTING THINGS.

4210
03:42:11,632 --> 03:42:13,334
AROUND THE PERIPHERY OF THAT

4211
03:42:13,334 --> 03:42:16,337
CORE BINDING INTERFACE WE'RE

4212
03:42:16,337 --> 03:42:17,104
STARTING TO SEE COMPLEMENTARY

4213
03:42:17,104 --> 03:42:22,343
DETERMINING  LOOPS WITH MUTATIOS

4214
03:42:22,343 --> 03:42:24,745
WHICH BIND AFFINITY, EXCITING TO

4215
03:42:24,745 --> 03:42:24,946
SEE.

4216
03:42:24,946 --> 03:42:26,280
I WANT TO HIGHLIGHT EACH OF

4217
03:42:26,280 --> 03:42:29,350
THESE TILES IN A HEAT MAP IS A

4218
03:42:29,350 --> 03:42:32,286
TITRATION SERIES WITH PROTEIN

4219
03:42:32,286 --> 03:42:34,188
MUTANT, HIGHEST AFFINITY

4220
03:42:34,188 --> 03:42:37,358
ENHANCING MUTATION SERINE 100

4221
03:42:37,358 --> 03:42:40,328
WHEN WE MUTATE WE CAN INCREASE

4222
03:42:40,328 --> 03:42:42,630
BY ABOUT TEN-FOLD OR SO.

4223
03:42:42,630 --> 03:42:45,199
WHEN I LOOK AT THE STRUCTURE

4224
03:42:45,199 --> 03:42:47,835
WITH MODELING I CAN SEE I'M NOT

4225
03:42:47,835 --> 03:42:52,373
ABLE TO SEE ANY SIDE CHAINS TO

4226
03:42:52,373 --> 03:42:55,309
THAT MIGHT ACCOUNT FOR INCREASED

4227
03:42:55,309 --> 03:42:58,045
BINDING AFFINITY I'M SEEING A

4228
03:42:58,045 --> 03:42:59,380
STABILIZING EFFECT WITH LYSINE

4229
03:42:59,380 --> 03:43:01,015
28 MAY BE FORMING A SALT BRIDGE

4230
03:43:01,015 --> 03:43:03,484
BUT I HAVE TO RUN MORE ASSAYS TO

4231
03:43:03,484 --> 03:43:04,552
CHASE THAT DOWN AND CONFIRM.

4232
03:43:04,552 --> 03:43:07,555
IT GOES TO SHOW YOU CAN GO FROM

4233
03:43:07,555 --> 03:43:12,360
LARGE AERIAL VIEW OF THE HEAT

4234
03:43:12,360 --> 03:43:16,430
MAP LEVEL TO MORE OF A DISTINCT

4235
03:43:16,430 --> 03:43:17,298
MOLECULAR CONTACT LEVEL.

4236
03:43:17,298 --> 03:43:22,103
I'VE ALSO RUN THIS WITH THE

4237
03:43:22,103 --> 03:43:29,744
IgG 2 MUTAGEN, WE'RE SEEING

4238
03:43:29,744 --> 03:43:31,012
SIMILAR EFFECTS, BUT THE KEY

4239
03:43:31,012 --> 03:43:32,813
IDEA IS THERE'S A LOT OF

4240
03:43:32,813 --> 03:43:34,348
DIFFERENCES AS WELL AND YOU

4241
03:43:34,348 --> 03:43:35,449
WOULD BE ABSOLUTELY CORRECT.

4242
03:43:35,449 --> 03:43:38,819
I DECIDED TO HONE IN ON ONE

4243
03:43:38,819 --> 03:43:39,887
DIFFERENTIAL MUTATION THAT

4244
03:43:39,887 --> 03:43:43,557
LYSINE 58 HERE IN THE CDRH 2

4245
03:43:43,557 --> 03:43:43,858
LOOP.

4246
03:43:43,858 --> 03:43:47,228
WHEN WE MAP THE DIFFERENCE IN

4247
03:43:47,228 --> 03:43:49,096
BINDING AFFINITY WE CAN SEE

4248
03:43:49,096 --> 03:43:50,431
THAT'S MOSTLY A 1:1 EFFECT BUT

4249
03:43:50,431 --> 03:43:52,733
THERE ARE QUADRANTS WHERE WE'RE

4250
03:43:52,733 --> 03:43:53,768
SEEING DIFFERENTIAL MUTATIONAL

4251
03:43:53,768 --> 03:43:55,603
EFFECTS, AND THAT, AGAIN, LYSINE

4252
03:43:55,603 --> 03:43:57,571
58 AND HEAVY CHAIN IS SITTING IN

4253
03:43:57,571 --> 03:44:06,047
THE AREA WITH INCREASED

4254
03:44:06,047 --> 03:44:06,614
AFFINITY.

4255
03:44:06,614 --> 03:44:10,651
THAT MUTATION IS JAM PACKED INTO

4256
03:44:10,651 --> 03:44:12,653
WHERE ALL THE MUTATIONS FROM

4257
03:44:12,653 --> 03:44:13,954
426C WERE MADE.

4258
03:44:13,954 --> 03:44:15,589
THAT'S EPRESIDENT-ELECTED IN THE

4259
03:44:15,589 --> 03:44:24,098
BINDING  CURVES THAT CORRELATE.

4260
03:44:24,098 --> 03:44:24,899
MY SUMMARY, TAKEHOME MESSAGE

4261
03:44:24,899 --> 03:44:27,001
I'VE STARTED TO BUILD A SYSTEM

4262
03:44:27,001 --> 03:44:28,602
THAT CAN PROBE THESE FUNCTIONAL

4263
03:44:28,602 --> 03:44:32,239
FLEXIBILITIES AND CONSTRAINTS IN

4264
03:44:32,239 --> 03:44:35,843
A HOLISTIC MANNER AND I HOPE

4265
03:44:35,843 --> 03:44:40,181
I'VE CONVINCED YOU MUTATIONS

4266
03:44:40,181 --> 03:44:46,020
SHOW DIFFERING EFFECT, WHEN YOU

4267
03:44:46,020 --> 03:44:47,988
LOOK AT THIS MUTATIONAL PATHWAYS

4268
03:44:47,988 --> 03:44:49,256
CAN BE INDUCED.

4269
03:44:49,256 --> 03:44:53,227
NEXT STEPS, I WANT TO EXPAND TO

4270
03:44:53,227 --> 03:44:56,163
VRC-01 AND OTHER ANTIBODIES, THE

4271
03:44:56,163 --> 03:44:57,598
LIGAND PANEL THAT I CAN WORK

4272
03:44:57,598 --> 03:45:01,769
WITH SO I CAN SEE HOW MUTATIONS

4273
03:45:01,769 --> 03:45:03,537
AFFECT RECOGNITION ON A GLOBAL

4274
03:45:03,537 --> 03:45:08,542
HIV DIVERSITY LEVEL AND SOME

4275
03:45:08,542 --> 03:45:10,745
OTHER IMMUNOGENS AND AUTOLOGOUS

4276
03:45:10,745 --> 03:45:13,581
PROTEINS TO FLESH OUT SURFACE

4277
03:45:13,581 --> 03:45:15,649
GLYCOSYLATION CAN BE OVERCOME

4278
03:45:15,649 --> 03:45:16,450
WITH DIFFERENT MUTATIONS.

4279
03:45:16,450 --> 03:45:18,719
I'D LIKE TO ACKNOWLEDGE MY LAB,

4280
03:45:18,719 --> 03:45:20,488
HELPFUL GETTING THIS OFF THE

4281
03:45:20,488 --> 03:45:21,989
GROUND, ESPECIALLY THE BJORKMAN

4282
03:45:21,989 --> 03:45:23,958
GROUP WHO PROVIDED REAGENTS AND

4283
03:45:23,958 --> 03:45:25,693
SCIENTIFIC EXPERTISE AS WELL AS

4284
03:45:25,693 --> 03:45:27,228
CHEETAH GROUP FOR ALLOWING ME TO

4285
03:45:27,228 --> 03:45:28,562
COME AND TELL MY STORY.

4286
03:45:28,562 --> 03:45:37,905
THANK YOU. 

4287
03:45:37,905 --> 03:45:40,474
>> ANY QUESTIONS FOR
JAMIN?

4288
03:45:40,474 --> 03:45:46,080
NOBODY? 

4289
03:45:46,080 --> 03:45:46,714
>> ALL RIGHT.

4290
03:45:46,714 --> 03:45:48,549
THANK YOU.

4291
03:45:48,549 --> 03:45:50,184
>> NICE JOB, JAMIN.

4292
03:45:50,184 --> 03:45:50,551
THANK YOU.

4293
03:45:50,551 --> 03:45:53,487
[APPLAUSE]

4294
03:45:53,487 --> 03:45:55,890
NEXT SPEAKER WILL BE RUTH

4295
03:45:55,890 --> 03:45:58,959
PARSONS, DUKE UNIVERSITY.

4296
03:45:58,959 --> 03:46:04,999
 

4297
03:46:04,999 --> 03:46:15,042
 

4298
03:46:36,997 --> 03:46:40,134
 

4299
03:46:40,134 --> 03:46:42,002
>> HEY, EVERYBODY.

4300
03:46:42,002 --> 03:46:46,040
I'M RUTH PARSONS, A FOURTH YEAR

4301
03:46:46,040 --> 03:46:47,007
GRADUATE STUDENT, AT DUKE

4302
03:46:47,007 --> 03:46:47,408
UNIVERSITY.

4303
03:46:47,408 --> 03:46:51,512
TODAY I'M GOING TO TELL YOU

4304
03:46:51,512 --> 03:46:57,651
ABOUT CONFIRMATIONAL  VARIATION.

4305
03:46:57,651 --> 03:46:59,386
CD4 BINDING IS IMPORTANT AT THE

4306
03:46:59,386 --> 03:47:01,121
BEGINNING OF THE TRANSITION OF

4307
03:47:01,121 --> 03:47:03,557
THE ENVELOPE FROM PRE-FUSION TO

4308
03:47:03,557 --> 03:47:03,991
POST-FUSION.

4309
03:47:03,991 --> 03:47:05,392
THERE'S A PRIMARY BINDING SITE

4310
03:47:05,392 --> 03:47:08,395
AND SECONDARY BINDING SITE FOR

4311
03:47:08,395 --> 03:47:08,562
CD4.

4312
03:47:08,562 --> 03:47:12,433
SHOWN IN BLUE IS PRIMARY BINDING

4313
03:47:12,433 --> 03:47:12,633
SITE.

4314
03:47:12,633 --> 03:47:15,369
ON ON THE ADJACENT GP120 THERE'S

4315
03:47:15,369 --> 03:47:16,570
A SECONDARY BINDING SITE.

4316
03:47:16,570 --> 03:47:19,406
THIS IS IMPORTANT FOR INITIAL

4317
03:47:19,406 --> 03:47:20,508
CONTACT OF CD4.

4318
03:47:20,508 --> 03:47:22,843
THERE ARE MANY bNAbS THAT

4319
03:47:22,843 --> 03:47:26,881
BIND TO CD4 AS WE'VE BEEN

4320
03:47:26,881 --> 03:47:27,181
DISCUSSING.

4321
03:47:27,181 --> 03:47:29,817
AND THERE ARE A FEW THAT BIND TO

4322
03:47:29,817 --> 03:47:31,352
BOTH THE PRIMARY BINDING SITE

4323
03:47:31,352 --> 03:47:33,521
AND THE SECONDARY BINDING SITE.

4324
03:47:33,521 --> 03:47:38,259
AND THAT'S WHAT IS OF INTEREST

4325
03:47:38,259 --> 03:47:40,528
IN MY TALK TODAY.

4326
03:47:40,528 --> 03:47:43,130
THERE ARE OBSTACLES FOR BROADLY

4327
03:47:43,130 --> 03:47:43,898
NEUTRALIZING ANTIBODIES BEING

4328
03:47:43,898 --> 03:47:48,302
ABLE TO BIND TO THE CD4 BINDING

4329
03:47:48,302 --> 03:47:50,604
SITE ONE AND TWO.

4330
03:47:50,604 --> 03:47:55,609
THERE'S VERY -- IT'S A VERY

4331
03:47:55,609 --> 03:47:57,378
VARIED SITE BETWEEN TWO

4332
03:47:57,378 --> 03:47:59,680
PROTOMERS, LONG LOOPS, GLYCANS

4333
03:47:59,680 --> 03:48:00,981
SHIELDING THE REGION, AND

4334
03:48:00,981 --> 03:48:04,418
THERE'S ALSO FLEXIBILITY BETWEEN

4335
03:48:04,418 --> 03:48:07,588
THE TWO ENV MONOMERS.

4336
03:48:07,588 --> 03:48:10,624
THAT BINDING SITE IS BETWEEN TWO

4337
03:48:10,624 --> 03:48:11,158
MONOMERS, FLEXIBILITY IN

4338
03:48:11,158 --> 03:48:11,825
MOVEMENT BETWEEN THEM.

4339
03:48:11,825 --> 03:48:14,528
OUR GOAL IS TO UNDERSTAND HOW

4340
03:48:14,528 --> 03:48:17,965
THE CD4 BINDING SITE bNAbS

4341
03:48:17,965 --> 03:48:20,568
ACCESS THE CD 4 BINDING SITE AND

4342
03:48:20,568 --> 03:48:25,072
CD4 BINDING SITE 2.

4343
03:48:25,072 --> 03:48:28,909
OUR GROUP STUDIED ENV

4344
03:48:28,909 --> 03:48:31,078
FLEXIBILITY, THAT IS BETWEEN TWO

4345
03:48:31,078 --> 03:48:31,412
PROTOMERS.

4346
03:48:31,412 --> 03:48:35,149
WE'VE FOUND DIFFERENCES IN

4347
03:48:35,149 --> 03:48:36,550
FLEXIBILITY BETWEEN DIFFERENT

4348
03:48:36,550 --> 03:48:39,653
SEQUENCES AND ENVELOPES.

4349
03:48:39,653 --> 03:48:43,190
SO HERE SHOWN A BG 505 SOSIP,

4350
03:48:43,190 --> 03:48:45,626
TIGHTLY CLOSED ENVELOPE, AND

4351
03:48:45,626 --> 03:48:47,061
THEN 1059 SOSIP, COMPLETELY

4352
03:48:47,061 --> 03:48:47,695
OPEN.

4353
03:48:47,695 --> 03:48:50,965
IT'S ALSO MORE FLEXIBLE.

4354
03:48:50,965 --> 03:48:53,500
AND A CONSEQUENCE OF THIS WAS

4355
03:48:53,500 --> 03:48:56,904
FOUND TO BE DECREASED BINDING TO

4356
03:48:56,904 --> 03:48:58,205
THE CD4 BINDING SITE ANTIBODIES.

4357
03:48:58,205 --> 03:49:00,841
THEY BIND FINE TO JUST THE GP120

4358
03:49:00,841 --> 03:49:02,810
BUT IN THE TRIMERIC FORM THEY

4359
03:49:02,810 --> 03:49:05,312
DON'T BIND AS WELL.

4360
03:49:05,312 --> 03:49:09,717
AND SO WE BELIEVE THAT'S DUE TO

4361
03:49:09,717 --> 03:49:12,286
THE FLEXIBILITY.

4362
03:49:12,286 --> 03:49:14,088
SO, THERE ARE DIFFERENT WAYS

4363
03:49:14,088 --> 03:49:15,155
THAT DIFFERENT bNAbS HAVE

4364
03:49:15,155 --> 03:49:18,459
FIGURED OUT HOW TO GET TO THE

4365
03:49:18,459 --> 03:49:28,869
SECONDARY BINDING SITE.

4366
03:49:29,803 --> 03:49:32,006
LIKE ONE EXAMPLE, VRC 03 HAS

4367
03:49:32,006 --> 03:49:33,874
EXTENDED FR 3 REGION HERE, THAT

4368
03:49:33,874 --> 03:49:37,611
ALLOWS US TO REACH TO THE

4369
03:49:37,611 --> 03:49:39,013
SECONDARY BINDING SITE.

4370
03:49:39,013 --> 03:49:42,349
VRC CH 31 AS EXTENDED REGION

4371
03:49:42,349 --> 03:49:46,353
WHICH ALLOWS IT TO EXTEND TO

4372
03:49:46,353 --> 03:49:48,455
THAT REGION.

4373
03:49:48,455 --> 03:49:51,525
AND THEN ANOTHER ANTIBODY THAT

4374
03:49:51,525 --> 03:49:54,595
WE'RE INTERESTED IN CR 103

4375
03:49:54,595 --> 03:49:59,166
DOESN'T HAVE EXTENDED LOOPS.

4376
03:49:59,166 --> 03:50:00,701
DIFFERENT ANGLE OF APPROACH,

4377
03:50:00,701 --> 03:50:02,903
20-DEGREE DIFFERENT IN ANGLE,

4378
03:50:02,903 --> 03:50:04,405
THAT ALLOWS THIS FRAMEWORK

4379
03:50:04,405 --> 03:50:06,040
REGION 3 LOOP TO GET CLOSE

4380
03:50:06,040 --> 03:50:09,543
ENOUGH TO THE SECONDARY BINDING

4381
03:50:09,543 --> 03:50:11,445
SITE TO INTERACT.

4382
03:50:11,445 --> 03:50:20,320
SO WE'RE INTERESTED IN THIS CH

4383
03:50:20,320 --> 03:50:21,955
103 bNAb AND WANT TO

4384
03:50:21,955 --> 03:50:22,856
UNDERSTAND HOW THE RESIDUES

4385
03:50:22,856 --> 03:50:24,091
EVOLVED TO BIND TO THIS SITE.

4386
03:50:24,091 --> 03:50:29,063
WE'RE INTERESTED IN TWO RESIDUES

4387
03:50:29,063 --> 03:50:30,664
IN THE FRAMEWORK REGION 3 THAT

4388
03:50:30,664 --> 03:50:35,102
EARLY ON IN THE ANTIBODY LINEAGE

4389
03:50:35,102 --> 03:50:41,508
DEVELOPMENT THEY MUTATED TO

4390
03:50:41,508 --> 03:50:43,477
BECOME MORE AN-IONIC, MAINTAINED

4391
03:50:43,477 --> 03:50:45,913
THROUGH THE LINEAGE AND MATURE

4392
03:50:45,913 --> 03:50:47,047
CH 103.

4393
03:50:47,047 --> 03:50:48,716
AS I MENTIONED THEY ARE SITUATED

4394
03:50:48,716 --> 03:50:50,250
IN REGION 3 WHICH IS WHAT

4395
03:50:50,250 --> 03:50:54,755
INTERACTS WITH THE CD4 BINDING

4396
03:50:54,755 --> 03:50:59,493
SITE 2, CDRH 3, INTERACTING WITH

4397
03:50:59,493 --> 03:51:00,994
BINDING SITE 1.

4398
03:51:00,994 --> 03:51:02,830
AND SO THE UNMUTATED COME AN

4399
03:51:02,830 --> 03:51:04,031
ANCESTOR, IF YOU MUTATE THESE

4400
03:51:04,031 --> 03:51:06,233
TWO RESIDUES BACK TO THE

4401
03:51:06,233 --> 03:51:08,435
UNMUTATED COMMON ANCESTOR AT

4402
03:51:08,435 --> 03:51:12,272
POSITION 75 AND 76, THAT

4403
03:51:12,272 --> 03:51:12,873
DECREASES NEUTRALIZATION BY

4404
03:51:12,873 --> 03:51:16,110
THESE ANTIBODIES AS WELL AS

4405
03:51:16,110 --> 03:51:25,919
BINDING TO THE  THE ENV.

4406
03:51:25,919 --> 03:51:28,355
THE UNMUTATED COMMON ANCESTOR

4407
03:51:28,355 --> 03:51:31,825
REVERTED FORM ALL REFER TO THAT

4408
03:51:31,825 --> 03:51:33,694
AS THE KN MUTANT.

4409
03:51:33,694 --> 03:51:35,662
SO WE COLLECTED DATASETS ON BOTH

4410
03:51:35,662 --> 03:51:38,732
OF THESE AND GOT MULTIPLE

4411
03:51:38,732 --> 03:51:40,267
STRUCTURES WITH DIFFERENT

4412
03:51:40,267 --> 03:51:41,468
NUMBERS OF ENVs BOUND.

4413
03:51:41,468 --> 03:51:44,538
MANY OF YOU ARE FAMILIAR WITH

4414
03:51:44,538 --> 03:51:48,142
THE CryoEM DATA PROCESSING

4415
03:51:48,142 --> 03:51:50,110
WORKFLOW, COLLECTING

4416
03:51:50,110 --> 03:51:51,111
MICROGRAPHS, TAKING PARTICLES,

4417
03:51:51,111 --> 03:51:57,718
DOING  2D CLASS AVERAGES, WE GET

4418
03:51:57,718 --> 03:51:59,620
RID OF BAD PARTICLES AND

4419
03:51:59,620 --> 03:52:01,255
CLASSES, THEN WE CAN GET

4420
03:52:01,255 --> 03:52:03,724
MULTIPLE STRUCTURES OUT OF ONE

4421
03:52:03,724 --> 03:52:03,957
DATASET.

4422
03:52:03,957 --> 03:52:06,160
SO FOR MY TWO DATASETS I'M

4423
03:52:06,160 --> 03:52:10,831
TALKING ABOUT NOW, WILD TYPE AND

4424
03:52:10,831 --> 03:52:15,736
REVERTED, I HAVE STRUCTURES OF 1

4425
03:52:15,736 --> 03:52:20,407
FAB BOUND, 2 BOUND, THEN NO FAB

4426
03:52:20,407 --> 03:52:22,476
FOR THE KN MUTANT.

4427
03:52:22,476 --> 03:52:23,877
NOW ZOOMING INTO THE BINDING

4428
03:52:23,877 --> 03:52:31,318
SITE TO SEE WHAT'S GOING ON, AT

4429
03:52:31,318 --> 03:52:32,986
THE POSITION 75, THIS INTERACTS

4430
03:52:32,986 --> 03:52:34,922
WITH THIS ARGININE HERE SO

4431
03:52:34,922 --> 03:52:36,890
THERE'S AN EXTENSIVE LIKE

4432
03:52:36,890 --> 03:52:39,193
HYDROGEN BONDING NETWORK THAT'S

4433
03:52:39,193 --> 03:52:43,063
CREATED HERE SO THESE RESIDUES

4434
03:52:43,063 --> 03:52:46,200
ARE SITUATED CLOSE TO THE

4435
03:52:46,200 --> 03:52:46,633
ARGININE.

4436
03:52:46,633 --> 03:52:49,570
THEN IN CONTRAST THE LYSINE THAT

4437
03:52:49,570 --> 03:52:52,940
THIS 75 HAS BEEN MUTATED TO IS

4438
03:52:52,940 --> 03:52:59,313
NOW SITUATED TOO FAR TO BE

4439
03:52:59,313 --> 03:53:00,747
HYDROGEN BONDING.

4440
03:53:00,747 --> 03:53:02,749
POSITION 76, IN THE WILD TYPE,

4441
03:53:02,749 --> 03:53:06,220
IT IS SITUATED CLOSE ENOUGH TO

4442
03:53:06,220 --> 03:53:08,422
HAVE ELECTROSTATIC INTERACTIONS

4443
03:53:08,422 --> 03:53:10,490
WITH LYSINE, IN THE KN MUTANT

4444
03:53:10,490 --> 03:53:12,259
THIS LYSINE IS NOW SITUATED TOO

4445
03:53:12,259 --> 03:53:14,862
FAR AWAY TO BE INTERACTING.

4446
03:53:14,862 --> 03:53:19,132
AND THERE'S ALSO NOT VERY GOOD

4447
03:53:19,132 --> 03:53:20,534
DENSITY INDICATING THIS IS NOT

4448
03:53:20,534 --> 03:53:23,937
CREATING A STABLE INTERACTION AT

4449
03:53:23,937 --> 03:53:26,974
THIS PORTION OF THE CD4 BINDING

4450
03:53:26,974 --> 03:53:30,611
SITE, THIS LOWER PORTION OF THE

4451
03:53:30,611 --> 03:53:32,479
CD4 BINDING SITE 2 IS CREATING

4452
03:53:32,479 --> 03:53:36,083
MORE OF THE DIFFERENCE IN THE

4453
03:53:36,083 --> 03:53:37,851
BINDING BETWEEN THOSE TWO

4454
03:53:37,851 --> 03:53:41,421
MUTANTS.

4455
03:53:41,421 --> 03:53:43,590
AND I ALSO TOOK A LOOK AT THE

4456
03:53:43,590 --> 03:53:44,892
PRIMARY BINDING SITE TO SEE IF

4457
03:53:44,892 --> 03:53:47,027
THE DIFFERENCES IN THE SECONDARY

4458
03:53:47,027 --> 03:53:48,428
BINDING SITE WOULD CAUSE ANY

4459
03:53:48,428 --> 03:53:49,963
CHANGES IN THE PRIMARY BINDING

4460
03:53:49,963 --> 03:53:50,397
SITE.

4461
03:53:50,397 --> 03:53:52,132
AND I DIDN'T SEE ANY DIFFERENCES

4462
03:53:52,132 --> 03:53:55,335
SO IT SEEMS TO BE BINDING THE

4463
03:53:55,335 --> 03:53:57,437
WILD TYPE AND KN SEEM TO BE

4464
03:53:57,437 --> 03:54:00,040
BINDING WITH EACH OTHER TO THE

4465
03:54:00,040 --> 03:54:01,909
PRIMARY BINDING SITE SO ALL

4466
03:54:01,909 --> 03:54:03,577
THESE INTERACTIONS I'M SHOWING

4467
03:54:03,577 --> 03:54:08,916
ARE JUST THE SAME BETWEEN THE

4468
03:54:08,916 --> 03:54:09,316
TWO.

4469
03:54:09,316 --> 03:54:11,518
SO, TO FURTHER LOOK AT THE

4470
03:54:11,518 --> 03:54:13,954
FLEXIBILITY IN THESE STRUCTURES,

4471
03:54:13,954 --> 03:54:15,589
I USED 3D VARIABILITY ANALYSIS

4472
03:54:15,589 --> 03:54:17,257
SO THIS TAKES INDIVIDUAL

4473
03:54:17,257 --> 03:54:18,692
PARTICLE IMAGES AND DOES

4474
03:54:18,692 --> 03:54:20,127
PRINCIPAL COMPONENTS ANALYSIS TO

4475
03:54:20,127 --> 03:54:21,728
GIVE A CONTINUOUS FAMILY OF 3D

4476
03:54:21,728 --> 03:54:22,963
STRUCTURES, AND THAT'S WHAT I

4477
03:54:22,963 --> 03:54:25,632
WAS SHOWING, THAT'S WHAT WE DID

4478
03:54:25,632 --> 03:54:27,034
IN THOSE STRUCTURES OF ENVELOPES

4479
03:54:27,034 --> 03:54:30,437
TO SHOW DIFFERENT ENVELOPES HAVE

4480
03:54:30,437 --> 03:54:30,904
DIFFERENT FLEXIBILITY.

4481
03:54:30,904 --> 03:54:33,073
I DID THAT WITH THESE STRUCTURES

4482
03:54:33,073 --> 03:54:34,942
AS WELL.

4483
03:54:34,942 --> 03:54:38,178
AND THESE ARE THE MOVIES THAT

4484
03:54:38,178 --> 03:54:41,915
RESULTED FROM DOING 3D ANALYSIS,

4485
03:54:41,915 --> 03:54:43,583
IN GENERAL THE WILD TYPE FORM

4486
03:54:43,583 --> 03:54:45,886
SEEM TO BE LESS FLEXIBLE,

4487
03:54:45,886 --> 03:54:47,921
ESPECIALLY AT THIS PROTOMER

4488
03:54:47,921 --> 03:54:49,690
SITUATED NEXT TO THE FAB.

4489
03:54:49,690 --> 03:54:52,793
AND THEN IN THE BINDING SITE YOU

4490
03:54:52,793 --> 03:54:54,828
CAN CLEARLY SEE IN ALL OF THE

4491
03:54:54,828 --> 03:54:56,863
FRAMES THERE IS DENSITY AROUND

4492
03:54:56,863 --> 03:54:59,232
THIS LYSINE, THAT I MENTIONED

4493
03:54:59,232 --> 03:55:00,500
WAS INTERACTING EXTENSIVELY BUT

4494
03:55:00,500 --> 03:55:06,807
THAT IS NOT REALLY PRESENT IN

4495
03:55:06,807 --> 03:55:07,674
THE KN MUTANT.

4496
03:55:07,674 --> 03:55:09,776
AND ALSO YOU CAN SEE THIS

4497
03:55:09,776 --> 03:55:10,944
PROTOMER IS JUST MOVING MUCH

4498
03:55:10,944 --> 03:55:12,479
MORE AND NOT CREATING A STABLE

4499
03:55:12,479 --> 03:55:14,314
INTERACTION AT THIS PORTION OF

4500
03:55:14,314 --> 03:55:16,783
THE INTERACTION.

4501
03:55:16,783 --> 03:55:20,120
OF THE BINDING SITE.

4502
03:55:20,120 --> 03:55:21,421
IN CONCLUSION, HIV-1 ENVELOPE

4503
03:55:21,421 --> 03:55:28,562
FOR BINDING SITE BETWEEN TWO

4504
03:55:28,562 --> 03:55:30,297
PROTOMERS PROTECTED BY

4505
03:55:30,297 --> 03:55:31,665
FLEXIBILITY, SOME bNAbS

4506
03:55:31,665 --> 03:55:33,800
INTERACT WITH PROTOMERS AT CD4

4507
03:55:33,800 --> 03:55:36,570
BINDING SITE USING MULTIPLE

4508
03:55:36,570 --> 03:55:43,677
MECHANISMS, AND CD103 LINEAGE

4509
03:55:43,677 --> 03:55:44,411
ACQUIRES, TWO MUTATIONS EARLY.

4510
03:55:44,411 --> 03:55:46,747
I WANT TO THANK MY LAB AND THE

4511
03:55:46,747 --> 03:55:49,116
FUNDING AND THE ORGANIZERS OF

4512
03:55:49,116 --> 03:55:49,983
THE THIS EVENT.

4513
03:55:49,983 --> 03:55:51,918
THANK YOU ALL VERY MUCH FOR

4514
03:55:51,918 --> 03:55:52,419
LISTENING.

4515
03:55:52,419 --> 03:56:00,293
[APPLAUSE] 

4516
03:56:00,293 --> 03:56:04,631
 

4517
03:56:04,631 --> 03:56:07,434
>> ANY QUESTIONS FOR RUTH?

4518
03:56:07,434 --> 03:56:10,737
 

4519
03:56:10,737 --> 03:56:13,473
QUIET GROUP THIS AFTERNOON.

4520
03:56:13,473 --> 03:56:15,108
 

4521
03:56:15,108 --> 03:56:15,909
ALL RIGHT.

4522
03:56:15,909 --> 03:56:18,612
WELL, FOR AN EXCELLENT

4523
03:56:18,612 --> 03:56:18,945
PRESENTATION.

4524
03:56:18,945 --> 03:56:23,450
[APPLAUSE]

4525
03:56:23,450 --> 03:56:29,456
NEXT AND FINAL SPEAKER IS MAMTA

4526
03:56:29,456 --> 03:56:30,157
SINGH FROM NATIONAL INSTITUTE OF

4527
03:56:30,157 --> 03:56:38,999
ALLERGY AND INFECTIOUS DISEASES 

4528
03:56:38,999 --> 03:56:49,042
 

4529
03:56:57,918 --> 03:57:01,922
 

4530
03:57:01,922 --> 03:57:05,492
>> GOOD AFTERNOON,
EVERYONE. 

4531
03:57:05,492 --> 03:57:08,395
I'M A POSTDOC FELLOW AT NIAID.

4532
03:57:08,395 --> 03:57:12,666
I'LL BE TALKING ABOUT THE

4533
03:57:12,666 --> 03:57:14,367
OPTIMIZATION OF VLP-FORMING

4534
03:57:14,367 --> 03:57:16,169
mRNA VACCINE BY

4535
03:57:16,169 --> 03:57:17,904
PROTEASE-MEDIATED GAG

4536
03:57:17,904 --> 03:57:18,205
PROCESSING.

4537
03:57:18,205 --> 03:57:19,606
SO, THE DEVELOPMENT OF THE

4538
03:57:19,606 --> 03:57:21,708
PROPHYLACTIC VACCINE REMAINS ONE

4539
03:57:21,708 --> 03:57:23,210
OF THE HIGHEST PRIORITIES IN THE

4540
03:57:23,210 --> 03:57:26,379
GLOBAL EFFORTS TO END THE

4541
03:57:26,379 --> 03:57:26,947
HIV/AIDS PANDEMIC.

4542
03:57:26,947 --> 03:57:30,617
AND THERE HAVE BEEN A WIDE

4543
03:57:30,617 --> 03:57:32,652
CONSENSUS THAT THE ELICITATION

4544
03:57:32,652 --> 03:57:34,087
OF BROADLY NEUTRALIZING

4545
03:57:34,087 --> 03:57:35,856
ANTIBODIES WHICH I TARGETED TO

4546
03:57:35,856 --> 03:57:37,591
THE ENVELOPE GLYCOPROTEIN IS THE

4547
03:57:37,591 --> 03:57:40,127
PRIMARY GOAL OF THE PROTECTIVE

4548
03:57:40,127 --> 03:57:40,427
VACCINE.

4549
03:57:40,427 --> 03:57:42,496
HOWEVER, THERE ARE SEVERAL KEY

4550
03:57:42,496 --> 03:57:45,999
REQUIREMENTS FOR THE ELICITATION

4551
03:57:45,999 --> 03:57:47,968
OF THESE bNAbS WHICH

4552
03:57:47,968 --> 03:57:51,605
BASICALLY REQUIRE THE ENVELOPE

4553
03:57:51,605 --> 03:58:00,647
GLYCOPROTEIN TO BE PRESENTED IN

4554
03:58:00,647 --> 03:58:01,715
ITS NATIVE CONFIGURATION.

4555
03:58:01,715 --> 03:58:05,485
THE ENVELOPE NEEDS TO BE

4556
03:58:05,485 --> 03:58:09,856
PRESENTED IN ITS NATIVE

4557
03:58:09,856 --> 03:58:17,831
CONFIGURATION TO RETAIN CORRECT

4558
03:58:17,831 --> 03:58:19,332
ANTI--GENICITY, TO STIMULATE

4559
03:58:19,332 --> 03:58:20,634
GERMINAL CENTER REACTION, WITH

4560
03:58:20,634 --> 03:58:24,204
EFFECTIVE ENGAGEMENT OF THE

4561
03:58:24,204 --> 03:58:27,307
PRECURSOR B CELLS FILED BY

4562
03:58:27,307 --> 03:58:29,609
MATURATION TOWARDS THE bNAb

4563
03:58:29,609 --> 03:58:29,910
DEVELOPMENT.

4564
03:58:29,910 --> 03:58:33,513
SO, TO ACHIEVE THESE GOALS IN

4565
03:58:33,513 --> 03:58:36,750
OUR LABORATORY WE DEVELOPED ON

4566
03:58:36,750 --> 03:58:38,752
THE GAG-BASED mRNA VACCINE

4567
03:58:38,752 --> 03:58:40,387
WHICH UTILIZES HOST

4568
03:58:40,387 --> 03:58:42,155
TRANSLATIONAL MACHINERY TO

4569
03:58:42,155 --> 03:58:43,924
EXPRESS VIRUS-LIKE PARTICLES AND

4570
03:58:43,924 --> 03:58:47,060
THE VACCINE WAS ADMINISTERED

4571
03:58:47,060 --> 03:58:48,728
USING UNIQUE IMMUNIZATION

4572
03:58:48,728 --> 03:58:51,131
REGIMEN, THAT INVOLVES PRIMING

4573
03:58:51,131 --> 03:58:52,332
WITH GERMLINE ENGAGER ENVELOPE

4574
03:58:52,332 --> 03:58:54,501
FOLLOWED BY INTENSIVE BOOSTING

4575
03:58:54,501 --> 03:58:56,169
WITH THE HETEROLOGOUS ENVELOPE

4576
03:58:56,169 --> 03:58:58,672
TO DRIVE THE AFFINITY MATURATION

4577
03:58:58,672 --> 03:59:00,774
TOWARDS bNAb DEVELOPMENT.

4578
03:59:00,774 --> 03:59:04,044
SO THE RESULTS OF THE STUDY WERE

4579
03:59:04,044 --> 03:59:07,013
REPORTED IN NATURE MEDICINE IN

4580
03:59:07,013 --> 03:59:08,415
2021.

4581
03:59:08,415 --> 03:59:14,020
IMPORTANTLY, ALL OF THE

4582
03:59:14,020 --> 03:59:15,755
VACCINATED MACAQUES INTRODUCED

4583
03:59:15,755 --> 03:59:16,756
TITERS AGAINST 11 OF THE 12

4584
03:59:16,756 --> 03:59:21,394
ISOLATES PART OF THE SO-CALLED

4585
03:59:21,394 --> 03:59:25,065
SMALL GLOBAL PANEL. 

4586
03:59:25,065 --> 03:59:29,336
EVEN THOUGH THE TITERS WERE LOW,

4587
03:59:29,336 --> 03:59:32,772
THE VACCINE INDUCED REMARKABLE

4588
03:59:32,772 --> 03:59:35,709
PROTECTION AGAINST REPEATED

4589
03:59:35,709 --> 03:59:38,845
MUCOSAL CHALLENGES WITH

4590
03:59:38,845 --> 03:59:40,847
HETEROLOGOUS NEUTRALIZED TITER

4591
03:59:40,847 --> 03:59:44,284
TO STRAIN THAT RADIATE WITH

4592
03:59:44,284 --> 03:59:46,553
CALCULATED RISK REDUCTION OF

4593
03:59:46,553 --> 03:59:47,721
79%.

4594
03:59:47,721 --> 03:59:52,092
THIS DATA SUGGESTED THE ENVELOPE

4595
03:59:52,092 --> 03:59:54,394
GAG VLP-FORMING VACCINE IS A

4596
03:59:54,394 --> 03:59:57,697
PROMISING APPROACH IN VACCINE

4597
03:59:57,697 --> 04:00:01,034
DEVELOPMENT, ROOM FOR FURTHER

4598
04:00:01,034 --> 04:00:01,334
IMPROVEMENT.

4599
04:00:01,334 --> 04:00:07,474
WITH THIS AIM IN THE STUDY WE

4600
04:00:07,474 --> 04:00:09,743
EXPLORED BENEFITS OF PRODUCING

4601
04:00:09,743 --> 04:00:14,381
MATURE RATHER THAN IMMATURE

4602
04:00:14,381 --> 04:00:19,686
VLPs TO DELIVER ENVELOPE

4603
04:00:19,686 --> 04:00:20,453
GLYCOPROTEIN.

4604
04:00:20,453 --> 04:00:22,122
THIS SHOWED RELATIVE TO GAG

4605
04:00:22,122 --> 04:00:23,757
LATTICE, IN CASE OF IMMATURE

4606
04:00:23,757 --> 04:00:26,660
PARTICLES IT HAS BEEN SEEN THAT

4607
04:00:26,660 --> 04:00:30,096
THE MATRIX DOMAIN OF THE GAG

4608
04:00:30,096 --> 04:00:33,400
INTERACTS WITH CYTOPLASTIC TAIL

4609
04:00:33,400 --> 04:00:34,834
OF THE ENVELOPE, THERE WERE

4610
04:00:34,834 --> 04:00:44,010
MOBILITY ACROSS THE ENTER

4611
04:00:44,010 --> 04:00:44,577
 INTERFACE.

4612
04:00:44,577 --> 04:00:47,113
IT IMPARTS MOBILITY TO THE

4613
04:00:47,113 --> 04:00:48,748
INCURRED GLYCOPROTEIN SO WE

4614
04:00:48,748 --> 04:00:50,483
SPECULATE THIS ENHANCED MOBILITY

4615
04:00:50,483 --> 04:00:53,920
OF ENVELOPE ON THE SURFACE OF

4616
04:00:53,920 --> 04:00:56,756
VIRUS-LIKE PARTICLES MAY IMPART

4617
04:00:56,756 --> 04:00:58,925
POTENTIAL IMMUNOLOGIC BENEFITS

4618
04:00:58,925 --> 04:01:04,064
IN TERMS OF ANTIGEN UPTAKE OR

4619
04:01:04,064 --> 04:01:06,032
PRESENTATION, MATURE VLPs MAY

4620
04:01:06,032 --> 04:01:09,436
ACT AS EFFECTIVE IMMUNOGEN

4621
04:01:09,436 --> 04:01:12,272
COMPARED TO IMMATURE

4622
04:01:12,272 --> 04:01:14,674
COUNTERPARTS.

4623
04:01:14,674 --> 04:01:16,543
WE INTRODUCED WILD PROTEASE OF A

4624
04:01:16,543 --> 04:01:26,953
THIRD COME  COMPLEMENT. 

4625
04:01:28,621 --> 04:01:31,291
WE HYPOTHESIZED THIS WILL

4626
04:01:31,291 --> 04:01:32,459
ENHANCE MOBILITY OF ENVELOPE,

4627
04:01:32,459 --> 04:01:34,861
PROMOTE CLUSTERING ON THE

4628
04:01:34,861 --> 04:01:37,664
SURFACE OF MATURE VLP,

4629
04:01:37,664 --> 04:01:40,734
SUBSEQUENTLY LEADING TO

4630
04:01:40,734 --> 04:01:43,770
EFFICIENT B CELL STIMULATION BY

4631
04:01:43,770 --> 04:01:46,740
CROSS-LINKING, AND TO EXPRESS

4632
04:01:46,740 --> 04:01:50,143
THE VIRUS PROTEASE WE DECIDED TO

4633
04:01:50,143 --> 04:01:50,844
MIMIC PHYSIOLOGIC MECHANISM

4634
04:01:50,844 --> 04:01:59,152
DURING THE COURSE OF HIV

4635
04:01:59,152 --> 04:02:00,120
INFECTION, TRANSCRIBED AS GAG

4636
04:02:00,120 --> 04:02:01,121
TRANSCRIPT FOLLOWED BY

4637
04:02:01,121 --> 04:02:03,423
TRANSLATION IN A DIFFERENT

4638
04:02:03,423 --> 04:02:12,932
READING FRAME BY RIBOSOMAL SHIFT

4639
04:02:12,932 --> 04:02:13,700
MECHANISM, DESIGNED TO

4640
04:02:13,700 --> 04:02:17,771
IDENTIFIER THE MOST SUITABLE ONE

4641
04:02:17,771 --> 04:02:24,744
TO INDUCE APPROPRIATELY DOSED,

4642
04:02:24,744 --> 04:02:26,079
TIMELY EXPRESSION, FORMATION OF

4643
04:02:26,079 --> 04:02:27,247
MATURE VLPs.

4644
04:02:27,247 --> 04:02:31,551
THREE OF FIVE HAD THE FRAME

4645
04:02:31,551 --> 04:02:33,953
SHIFT SEQUENCES, AND THEY WERE

4646
04:02:33,953 --> 04:02:36,156
OPTIMIZES, CONSTRUCT NUMBER 4

4647
04:02:36,156 --> 04:02:37,690
WAS THE FULL LENGTH CONSTRUCT

4648
04:02:37,690 --> 04:02:39,759
DIVIDE OF THE FRAME SHIFT

4649
04:02:39,759 --> 04:02:43,363
SEQUENCES, FIFTH HAD SOME

4650
04:02:43,363 --> 04:02:45,131
ADDITIONAL TWO-RUN CASE IN PAUSE

4651
04:02:45,131 --> 04:02:45,331
GENE.

4652
04:02:45,331 --> 04:02:53,006
WHEN WE LOOKED AT FOR EFFICIENCY

4653
04:02:53,006 --> 04:02:57,177
BY MEASURING P16 CAPTURABLE B

4654
04:02:57,177 --> 04:02:58,378
2711, HIGHEST LEVEL WITH

4655
04:02:58,378 --> 04:03:01,214
CONSTRUCT NUMBER 4, AGAIN A

4656
04:03:01,214 --> 04:03:04,851
FRAME SHIFT SEQUENCE 3, FULL

4657
04:03:04,851 --> 04:03:07,020
LENGTH GAG POL GENE.

4658
04:03:07,020 --> 04:03:10,423
WE SELECTED THIS CONSTRUCT FOR

4659
04:03:10,423 --> 04:03:12,592
OUR FURTHER STUDIES, NEXT

4660
04:03:12,592 --> 04:03:14,027
DEFINED OPTIMAL AMOUNT FOR

4661
04:03:14,027 --> 04:03:19,466
PRODUCTION OF THE MATURE VERSION

4662
04:03:19,466 --> 04:03:23,236
OF THE VLPs, AND WE

4663
04:03:23,236 --> 04:03:23,970
CO-TRANSFECTED THE ENVELOPE

4664
04:03:23,970 --> 04:03:25,939
EXPRESSING CELL LINE WITH FIXED

4665
04:03:25,939 --> 04:03:30,977
AMOUNT OF GAG mRNA AND

4666
04:03:30,977 --> 04:03:35,515
DECREASING AMOUNT OF GAGPOL,

4667
04:03:35,515 --> 04:03:38,017
EFFICIENCY OF VLP PRODUCTION

4668
04:03:38,017 --> 04:03:39,285
SHOWED DISTRIBUTION, PEAK

4669
04:03:39,285 --> 04:03:43,590
VALUES, THIS PARTICULAR DATA

4670
04:03:43,590 --> 04:03:45,458
SUGGESTED CO-TRANSFECTION OF

4671
04:03:45,458 --> 04:03:48,394
CELL LINES WITH GAG mRNA AND

4672
04:03:48,394 --> 04:03:50,430
GAGPOL N RNA DEVOID OF FRAME

4673
04:03:50,430 --> 04:03:54,634
SHIFT SEQUENCES AT RATIO OF 1 IN

4674
04:03:54,634 --> 04:03:55,735
210 PROVIDE OPTIMAL CONDITION

4675
04:03:55,735 --> 04:03:58,671
FOR PRODUCTION OF MATURE

4676
04:03:58,671 --> 04:03:59,672
VIRUS-LIKE PARTICLES.

4677
04:03:59,672 --> 04:04:02,108
SO, WE WENT AHEAD WITH THE IN

4678
04:04:02,108 --> 04:04:07,247
VITRO EXPRESSION OF IMMATURE AND

4679
04:04:07,247 --> 04:04:08,882
MATURE VLPs FOR

4680
04:04:08,882 --> 04:04:10,083
CHARACTERIZATION, AND THE

4681
04:04:10,083 --> 04:04:12,719
STRUCTURAL CHARACTERISTICS OF

4682
04:04:12,719 --> 04:04:17,390
IMMATURE AND MATURE HIV VLPs

4683
04:04:17,390 --> 04:04:21,361
REVEALED BY NEGATIVE STAINING

4684
04:04:21,361 --> 04:04:22,562
ELECTRON MICROSCOPY, SHOWING

4685
04:04:22,562 --> 04:04:26,032
ARRAY OF ENVELOPE SPIKES ON

4686
04:04:26,032 --> 04:04:28,468
SURFACE, IN DEPTH STRUCTURE

4687
04:04:28,468 --> 04:04:29,802
CHARACTERIZATION OF IMMATURE

4688
04:04:29,802 --> 04:04:33,840
VLPs WAS PERFORMED BY CryoEM

4689
04:04:33,840 --> 04:04:37,777
ANALYSIS, AND THE PARTICLES

4690
04:04:37,777 --> 04:04:40,947
SHOWED A WELL-ORGANIZED GAG

4691
04:04:40,947 --> 04:04:43,149
LAYER, DECORATED WITH THE

4692
04:04:43,149 --> 04:04:44,584
TRIMERS, HERE I'M SHOWING VIDEO

4693
04:04:44,584 --> 04:04:51,891
OF THE TOMOGRAPHIC SLICES.

4694
04:04:51,891 --> 04:04:53,960
INTERESTINGLY FOR MATURE VLPs

4695
04:04:53,960 --> 04:04:55,728
CryoEM IMAGES SHOWED NO

4696
04:04:55,728 --> 04:05:00,333
VISIBLE GAG LAYER, AND WE

4697
04:05:00,333 --> 04:05:01,668
COULDN'T SEE THE CONICAL CODE

4698
04:05:01,668 --> 04:05:04,270
WHICH MAY SUGGEST THE PROCESS IS

4699
04:05:04,270 --> 04:05:05,371
INEFFICIENT, ALTHOUGH SOME

4700
04:05:05,371 --> 04:05:08,641
EVIDENCE OF ENVELOPE CLUSTERING

4701
04:05:08,641 --> 04:05:10,076
WAS OBSERVED.

4702
04:05:10,076 --> 04:05:13,346
SO NEXT TO ASSESS POTENTIAL OF

4703
04:05:13,346 --> 04:05:16,716
ADDITION OF THIS GAGPOL TO

4704
04:05:16,716 --> 04:05:19,953
VACCINE PLATFORM, WE IMMUNIZED

4705
04:05:19,953 --> 04:05:21,354
MICE WITH THE mRNA

4706
04:05:21,354 --> 04:05:22,789
FORMULATION, HAVING ENVELOPE

4707
04:05:22,789 --> 04:05:25,792
ALONE OR IN COMBINATION WITH SIV

4708
04:05:25,792 --> 04:05:30,763
GAG IN THE PRESENCE AND ABSENCE

4709
04:05:30,763 --> 04:05:36,002
OF GAGPOL IN DIFFERENT RATIOS,

4710
04:05:36,002 --> 04:05:46,279
IMMUNIZED WITH .

4711
04:05:47,513 --> 04:05:52,185
AS WE CAN SEE THE TRIMER BINDING

4712
04:05:52,185 --> 04:05:53,820
ANTIBODIES AGAINST AUTOLOGOUS

4713
04:05:53,820 --> 04:05:58,091
ENVELOPE WERE SIGNIFICANTLY AND

4714
04:05:58,091 --> 04:06:02,228
CONSISTENTLY HIGHER FOR THOSE

4715
04:06:02,228 --> 04:06:03,162
ENVs THAT RECEIVED VLP-FORMING

4716
04:06:03,162 --> 04:06:05,298
VACCINE AS COMPARED TO THE ONE

4717
04:06:05,298 --> 04:06:08,167
THAT RECEIVED THE SUBUNIT BASE

4718
04:06:08,167 --> 04:06:10,470
VACCINE, WHICH BASICALLY

4719
04:06:10,470 --> 04:06:12,405
SUGGESTS SUPERIORITY OF THE

4720
04:06:12,405 --> 04:06:14,040
PARTICLE OF VACCINES OVER

4721
04:06:14,040 --> 04:06:16,576
SUBUNIT VACCINES, AND ALSO WE

4722
04:06:16,576 --> 04:06:19,545
OBSERVED THAT ALL THOSE ANIMALS

4723
04:06:19,545 --> 04:06:21,547
THAT RECEIVED PROTEASE

4724
04:06:21,547 --> 04:06:23,483
SUPPLEMENTED VACCINE ELICITED

4725
04:06:23,483 --> 04:06:27,954
SUBSTANTIALLY HIGHER TITERS OF

4726
04:06:27,954 --> 04:06:31,958
ANTIBODIES AS COMPARED TO ENV 2,

4727
04:06:31,958 --> 04:06:34,327
GAG ALONE, DEMONSTRATING THE

4728
04:06:34,327 --> 04:06:35,628
IMPROVED IMMUNOGENICITY OF

4729
04:06:35,628 --> 04:06:37,263
PROTEASE SUPPLEMENTATION TO THE

4730
04:06:37,263 --> 04:06:38,831
VACCINE PLATFORM.

4731
04:06:38,831 --> 04:06:40,867
SO NEXT WE CHECKED

4732
04:06:40,867 --> 04:06:46,172
NEUTRALIZATION ACTIVITY IN THE

4733
04:06:46,172 --> 04:06:51,310
SERUM ANTIBODIES AGAINST

4734
04:06:51,310 --> 04:06:52,345
DEGLYCOSYLATED VERSION OF

4735
04:06:52,345 --> 04:06:52,845
PSEUDOVIRUSES.

4736
04:06:52,845 --> 04:06:57,183
ALL OF THOSE ANIMALS THAT

4737
04:06:57,183 --> 04:06:59,085
RECEIVED PROTEASE SUPPLEMENTED

4738
04:06:59,085 --> 04:07:00,720
REGIMENS ELICITED HIGHER

4739
04:07:00,720 --> 04:07:03,856
NEUTRALIZING ANTIBODIES AS

4740
04:07:03,856 --> 04:07:10,863
COMPARED TO ENV 2, WITH ONE

4741
04:07:10,863 --> 04:07:13,599
HAVING THE LOWEST TITERS.

4742
04:07:13,599 --> 04:07:16,703
NEXT WE AGAIN TESTED THE MOUSE

4743
04:07:16,703 --> 04:07:19,338
SERA FOR ABILITY TO NEUTRALIZE

4744
04:07:19,338 --> 04:07:25,044
THE PARTIALLY CLOSED FORM, A

4745
04:07:25,044 --> 04:07:26,646
COMPETITIVELY MORE CHALLENGING

4746
04:07:26,646 --> 04:07:26,879
TARGET.

4747
04:07:26,879 --> 04:07:28,481
ALTHOUGH WE OBSERVED LIKE

4748
04:07:28,481 --> 04:07:30,717
MARKEDLY LOWER TITERS FOR ALL OF

4749
04:07:30,717 --> 04:07:34,954
THE TIME POINTS, EXCEPT FOR ARM

4750
04:07:34,954 --> 04:07:41,861
3 WHICH RECEIVED THE RATIO OF 1

4751
04:07:41,861 --> 04:07:44,497
IN TWO FIVE SUGGESTING THE

4752
04:07:44,497 --> 04:07:50,002
PLATFORM WHEN SUPPLEMENTED WITH

4753
04:07:50,002 --> 04:07:52,071
PROTEASE mRNA IMPROVED

4754
04:07:52,071 --> 04:07:58,978
EFFICACY OF VACCINE BY INDUCING

4755
04:07:58,978 --> 04:08:04,984
NEUTRALIZING ANTIBODIES. 

4756
04:08:04,984 --> 04:08:06,853
THE INCLUSION FOR PROCESSING GAG

4757
04:08:06,853 --> 04:08:11,791
RESULTED IN MATURE FORM OF

4758
04:08:11,791 --> 04:08:12,525
VLPs ELICITING HIGHER TITERS

4759
04:08:12,525 --> 04:08:15,394
AS COMPARED TO VACCINE THAT

4760
04:08:15,394 --> 04:08:17,930
PRODUCES IMMATURE VLPs,ODOD AND

4761
04:08:17,930 --> 04:08:20,633
THEREFORE IS ONE OF THE IMPROVED

4762
04:08:20,633 --> 04:08:23,336
AND PROMISINGS PLATFORMS FOR HIV

4763
04:08:23,336 --> 04:08:25,705
VACCINE DEVELOPMENT.

4764
04:08:25,705 --> 04:08:27,740
HERE ARE MY ACKNOWLEDGMENTS, MY

4765
04:08:27,740 --> 04:08:31,878
ACKNOWLEDGMENT TO MY LAB MEMBERS

4766
04:08:31,878 --> 04:08:36,082
AND SPECIALISTS, MY POSTDOC

4767
04:08:36,082 --> 04:08:39,652
MENTOR FOR HIS SCHOLARLY INPUT

4768
04:08:39,652 --> 04:08:44,190
AND GUIDANCE, PROVIDING mRNA

4769
04:08:44,190 --> 04:08:48,361
FORMULATION, SPECIAL THANKS FOR

4770
04:08:48,361 --> 04:08:49,395
COLLABORATION, AND WE THANK

4771
04:08:49,395 --> 04:08:52,231
UNIVERSITY OF WASHINGTON FOR

4772
04:08:52,231 --> 04:08:53,566
HELPING US WITH THE CryoEM

4773
04:08:53,566 --> 04:08:55,768
ANALYSIS AND SENDING BEAUTIFUL

4774
04:08:55,768 --> 04:08:56,702
IMAGES OF THE VIRUS-LIKE

4775
04:08:56,702 --> 04:08:57,503
PARTICLES.

4776
04:08:57,503 --> 04:09:00,439
I WOULD ALSO LIKE TO ACKNOWLEDGE

4777
04:09:00,439 --> 04:09:01,808
GATES FOUNDATION, NIAID, OFFICE

4778
04:09:01,808 --> 04:09:03,376
OF AIDS RESEARCH FOR THE

4779
04:09:03,376 --> 04:09:04,076
FINANCIAL ASSISTANCE.

4780
04:09:04,076 --> 04:09:05,211
THANK YOU ALL FOR YOUR KIND

4781
04:09:05,211 --> 04:09:08,247
ATTENTION AND I'LL BE HAPPY TO

4782
04:09:08,247 --> 04:09:08,781
TAKE QUESTIONS IF ANY.

4783
04:09:08,781 --> 04:09:11,284
[APPLAUSE]

4784
04:09:11,284 --> 04:09:16,322
 

4785
04:09:16,322 --> 04:09:20,593
>> DID YOU LOOK AT GAG

4786
04:09:20,593 --> 04:09:24,764
PROCESSING FOR YOUR GAGPOL

4787
04:09:24,764 --> 04:09:24,964
VLPs?

4788
04:09:24,964 --> 04:09:26,966
>> YEAH, WE DID BLOOD
ANALYSIS

4789
04:09:26,966 --> 04:09:29,035
TO BASICALLY LOOK AT GAG

4790
04:09:29,035 --> 04:09:30,002
PROCESSING FOR DIFFERENT AMOUNTS

4791
04:09:30,002 --> 04:09:31,971
BECAUSE OF THE TIME CONSTRAINTS,

4792
04:09:31,971 --> 04:09:33,840
I DIDN'T SHOW THE ENTIRE DATA

4793
04:09:33,840 --> 04:09:38,110
BUT WE DID TRY DIFFERENT AMOUNTS

4794
04:09:38,110 --> 04:09:39,545
OF THE mRNA TO EVALUATE

4795
04:09:39,545 --> 04:09:40,513
OPTIMALITY CONDITION.

4796
04:09:40,513 --> 04:09:42,281
>> WAS THERE EFFICIENT GAG

4797
04:09:42,281 --> 04:09:42,582
PROCESSING?

4798
04:09:42,582 --> 04:09:44,217
BECAUSE THE PARTICLES DID NOT

4799
04:09:44,217 --> 04:09:46,219
LOOK MATURE IN TERMS OF LACKING

4800
04:09:46,219 --> 04:09:46,919
CONICAL CORE.

4801
04:09:46,919 --> 04:09:47,386
>> YEAH, EXACTLY.

4802
04:09:47,386 --> 04:09:50,022
THAT'S ONE OF THE THINGS WE HAVE

4803
04:09:50,022 --> 04:09:51,991
BEEN WORKING ON BECAUSE

4804
04:09:51,991 --> 04:09:53,326
ENHANCING THE AMOUNT OF GAGPOL

4805
04:09:53,326 --> 04:09:57,463
WAS INDUCING SOME AMOUNT OF

4806
04:09:57,463 --> 04:09:59,232
CELLULAR CYTOTOXICITY, THAT'S

4807
04:09:59,232 --> 04:10:02,068
WHY WE CHOSE A SAFER RATIO BUT,

4808
04:10:02,068 --> 04:10:04,136
YES, WE HAVE TO FURTHER OPTIMIZE

4809
04:10:04,136 --> 04:10:07,173
THE CONDITIONS TO GET THE FULLY

4810
04:10:07,173 --> 04:10:08,107
MATURE FORM.

4811
04:10:08,107 --> 04:10:08,541
THANK YOU.

4812
04:10:08,541 --> 04:10:12,411
>> THIS IS A FOLLOW-UP TO
ERIC'S

4813
04:10:12,411 --> 04:10:16,782
QUESTION, EDDIE ARNOLD, RUTGERS

4814
04:10:16,782 --> 04:10:17,750
UNIVERSITY.

4815
04:10:17,750 --> 04:10:21,888
QUESTION IS, HOW DO YOU USE THE

4816
04:10:21,888 --> 04:10:24,090
PROTEASE OF GAGPOL ADDING TO THE

4817
04:10:24,090 --> 04:10:30,630
EQUATION, DO YOU HAVE ANY

4818
04:10:30,630 --> 04:10:32,498
INFORMATION ABOUT WHAT REVERSE

4819
04:10:32,498 --> 04:10:39,605
TRANSCRIPT TASTE MIGHT BE DOING

4820
04:10:39,605 --> 04:10:41,240
BECAUSE PROTEASE IS A SMALL PART

4821
04:10:41,240 --> 04:10:42,008
OF POL.

4822
04:10:42,008 --> 04:10:45,645
>> THAT IS TRUE BUT LIKE
WE TRY

4823
04:10:45,645 --> 04:10:49,415
TO TRUNCATE THE GAGPOL AND DON'T

4824
04:10:49,415 --> 04:10:51,217
SEE LIKE MUCH DIFFERENCE BUT WE

4825
04:10:51,217 --> 04:11:00,059
WE TRY TO KEEP IT AS MINIMAL AS

4826
04:11:00,059 --> 04:11:00,426
POSSIBLE.

4827
04:11:00,426 --> 04:11:01,460
>> JUST WONDERING IF
YOU'RE

4828
04:11:01,460 --> 04:11:02,428
LOOKING INTO THAT ASPECT.

4829
04:11:02,428 --> 04:11:03,062
THANK YOU.

4830
04:11:03,062 --> 04:11:04,597
GREAT TALK.

4831
04:11:04,597 --> 04:11:06,499
>> ONE MORE QUESTION.

4832
04:11:06,499 --> 04:11:08,200
>> QUICK QUESTION ABOUT
THE

4833
04:11:08,200 --> 04:11:14,140
mRNA FOR GAG AND ENV. 

4834
04:11:14,140 --> 04:11:19,178
COVID mRNA VACCINE HAS A RNA

4835
04:11:19,178 --> 04:11:21,314
MODIFICATION TO ENHANCE PROTEIN,

4836
04:11:21,314 --> 04:11:24,784
ALSO AVOID INNATE IMMUNE

4837
04:11:24,784 --> 04:11:25,584
RESPONSE, WONDERED DO YOU HAVE

4838
04:11:25,584 --> 04:11:26,185
MODIFICATION.

4839
04:11:26,185 --> 04:11:28,688
>> THIS IS A MODIFIED
VERSION

4840
04:11:28,688 --> 04:11:34,393
OF mRNA BY MODERNA,

4841
04:11:34,393 --> 04:11:34,727
EXPRESSING --

4842
04:11:34,727 --> 04:11:36,562
>> BOTH GAG AND --

4843
04:11:36,562 --> 04:11:37,463
>> YEAH.

4844
04:11:37,463 --> 04:11:38,230
>> THANK YOU.

4845
04:11:38,230 --> 04:11:40,666
>> IF THERE'S NO OTHER
QUESTIONS

4846
04:11:40,666 --> 04:11:42,668
WE'LL GIVE MAMTA AND THE OTHER

4847
04:11:42,668 --> 04:11:44,670
SPEAKERS A BIG ROUND OF

4848
04:11:44,670 --> 04:11:44,937
APPLAUSE.

4849
04:11:44,937 --> 04:11:45,338
THANK YOU.

4850
04:11:45,338 --> 04:11:47,640
[APPLAUSE]

4851
04:11:47,640 --> 04:11:56,082
 

4852
04:11:56,082 --> 04:12:04,156
 

4853
04:12:04,156 --> 04:12:05,391
>> THANK YOU, EVERYONE,
FOR

4854
04:12:05,391 --> 04:12:05,758
COMING.

4855
04:12:05,758 --> 04:12:07,326
THIS HAS BEEN, AS ALWAYS, A

4856
04:12:07,326 --> 04:12:08,227
GREAT MEETING.

4857
04:12:08,227 --> 04:12:10,396
I ENJOY THIS EVERY YEAR.

4858
04:12:10,396 --> 04:12:14,667
AND I REALLY APPRECIATE ALL YOUR

4859
04:12:14,667 --> 04:12:15,534
PARTICIPATION, ESPECIALLY THE

4860
04:12:15,534 --> 04:12:20,506
POSTERS, HARD TO GET  GET YOU AY

4861
04:12:20,506 --> 04:12:21,607
FROM THE POSTERS, ALWAYS A GOOD

4862
04:12:21,607 --> 04:12:22,475
SIGN.

4863
04:12:22,475 --> 04:12:24,744
I DON'T HAVE ANYTHING TO SAY

4864
04:12:24,744 --> 04:12:27,913
OTHER THAN SEE YOU NEXT YEAR,

4865
04:12:27,913 --> 04:12:29,448
JUNE 23, 2025.

4866
04:12:29,448 --> 04:12:31,217
THIS MEETING IS ALWAYS THE LAST

4867
04:12:31,217 --> 04:12:32,651
MONDAY AND TUESDAY OF THE MONTH,

4868
04:12:32,651 --> 04:12:34,086
IF YOU CAN'T REMEMBER THE DATES.

4869
04:12:34,086 --> 04:12:37,923
AND I HOPE I SEE Y'ALL OUT HERE

4870
04:12:37,923 --> 04:12:39,425
AGAIN, ESPECIALLY SEE IF KEY CAN

4871
04:12:39,425 --> 04:12:42,328
MAKE A RECORD NUMBER OF POSTERS

4872
04:12:42,328 --> 04:12:43,295
AGAIN NEXT YEAR.

4873
04:12:43,295 --> 04:12:45,564
AND GET OUT THERE AND DO SOME

4874
04:12:45,564 --> 04:12:45,998
GOOD SCIENCE.

4875
04:12:45,998 --> 04:12:48,768
[APPLAUSE]

4876
04:12:48,768 --> 04:12:51,704
[END OF PROGRAM]

4877
04:12:51,704 --> 04:12:57,743
 

4878
04:12:57,743 --> 04:13:07,787