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GOOD AFTERNOON.

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I'M STEVE HUGHES AND I WANT TO 

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WELCOME ALL OF YOU TO THE DAVE 

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DERSE MEMORIAL LECTURE.

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DAVE HAS NOW BEEN GONE FOR 15 

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YEARS, AND THAT MEANS, 

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UNFORTUNATELY, THAT MANY OF YOU 

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DID NOT HAVE A CHANCE TO GET TO 

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KNOW DAVE PERSONALLY.

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SO THAT BRINGS ME TO THE 

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QUESTION OF WHO DAVE DERSE WAS 

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AND WHY WE WANT TO BOTH REMEMBER

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AND HONOR HIM.

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DAVE WAS A SCHOLAR AND HE LOVED 

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THE INTELLECTUAL CHALLENGES THAT

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SCIENCE PROVIDES.

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FOR THAT REASON, IT IS ENTIRELY 

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FITTING THAT WE HONOR HIS MEMORY

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IN A FORM THAT DAVE WOULD 

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PARTICULARLY HAVE LIKE, A 

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LECTURE FROM CAROL CARTER.

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THIS IS DAVE AND HIS COLLEAGUES 

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IN, OH, MAYBE 20 YEARS AGO.

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DAVE WAS ALSO NOT ONLY A 

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VALUABLE MENTOR, WAS DEEPLY 

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DEDICATED TO PEOPLE WHO WORK 

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WITH HIM, HE WAS A COLLEAGUE AND

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THOSE OF US WHO WORKED WITH HIM 

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GREATLY MISS HIM.

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HERE HE'S SHOWN WITH ANOTHER 

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MEMBER OF THE DRP IN THE 

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AUDIENCE TODAY, WHO I'M SURE 

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MISSING HIM JUST AS MUCH AS THE 

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REST OF US, BUT MOST OF ALL, 

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DAVE WAS A FRIEND.

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WE SHOULD REMEMBER DAVE IN THE 

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LARGER SENSE.

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WE WANT TO REMEMBER HIM, OF 

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COURSE, AS A SCIENTIST, BUT WE 

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WANT TO REMEMBER HIM NOT ONLY AS

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A SCIENTIST, BUT ALSO AS A 

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PERSON.

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DAVE WAS REMARKABLY KIND AND 

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AGAIN TELL.

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HE HAD A -- GENTLE.

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HE HAD A MARVELOUS CLARITY OF 

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SPIRIT.

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I CAN SAY HONESTLY THAT I NEVER 

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HEARD HIM SAY A HARSH WORD ABOUT

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ANYONE.

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BECAUSE WE SHARED A LOT OF THE 

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OUTDOORS AND FISHING, I HAD THE 

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PRIVILEGE OF SPENDING TIME WITH 

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HIM AWAY FROM THE LAB.

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HERE'S DAVE WITH A REALLY NICE 

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STRIPER CAUGHT OUT IN CHESAPEAKE

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BAY, AGAIN ABOUT 20 YEARS AGO.

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IT REALLY WASN'T THE SIZE OF THE

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FISH THAT MATTERS.

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HERE'S DAVE WITH A LITTLE BROWN 

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HE CAUGHT ON COTTON CREEK AND 

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HIS SMILE WAS EVERY BIT AS BIG 

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AS IT WAS WHEN HE HAD THE BIG 

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STR

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STRIPER.

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AND THOSE OF US WHO KNEW DAVE 

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WELL HAVE MANY FOND MEMORIES OF 

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HIM.

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I CAN GO BACK TO CHESAPEAKE OR 

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TO JOHN CREEK AND SOME OF THE 

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OTHER PLACES WE FISH AND BE 

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REMINDED OF WHAT A WONDERFUL 

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FRIEND AND COLLEAGUE HE WAS.

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AND IT'S PARTICULARLY IMPORTANT,

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I THINK, THAT WE SHOULD HONOR 

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HIS KINDNESS AND HIS GENTLENESS 

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AS WE APPROACH AN ELECTION THAT 

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IS REMARKABLY BITTER.

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IT'S FILLED WITH HARSH WORDS AND

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THE THREAT OF EVEN HARSHER 

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DEEDS.

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SO I WOULD HOPE THAT DAVE IS 

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WITH US TODAY IN SPIRIT AND HIS 

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GENTLE APPROACH TO LIFE CAN HELP

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US NAVIGATE THESE TROUBLED 

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TIMES.

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WELCOME TO THE LECTURE THANK 

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YOU.

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[ APPLAUSE ]

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>> THANK YOU VERY MUCH FOR
THAT 

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STEVE.

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WE CERTAINLY DO MISS DAVE VERY 

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MUCH.

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I'D LIKE TO JOIN STEVE AND ALL 

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THE FACULTY OF THE HIV DRP IN 

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WELCOMING YOU TO THE 13th ANNUAL

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DAVID DERSE MEMORIAL LECTURE AND

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AWARD.

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AS STEVE ALLUDED TO, MANY IN 

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THIS AUDIENCE DID NOT KNOW DAVE 

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PERSONALLY.

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TO LEARN MORE ABOUT HIS LIFE AND

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HIS CAREER, I WOULD DIRECT YOU 

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TO THIS COMMENTARY WRITTEN BY 

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ONE OF HIS TRAINEES, MAUREEN 

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SHUH, PUBLISHED IN 2009, SHORTLY

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AFTER HIS UNTIMELY PASSING.

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I'D LIKE TO SHARE ONE EXCERPT 

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FROM THIS COMMENTARY.

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DAVE WAS VERY PASSIONATE ABOUT 

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SCIENCE AND HIS ENTHUSIASM FOR 

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WORK WAS CONTAGIOUS TO MEMBERS 

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OF THE LABORATORY.

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DAVE MADE SEVERAL IMPORTANT 

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CONTRIBUTIONS TO THE FIELD, BUT 

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HE NEVER SOUGHT ACCOLADES FOR 

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HIMSELF.

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DAVE REMAINS THE MOST GENUINE 

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AND KIND INDIVIDUAL I KNOW.

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I THINK ALL OF US WHO KNEW DAVE 

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FELT THIS WAY ABOUT HIM.

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SO I RECENTLY HAD THE 

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OPPORTUNITY TO COMB THROUGH THE 

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COLD SPRING HARBOR PHOTO 

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ARCHIVES AND FOUND A FEW OLD 

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PHOTOS OF DAVE THAT I 

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PARTICULARLY LIKED.

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ONE HERE ON THE LEFT TAKEN IN 

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1992, AND TWO IN 1999, INCLUDING

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THE ONE ON THE RIGHT OF DAVE AND

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KATE, WHO LATER BECAME KATE 

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DERSE AND REMAINS ONE OF THE 

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MAJOR SPONSORS OF THIS EVENT.

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DAVE CONTRIBUTED IN SUBSTANTIAL 

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WAYS IN MANY ASPECTS OF 

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VIROLOGY.

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EARLY IN HIS CAREER, I STUDIED 

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HSV POLYMERASE BEFORE SWITCHING 

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TO RETROVIRUSES, GENE EXPRESSION

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AND RNA SURPRISING.

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HE DEVELOPED A NUMBER OF 

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RETROVIRAL VECTORS AND CLONES 

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STILL USED TODAY.

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HE STUDIED RETROVIRAL ASSEMBLY 

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AND TRANSMISSION AND RESTRICTION

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OF RETROVIRAL INFECTIVITY BY 

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APOBEC.

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THE DERSE MEMORIAL LECTURE AND 

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AWARD WERE ESTABLISHED IN 2012 

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AND FEATURED A NUMBER OF 

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PROMINENT SCIENTISTS, INCLUDING 

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PAT GREEN, JOE GIAM, WALTER 

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MOTHES, QUENTIN SATTEN TAU, 

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CHARLES, BEATRICE, WE WENT 

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VIRTUAL DURING COVID IN 2020 AND

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2021.

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AND THEN WE RESUMED IN-PERSON 

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MEETINGS IN 2022 WITH DARIA AND 

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LAST YEAR WITH LOU MANSKY.

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SO THAT BRINGS US TO THIS YEAR'S

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LECTURE AND AWARDEE, CAROL 

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CARTER.

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CAROL RECEIVED HER BACHELORS IN 

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DEGREE IN BIOLOGY AND CHEMISTRY 

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FROM 1967 FROM CITY COLLEGE OF 

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NEW YORK.

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WENT ON TO GET HER PhD IN 

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MICROBIOLOGY FROM YALE.

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DID A TWO-YEAR POST DOC AT ROCHE

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AND WAS RESKRUTED AS ASSISTANT 

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PROFESSOR TO STONY BOOK 

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UNIVERSITY WHERE SHE'S REMAINED 

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EVER SINCE, CLIMBING THROUGH THE

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RANKS, BECOMING FULL PROFESSOR 

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IN 1994 AND IN 2022, THE STATE 

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UNIVERSITY OF NEW YORK 

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DISTINGUISHED PROFESSOR AT STONY

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BROOK.

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CAROL HAS RECEIVED A NUMBER OF 

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HONORS AND RECOGNITION AND IS 

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WELL-KNOWN FOR HER SERVICE TO 

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THE COMMUNITY.

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IN 2013, SHE RECEIVED THE STONY 

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BROOK UNIVERSITY PRESIDENTIAL 

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AWARD FOR PROMOTING DIVERSITY 

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AND ACADEMIC EXCELLENCE.

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TWICE, SHE LEAFED THE STONY 

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BROOK EXCEPTIONAL SERVICE TO 

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UNDERGRADUATE EDUCATION AWARD.

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HAS SERVED ON NUMEROUS EDITORIAL

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BOARDS AND STUDY SECTIONS.

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IN 2022, WAS ELECTED TO THE 

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AMERICAN ACADEMY OF MICROBIOLOGY

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AND EARLIER THIS YEAR TO THE 

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NATIONAL ACADEMY OF SCIENCES.

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CONGRATULATIONS TO CAROL FOR 

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THAT WELL-DESERVED HONOR.

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[ APPLAUSE ]

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CAROL'S REACH HAS SPANNED A WIDE

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SWATH OF RETROVIROLOGY, 

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ACTIVATION OF THE HIV-1 PROCEED 

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TEEATION, THE STRUCTURE OF 

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RETROVIRAL PROTEINS, THE ROLE OF

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ESCRT MACHINERY, THE ROLE OF 

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CALCIUM AND IS FOR TOINNOSITIDE 

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SIGNALING.

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SO CAROL IS NOT ONLY A 

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WELL-KNOWN AND DISTINGUISH 

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SCIENTIST, BUT SHE'S ALSO A 

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CARING MENTOR AND ROLE MODEL FOR

211
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YOUNGER PEOPLE IN THE COMMUNITY.

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I THINK THIS IS CAPTURED TO SOME

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EXTENT IN THESE PHOTOS TAKEN 

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WITH CAROL WITH MEMBERS OF MY 

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LAB AT COLD SPRING HARBOR BACK 

216
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IN MAY.

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YOU CAN SEE THEY'RE HAVING A 

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GOOD TIME HERE.

219
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[ LAUGHTER ]

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SO WITHOUT ANY FURTHER DELAY, I 

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WOULD LIKE TO WELCOME CAROL TO 

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THE PODIUM FOR HER LECTURE 

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ENTITLED HOST-TARGETED 

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THERAPEUTICS, PRAZOLES TARGET 

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TSG101 UBIQUITIN E2 VARIANT 

226
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MIMICRY, AND TO RECEIVE HER 

227
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DAVID DERSE MEMORIAL LECTURE 

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AWARD.

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[ APPLAUSE ]

230
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IT'S ALL YOUR FAULT.

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[ LAUGHTER ]

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>> THANK YOU.

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READY.

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1, 2, 3.

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PERFECT.

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A COUPLE LIKE THIS.

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1, 2, 3.

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AND 1, 2, 3.

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[ APPLAUSE ]

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>> SO ONCE AGAIN, THANK
YOU, 

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THANK YOU, THANK YOU.

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THIS IS A VERY SPECIAL HONOR FOR

243
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ME BECAUSE I KNOW SO MANY 

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FABULOUS SCIENTISTS WHO ARE HERE

245
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AND DIDN'T ACTUALLY EVER MEET 

246
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DAVID DURING HIS LIFE, BUT 

247
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CERTAINLY ACQUAINTED WITH THE 

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WORK THAT HE DID.

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SO I'M GOING TO START OFF.

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STEVE HUGHES JUST SAID TO ME, HE

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SAID KNOCK THEM DEAD.

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WELL, I'M GOING TO START OFF BY 

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SIMPLIFYING MY TITLE.

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SO HOST-TARGETED THERAPEUTICS, 

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PRAZOLES TARGET EXIT, AND THAT 

256
00:09:46,743 --> 00:09:50,380
IS WHAT WE'RE GOING TO SORT OF 

257
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DO TO TALK TO YOU ABOUT TODAY 

258
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AND ONE OF THE THINGS THAT HAS 

259
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ALWAYS FASCINATED ME.

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SO EVERYBODY WHO IS IN THIS ROOM

261
00:10:01,491 --> 00:10:04,594
OR VIRTUAL IS ACQUAINTED, I 

262
00:10:04,594 --> 00:10:05,595
THINK, FOR THE MOST PART WITH 

263
00:10:05,595 --> 00:10:08,699
HIV, ITS LIFECYCLE, AND THE 

264
00:10:08,699 --> 00:10:11,902
INCREDIBLE AND IMPRESSIVE 

265
00:10:11,902 --> 00:10:13,303
COMPLEXITY THAT THIS VIRUS 

266
00:10:13,303 --> 00:10:14,738
PRESENTS IN DOING WHAT IT 

267
00:10:14,738 --> 00:10:15,939
NATURALLY DOES FOR ITSELF, 

268
00:10:15,939 --> 00:10:18,542
REPLICATE.

269
00:10:18,542 --> 00:10:19,776
AND ONE OF THE THINGS THAT'S 

270
00:10:19,776 --> 00:10:23,313
ALWAYS FASCINATED ME IS THAT 

271
00:10:23,313 --> 00:10:26,583
EVERY STEP OF ITS REPLICATION 

272
00:10:26,583 --> 00:10:30,153
CYCLE IS IMPORTANT, BUT IN 

273
00:10:30,153 --> 00:10:33,190
PARTICULAR, THE LAST STEP, THE 

274
00:10:33,190 --> 00:10:37,494
EGRESS STEP IS NOT ONLY WHAT 

275
00:10:37,494 --> 00:10:39,529
IMPORTANT IN ITS OWN RIGHT, BUT 

276
00:10:39,529 --> 00:10:40,397
IT'S CRITICALLY IMPORTANT 

277
00:10:40,397 --> 00:10:42,032
BECAUSE IT SETS THE VIRUS UP FOR

278
00:10:42,032 --> 00:10:44,067
WHAT IT'S GOT TO DO IN THE NEXT 

279
00:10:44,067 --> 00:10:47,037
STAGE, WHICH IS TO GET INTO 

280
00:10:47,037 --> 00:10:47,738
ANOTHER CELL.

281
00:10:47,738 --> 00:10:51,074
SO I TOOK A QUOTE FROM 

282
00:10:51,074 --> 00:10:55,545
SHAKESPEARE'S MAC BETH, NOTHING 

283
00:10:55,545 --> 00:10:57,147
IN THE VIRUS' LIFECYCLE BECOMES 

284
00:10:57,147 --> 00:10:58,682
IT LIKE THE LEAVING OF IT.

285
00:10:58,682 --> 00:10:59,983
IN OTHER WORDS, IT'S GOT TO DO 

286
00:10:59,983 --> 00:11:03,153
IT RIGHT OR ELSE IT'S DONE.

287
00:11:03,153 --> 00:11:05,922
AND SO WE -- THE FOCUS OF THE 

288
00:11:05,922 --> 00:11:09,259
TALK TODAY IS REALLY GOING TO BE

289
00:11:09,259 --> 00:11:10,293
WHAT'S INVOLVED IN GETTING OUT 

290
00:11:10,293 --> 00:11:12,329
OF AN INFECTED CELL.

291
00:11:12,329 --> 00:11:14,398
IT'S NOT AS EASY AS IT MIGHT 

292
00:11:14,398 --> 00:11:18,935
SEEM TO BE.

293
00:11:18,935 --> 00:11:22,739
SO BACK, I GUESS, A COUPLE OF 

294
00:11:22,739 --> 00:11:27,711
DECADES AGO NOW, THE FIELD WAS 

295
00:11:27,711 --> 00:11:30,680
VERY ANXIOUS TO TRY TO IDENTIFY 

296
00:11:30,680 --> 00:11:33,850
SOME ANTIVIRALS THAT WOULD 

297
00:11:33,850 --> 00:11:36,253
TARGET HIV REPLICATION, AND IT 

298
00:11:36,253 --> 00:11:38,522
WAS SORT OF, IN MY OPINION, WAS 

299
00:11:38,522 --> 00:11:41,324
SORT OF A STOPGAP MEASURE 

300
00:11:41,324 --> 00:11:44,294
BECAUSE, AFTER ALL, WE'RE GOING 

301
00:11:44,294 --> 00:11:45,996
TO HAVE A VACCINE ANY DAY, SO WE

302
00:11:45,996 --> 00:11:47,364
CAN JUST BUSY OURSELVES WITH 

303
00:11:47,364 --> 00:11:49,366
GETTING SOME ANTIVIRALS GOING 

304
00:11:49,366 --> 00:11:50,934
THAT WOULD SORT OF HOLD US UNTIL

305
00:11:50,934 --> 00:11:54,805
THE VACCINE, YOU KNOW, WAS 

306
00:11:54,805 --> 00:11:56,273
AVAILABLE.

307
00:11:56,273 --> 00:12:00,844
AND AT THE TIME, THE FIELD AND 

308
00:12:00,844 --> 00:12:02,279
IN PARTICULAR WITH 

309
00:12:02,279 --> 00:12:04,147
COLLABORATIONS FROM SEVERAL 

310
00:12:04,147 --> 00:12:06,516
COMPANIES, DID WHAT WAS 

311
00:12:06,516 --> 00:12:10,687
SUPPOSABLE STRATEGY, WHICH WAS 

312
00:12:10,687 --> 00:12:13,990
TO TARGET THE GENES THAT ARE 

313
00:12:13,990 --> 00:12:15,525
ENCODED BY THE VIRUS ITSELF AND 

314
00:12:15,525 --> 00:12:18,495
TO TARGET THEM FOR DEVELOPMENT 

315
00:12:18,495 --> 00:12:21,698
OF AGENTS THAT WOULD INTERRUPT 

316
00:12:21,698 --> 00:12:23,366
THEIR FUNCTION.

317
00:12:23,366 --> 00:12:26,870
NOW, AT THE TIME, IF THE IDEA OF

318
00:12:26,870 --> 00:12:29,706
TARGETING NOT JUST THE VIRAL 

319
00:12:29,706 --> 00:12:33,443
ENCODED GENES, BUT ALSO THE HOST

320
00:12:33,443 --> 00:12:34,311
GENES THAT ACTUALLY HELP THE 

321
00:12:34,311 --> 00:12:38,248
VIRUS TO DO WHAT IT DOES, THAT 

322
00:12:38,248 --> 00:12:44,187
IS TO SAY, THE VIRAL GENES THAT 

323
00:12:44,187 --> 00:12:47,257
ENABLE THE VIRUS TO EXPLOIT HOST

324
00:12:47,257 --> 00:12:47,891
MACHINERY, WE WOULD HAVE 

325
00:12:47,891 --> 00:12:50,127
PROBABLY BEEN LAUGHED OUT OF THE

326
00:12:50,127 --> 00:12:51,461
ROOM, AND IT WASN'T UNTIL 

327
00:12:51,461 --> 00:12:53,396
PROBABLY AROUND, I THINK, THE 

328
00:12:53,396 --> 00:12:57,868
MID TO LATE '90s, WHEN IT BECAME

329
00:12:57,868 --> 00:13:01,905
APPARENT THAT THIS PROBLEM OF 

330
00:13:01,905 --> 00:13:03,006
DRUG-RESISTANCE THAT FOLLOWED 

331
00:13:03,006 --> 00:13:05,709
THE TARGETING OF ANY COMPOUND, 

332
00:13:05,709 --> 00:13:08,879
WAS NOT GOING TO GO AWAY.

333
00:13:08,879 --> 00:13:11,848
AND SO WE ENTERTAINED THE IDEA 

334
00:13:11,848 --> 00:13:14,484
THAT MAYBE -- AND WE STILL DON'T

335
00:13:14,484 --> 00:13:16,920
KNOW, BUT MAYBE WE TARGETED HOST

336
00:13:16,920 --> 00:13:19,256
GENE PRODUCTS THAT WERE HELPING 

337
00:13:19,256 --> 00:13:22,759
THE VIRUS, THEY WOULD BE NOT SO 

338
00:13:22,759 --> 00:13:26,296
EASY TO ACCRUE RESISTANCE TO.

339
00:13:26,296 --> 00:13:30,267
SO WE STARTED TO LOOK FOR THEM.

340
00:13:30,267 --> 00:13:31,801
WE WERE VERY FORTUNATE THAT THE 

341
00:13:31,801 --> 00:13:34,104
PERSON OF THE INVESTOR OF THESE 

342
00:13:34,104 --> 00:13:36,740
TWO ASSAY HYBRIDS, WAS AN 

343
00:13:36,740 --> 00:13:37,841
ASSISTANT PROFESSOR DOWN ON THE 

344
00:13:37,841 --> 00:13:40,577
FIRST FLOOR IN MY BUILDING, AND 

345
00:13:40,577 --> 00:13:42,479
WAS VERY ANXIOUS FOR PEOPLE TO 

346
00:13:42,479 --> 00:13:45,282
USE HIS ASSAY, AND SO WHAT 

347
00:13:45,282 --> 00:13:48,018
ESSENTIALLY WE DID WAS TO SEARCH

348
00:13:48,018 --> 00:13:51,621
A B CELL LIBRARY FOR PROTEINS 

349
00:13:51,621 --> 00:13:53,490
THAT WOULD INTERACT WITH THE 

350
00:13:53,490 --> 00:13:55,559
STRUCTURAL PROTEIN ENCODED BY 

351
00:13:55,559 --> 00:13:58,328
HIV, THE PROTEIN THAT'S NOT ONLY

352
00:13:58,328 --> 00:14:02,199
RESPONSIBLE FOR MAKING VIRAL 

353
00:14:02,199 --> 00:14:05,902
PARTICLES, BUT ALSO FORTUNATE, 

354
00:14:05,902 --> 00:14:08,104
TO MAKE SOMETHING THAT AT LEAST 

355
00:14:08,104 --> 00:14:11,575
LOOKS LIKE AN IMMATURE VIRUS 

356
00:14:11,575 --> 00:14:12,909
PARTICLE, WE USED B CELL LIBRARY

357
00:14:12,909 --> 00:14:20,250
TO FISH OUT PARTNERS OF THAT --.

358
00:14:20,250 --> 00:14:23,787
NOW, IN THE INITIAL FORAYS, WE 

359
00:14:23,787 --> 00:14:26,623
PULLED OUT SEVERAL HOST GENE 

360
00:14:26,623 --> 00:14:27,290
PRODUCTS AND THE GRADUATE 

361
00:14:27,290 --> 00:14:33,029
STUDENT WHO WAS WORKING ON THIS 

362
00:14:33,029 --> 00:14:37,000
PROJECT, SHE PULLED OUT ACTUALLY

363
00:14:37,000 --> 00:14:38,501
CYCLOCILLIN, AND AT THE TIME, 

364
00:14:38,501 --> 00:14:41,171
THEY WERE WORKING ON CYCLOCILLIN

365
00:14:41,171 --> 00:14:43,506
AND BETH SAID, I'M NOT GOING 

366
00:14:43,506 --> 00:14:45,275
TO -- YOU'RE NOT GOING TO TALK 

367
00:14:45,275 --> 00:14:47,344
ME INTO DOING MY THESIS ON A 

368
00:14:47,344 --> 00:14:49,212
PROTEIN THAT NOBODY HEARD OF AND

369
00:14:49,212 --> 00:14:50,413
IS NOT IN THE DATABASE.

370
00:14:50,413 --> 00:14:55,518
I'M GOING TO WRITE A THESIS ON 

371
00:14:55,518 --> 00:14:56,286
CYCLOCILLIN.

372
00:14:56,286 --> 00:15:00,557
BUT FORTUNATELY, AS GRADUATE 

373
00:15:00,557 --> 00:15:02,158
STUDENTS GO, WE WERE ABLE TO 

374
00:15:02,158 --> 00:15:03,560
TALK THE NEXT GRADUATE STUDENT 

375
00:15:03,560 --> 00:15:05,762
WHO JOINED THE LAB INTO WORKING 

376
00:15:05,762 --> 00:15:08,131
ON THIS PROTEIN THAT BETH HAD 

377
00:15:08,131 --> 00:15:10,000
PULLED THE SEQUENCE OUT OF THE 

378
00:15:10,000 --> 00:15:11,434
DATABASE, WAS LOOKING LIKE -- IT

379
00:15:11,434 --> 00:15:14,537
WAS -- I'M SORRY, SHE PULLED OUT

380
00:15:14,537 --> 00:15:16,539
OF THE HYBRID ANALYSIS, THAT THE

381
00:15:16,539 --> 00:15:19,609
SEQUENCE WAS NOT IN PROTEIN 

382
00:15:19,609 --> 00:15:20,176
DATABASE.

383
00:15:20,176 --> 00:15:22,612
SO WHAT WE STARTED WAS -- WE 

384
00:15:22,612 --> 00:15:24,180
WANTED TO UNDERSTAND WHETHER HIV

385
00:15:24,180 --> 00:15:26,149
CARED BIT AND IN WHAT WAY MIGHT 

386
00:15:26,149 --> 00:15:31,388
IT BE UTILIZING IT SUCH THAT WE 

387
00:15:31,388 --> 00:15:32,355
SAW INTERACTION WITH GAG.

388
00:15:32,355 --> 00:15:35,625
SO AS WE WERE WORKING ON THIS, 

389
00:15:35,625 --> 00:15:36,960
WE KEPT EVERY MONTH OR SO WHERE 

390
00:15:36,960 --> 00:15:38,628
WE GO TO THE PROTEIN DATABASE 

391
00:15:38,628 --> 00:15:43,266
AND LOOK TO SEE IF THE SEQUENCE 

392
00:15:43,266 --> 00:15:46,770
OF TSG101 HAD IN FACT BEEN 

393
00:15:46,770 --> 00:15:48,872
ENTERED MACE IN THE DATABASE BY 

394
00:15:48,872 --> 00:15:50,874
ANYBODY, AND ONE DAY, IT 

395
00:15:50,874 --> 00:15:51,374
APPEARED.

396
00:15:51,374 --> 00:15:55,679
IT HAD BEEN PLACED THERE BY A 

397
00:15:55,679 --> 00:16:00,784
CANCER GENETICIC STAN COHEN, WHO

398
00:16:00,784 --> 00:16:02,252
WAS AT STANFORD.

399
00:16:02,252 --> 00:16:05,455
WHEN I SAW THE SEQUENCE IN THE 

400
00:16:05,455 --> 00:16:08,358
DATABASE, I CALLED HIM AND HE 

401
00:16:08,358 --> 00:16:11,127
EXPLAINED TO ME HOW HE HAD 

402
00:16:11,127 --> 00:16:13,463
HAPPENED TO COME ACROSS TSG101.

403
00:16:13,463 --> 00:16:16,232
ESSENTIALLY, HE HAD MADE A 

404
00:16:16,232 --> 00:16:19,736
STRAIN OF OLIGO KNEW THEO TIDES.

405
00:16:19,736 --> 00:16:22,572
HE HAD INJECTED THOSE INTO NICE 

406
00:16:22,572 --> 00:16:26,710
AND HE LOOKED TO SEE WHICH MICE 

407
00:16:26,710 --> 00:16:29,579
DEVELOPED TUMORS BECAUSE THE 

408
00:16:29,579 --> 00:16:33,316
OLIGONUCLEOTIDES WERE ACTING AS 

409
00:16:33,316 --> 00:16:33,817
AN ANTIPHASE.

410
00:16:33,817 --> 00:16:35,652
INSURE ENOUGH, HE PULLED OUT, 

411
00:16:35,652 --> 00:16:38,955
THE MOUSE HAD A TUMOR AND THE 

412
00:16:38,955 --> 00:16:42,025
CARBON SEQUENCE WAS TSG101.

413
00:16:42,025 --> 00:16:44,761
SO THIS WAS FASCINATING BECAUSE 

414
00:16:44,761 --> 00:16:47,130
HERE HE WAS KNOCKING OUT A GENE 

415
00:16:47,130 --> 00:16:53,636
THAT CAUSED A CELL TO OVERGROW, 

416
00:16:53,636 --> 00:16:54,204
ESSENTIALLY.

417
00:16:54,204 --> 00:16:56,573
HERE WE WERE WORKING WITH THIS 

418
00:16:56,573 --> 00:16:59,442
SAME GENE PRODUCT AND AT THAT 

419
00:16:59,442 --> 00:17:02,779
POINT, WE KNEW THAT HIV ACTUALLY

420
00:17:02,779 --> 00:17:05,148
NEEDED TSG101 TO REPLICATE.

421
00:17:05,148 --> 00:17:09,386
SO WE WERE SAYING, WELL, MAYBE 

422
00:17:09,386 --> 00:17:12,655
THEN, THIS IS A SITUATION WHERE 

423
00:17:12,655 --> 00:17:14,491
THE VIRUS IS ACTUALLY EXPLOITING

424
00:17:14,491 --> 00:17:20,196
THE PROTEIN IN A WAY THAT IS A 

425
00:17:20,196 --> 00:17:20,430
NICHE.

426
00:17:20,430 --> 00:17:22,866
BY A NICHE, I MEAN SOME WAY IN 

427
00:17:22,866 --> 00:17:24,734
WHICH IT'S EXPLOITING THE 

428
00:17:24,734 --> 00:17:25,735
PROTEIN, IT'S A CELLULAR 

429
00:17:25,735 --> 00:17:27,404
PROTEIN, AN IMPORTANT CELLULAR 

430
00:17:27,404 --> 00:17:28,838
PROTEIN, BUT YET THE VIRUS IS 

431
00:17:28,838 --> 00:17:31,241
USING IT IN SOME WAY THAT WE 

432
00:17:31,241 --> 00:17:34,477
MIGHT THEN BE ABLE TO ULTIMATELY

433
00:17:34,477 --> 00:17:37,414
TARGET IT BECAUSE IT IS ONE THAT

434
00:17:37,414 --> 00:17:39,716
THE VIRUS USES IN A WAY THAT'S 

435
00:17:39,716 --> 00:17:41,284
NOT QUITE IDENTICAL TO THE WAY 

436
00:17:41,284 --> 00:17:43,620
THE HOST IS USING IT.

437
00:17:43,620 --> 00:17:47,223
SO THAT REALLY BECAME THE SORT 

438
00:17:47,223 --> 00:17:49,726
OF DRIVING HYPOTHESIS IN THE LAB

439
00:17:49,726 --> 00:17:51,561
THAT WE THEN FOLLOWED FOR A 

440
00:17:51,561 --> 00:17:54,230
COUPLE OF DECADES.

441
00:17:54,230 --> 00:17:56,065
NOW, JUST ABOUT THE SAME TIME, 

442
00:17:56,065 --> 00:17:58,368
BECAUSE WE ALWAYS LIKED TO DO 

443
00:17:58,368 --> 00:17:59,636
COMPARATIVE VIROLOGY, WE 

444
00:17:59,636 --> 00:18:01,671
ALWAYS -- IF SOMETHING IS GOING 

445
00:18:01,671 --> 00:18:03,907
ON WITH HIV, WE LIKE TO KNOW WAS

446
00:18:03,907 --> 00:18:05,842
IT ALSO HAPPENING WITH THE 

447
00:18:05,842 --> 00:18:06,976
CHICKEN VIRUS, THE MOUSE VIRUS, 

448
00:18:06,976 --> 00:18:11,114
THE MONKEY VIRUS, THE HORSE 

449
00:18:11,114 --> 00:18:16,786
VIRUS, AND WE THEN, AT SOME 

450
00:18:16,786 --> 00:18:20,957
POINT, WE WENT TO THE CULTURE 

451
00:18:20,957 --> 00:18:21,191
MEETING.

452
00:18:21,191 --> 00:18:23,026
I -- COLD SPRING HARBOR MEETING.

453
00:18:23,026 --> 00:18:24,294
I REMEMBER BEING A LITTLE THE 

454
00:18:24,294 --> 00:18:28,531
COLD SPRING HARBOR, ANOTHER 

455
00:18:28,531 --> 00:18:29,566
RETROVIRAL GIST SAID CAROL, THIS

456
00:18:29,566 --> 00:18:31,634
IS REALLY INTERESTING, WHEN I GO

457
00:18:31,634 --> 00:18:36,773
BACK HOME, I'M GOING TO WORK ON 

458
00:18:36,773 --> 00:18:38,808
IT WITH -- WE SAID DON'T BOTHER,

459
00:18:38,808 --> 00:18:39,976
JOHN, WE ALREADY KNOW.

460
00:18:39,976 --> 00:18:41,945
WE DECIDED TO COLLABORATE AND IN

461
00:18:41,945 --> 00:18:42,979
COLLABORATING, WE DID SOMETHING 

462
00:18:42,979 --> 00:18:45,682
THAT WAS VERY, VERY INFORMATIVE 

463
00:18:45,682 --> 00:18:47,617
FOR US.

464
00:18:47,617 --> 00:18:53,223
SO THE AVIAN RETROVIRUS IS A 

465
00:18:53,223 --> 00:18:56,726
RETROVIRUS THAT CALLS ESCRT 

466
00:18:56,726 --> 00:18:57,760
MACHINERY BY A -- I'LL TALK MORE

467
00:18:57,760 --> 00:19:01,397
ABOUT IN A BIT, WHAT'S CALLED 

468
00:19:01,397 --> 00:19:03,766
THE PR MOTIF.

469
00:19:03,766 --> 00:19:07,637
HIV DOES NOT HAVE ONE OF THOSE.

470
00:19:07,637 --> 00:19:11,374
SO IT SEEMED AS THOUGH MAYBE 

471
00:19:11,374 --> 00:19:13,476
EVEN THOUGH THEY BOTH COULD CALL

472
00:19:13,476 --> 00:19:16,246
ESCRT MACHINERY, WERE THEY DOING

473
00:19:16,246 --> 00:19:17,680
IT BY SOME DIFFERENT KIND OF 

474
00:19:17,680 --> 00:19:18,715
MECHANISM OR MIGHT THESE BE 

475
00:19:18,715 --> 00:19:20,383
MECHANISMS THAT ARE GOING TO 

476
00:19:20,383 --> 00:19:20,717
OVERLAP?

477
00:19:20,717 --> 00:19:22,519
THAT WAS A SORT OF A QUESTION 

478
00:19:22,519 --> 00:19:23,953
THAT WE WANTED TO TRY TO 

479
00:19:23,953 --> 00:19:25,788
UNDERSTAND, BUT AT THE SAME 

480
00:19:25,788 --> 00:19:28,391
TIME, AS PEOPLE STUDIED IN MANY 

481
00:19:28,391 --> 00:19:32,028
LABS INVOLVED IN THIS, AS PEOPLE

482
00:19:32,028 --> 00:19:34,731
STUDIED RETROVIRUSES, INCLUDING 

483
00:19:34,731 --> 00:19:38,801
HIV, IN A VERY, VERY BROAD WAY, 

484
00:19:38,801 --> 00:19:43,306
WHAT WE CAME TO SORT OF 

485
00:19:43,306 --> 00:19:44,240
APPRECIATE IS THE FACT THAT IN 

486
00:19:44,240 --> 00:19:47,377
THE CASE OF HIV, IT HAS WHAT'S 

487
00:19:47,377 --> 00:19:49,679
CALLED A PRIMARY LATE DOMAIN.

488
00:19:49,679 --> 00:19:51,781
LATE DOMAIN IS A TERM THAT HAD 

489
00:19:51,781 --> 00:19:57,554
BEEN ATTRIBUTED TO THE 

490
00:19:57,554 --> 00:19:58,922
ESSENTIALLY MOTIFS, DOCKING 

491
00:19:58,922 --> 00:20:01,424
SITES, IF YOU WILL, THAT PERMIT 

492
00:20:01,424 --> 00:20:03,793
RETROVIRUSES AND OTHER VIRUSES 

493
00:20:03,793 --> 00:20:06,896
TO RECRUIT MACHINERY THAT WILL 

494
00:20:06,896 --> 00:20:10,533
GET THEM OUT OF THE CELL.

495
00:20:10,533 --> 00:20:13,469
HIV HAD A PRIMARY LATE TO PAIN.

496
00:20:13,469 --> 00:20:16,172
IT WAS TO THIS -- LATE DOMAIN.

497
00:20:16,172 --> 00:20:18,408
IT WAS TO THIS LATE DOMAIN THAT 

498
00:20:18,408 --> 00:20:20,176
WE FOUND TSG101 WAS TARGETED.

499
00:20:20,176 --> 00:20:23,313
THIS LATE DOMAIN IN 

500
00:20:23,313 --> 00:20:26,449
PARTICULAR -- HERE IT IS OVER 

501
00:20:26,449 --> 00:20:33,590
HERE -- IT'S CHARACTERIZED BY 

502
00:20:33,590 --> 00:20:35,959
FOLDING, SOMETIMES PROLINE, WHAT

503
00:20:35,959 --> 00:20:38,695
WE CALL A P/SAP MOTIF.

504
00:20:38,695 --> 00:20:39,796
ANYBODY WHO WORKS IN HIV KNOWS 

505
00:20:39,796 --> 00:20:42,565
IF YOU WERE TO MAKE A MUTATION 

506
00:20:42,565 --> 00:20:45,435
THAT WAS TO DISABLE THE P/SAP 

507
00:20:45,435 --> 00:20:47,170
MOTIF, WELL, DEPENDING ON THE 

508
00:20:47,170 --> 00:20:50,039
CELL TYPE YOU'RE IN, THE VIRUS 

509
00:20:50,039 --> 00:20:51,140
IS DEFINITELY IMPAIRED WITH 

510
00:20:51,140 --> 00:20:52,809
RESPECT TO MAKING VIRAL 

511
00:20:52,809 --> 00:20:54,310
PARTICLES THAT CAN GET OUT OF 

512
00:20:54,310 --> 00:20:56,346
THE CELL, BUT IT CAN STILL MAKE 

513
00:20:56,346 --> 00:20:58,414
VIRAL PARTICLES THAT CAN GET OUT

514
00:20:58,414 --> 00:20:59,549
OF THE CELL.

515
00:20:59,549 --> 00:21:02,151
AND THAT'S BECAUSE IT HAS WHAT 

516
00:21:02,151 --> 00:21:04,554
WE CALL A SECONDARY LATE DOMAIN.

517
00:21:04,554 --> 00:21:08,524
NOW, THIS SECONDARY LATE DOMAIN,

518
00:21:08,524 --> 00:21:11,894
L DOMAIN 2, THIS ONE CAN TAKE 

519
00:21:11,894 --> 00:21:12,528
MANY FLAVORS.

520
00:21:12,528 --> 00:21:15,898
IN THE CASE OF HIV, IT'S 

521
00:21:15,898 --> 00:21:20,136
ESSENTIALLY A LEUCINE YXXL 

522
00:21:20,136 --> 00:21:20,737
AGAIN, RIGHT?

523
00:21:20,737 --> 00:21:22,205
SPECIFICALLY, IN THE CASE OF 

524
00:21:22,205 --> 00:21:24,140
HIV, IT'S AS YOU SEE IT THERE ON

525
00:21:24,140 --> 00:21:29,979
THE BOARD, A LEUCINE TYROSINE 

526
00:21:29,979 --> 00:21:33,616
PROLINE, A LEUCINE SO ON AND SO.

527
00:21:33,616 --> 00:21:36,119
IT CAN ALSO TAKE THE FORM OF 

528
00:21:36,119 --> 00:21:36,319
YXXL.

529
00:21:36,319 --> 00:21:41,324
SO IF YOU KNOCKED OUT THE 

530
00:21:41,324 --> 00:21:44,227
PRIMARY LATE DOMAIN, VIRUS CAN 

531
00:21:44,227 --> 00:21:45,094
STILL GET OUT.

532
00:21:45,094 --> 00:21:47,196
AND THAT'S BECAUSE IT'S 

533
00:21:47,196 --> 00:21:49,332
DEPENDING ON ITS SECONDARY LATE 

534
00:21:49,332 --> 00:21:49,565
DOMAIN.

535
00:21:49,565 --> 00:21:52,335
SO IF YOU NOW KNOCK OUT THE 

536
00:21:52,335 --> 00:21:54,003
SECONDARY LATE DOMAIN AND THE 

537
00:21:54,003 --> 00:21:55,705
PRIMARY LATE DOMAIN, YES, THE 

538
00:21:55,705 --> 00:21:56,839
VIRUS IS IMPAIRED WITH THE 

539
00:21:56,839 --> 00:21:59,142
EFFICIENCY WITH WHICH IT CAN GET

540
00:21:59,142 --> 00:22:02,945
VIRAL PARTICLES OUT, BUT IT ASK 

541
00:22:02,945 --> 00:22:04,981
STILL GET VIRAL PARTICLES OUT 

542
00:22:04,981 --> 00:22:07,316
AND THAT'S BECAUSE OF ITS 

543
00:22:07,316 --> 00:22:08,751
TERTIARY LATE DOMAIN.

544
00:22:08,751 --> 00:22:10,319
IT'S A LATE DOMAIN WHOSE 

545
00:22:10,319 --> 00:22:12,955
STRUCTURE IS NOT SO CLEAR, BUT 

546
00:22:12,955 --> 00:22:15,558
IT IS ONE THAT FUNCTIONS WHEN, 

547
00:22:15,558 --> 00:22:20,096
IN FACT, THE OTHER TWO ARE 

548
00:22:20,096 --> 00:22:20,363
DISABLED.

549
00:22:20,363 --> 00:22:25,001
NOW, IN THIS KIND OF 

550
00:22:25,001 --> 00:22:26,402
RELENTLESSNESS WITH RESPECT TO 

551
00:22:26,402 --> 00:22:27,036
REDUNDANCY OF MECHANISM FOR 

552
00:22:27,036 --> 00:22:28,271
GETTING OUT, ONE OF THE 

553
00:22:28,271 --> 00:22:29,639
QUESTIONS I ASKED MYSELF WAS, 

554
00:22:29,639 --> 00:22:32,942
WELL, YES, IT IS A REDUNDANT 

555
00:22:32,942 --> 00:22:35,044
MECHANISM TO GET VIRUS OUT OF 

556
00:22:35,044 --> 00:22:37,380
CELLS AND, YES, IT MAKES SENSE 

557
00:22:37,380 --> 00:22:39,082
THAT A VIRUS MIGHT WANT TO DO 

558
00:22:39,082 --> 00:22:42,652
THAT, BUT IN POINT OF FACT, 

559
00:22:42,652 --> 00:22:45,388
THERE WAS NOT AT THE TIME AND 

560
00:22:45,388 --> 00:22:48,024
STILL TODAY THERE'S NOT -- I DO 

561
00:22:48,024 --> 00:22:51,360
NOT KNOW OF ANY HIV VARIANT THAT

562
00:22:51,360 --> 00:22:52,395
HAD BEEN ISOLATED GLOBALLY WHERE

563
00:22:52,395 --> 00:22:54,230
THERE'S A MUTATION IN THE 

564
00:22:54,230 --> 00:22:55,598
PRIMARY LATE DOMAIN.

565
00:22:55,598 --> 00:22:56,833
IN OTHER WORDS, THE VIRUS SAYS, 

566
00:22:56,833 --> 00:22:59,168
I'M NOT DOING THAT, I'M NOT 

567
00:22:59,168 --> 00:23:00,536
MESSING WITH THAT, ALL RIGHT?

568
00:23:00,536 --> 00:23:02,004
BUT IN THE LABORATORY, YOU 

569
00:23:02,004 --> 00:23:06,309
CERTAINLY CAN DO IT, AND SO 

570
00:23:06,309 --> 00:23:07,343
OBVIOUSLY IT ALWAYS WAS A 

571
00:23:07,343 --> 00:23:11,581
NAGGING THOUGHT TO US THAT THIS 

572
00:23:11,581 --> 00:23:13,416
CAN'T SIMPLY BE FOR -- HAVING 

573
00:23:13,416 --> 00:23:15,451
THESE LATE DOMAINS CAN'T SIMPLY 

574
00:23:15,451 --> 00:23:17,687
BE FOR THE QUESTION OF 

575
00:23:17,687 --> 00:23:18,955
REDUNDANCY BECAUSE WE HAVEN'T 

576
00:23:18,955 --> 00:23:21,691
GOTTEN A VARIANT IN A NATURAL 

577
00:23:21,691 --> 00:23:22,992
VARIANT YET.

578
00:23:22,992 --> 00:23:27,663
SO IF THERE'S A REASON THAT THE 

579
00:23:27,663 --> 00:23:29,565
VIRUS SHOULD BOTHER TO GO AND 

580
00:23:29,565 --> 00:23:31,434
MAKE THIS SYSTEM TO REDUNDANT.

581
00:23:31,434 --> 00:23:32,702
ONCE AGAIN, ANOTHER THOUGHT 

582
00:23:32,702 --> 00:23:33,736
LURKING IN THE BACK OF THE HEAD 

583
00:23:33,736 --> 00:23:36,906
WAS THAT IN THE CASE OF HIV, IT 

584
00:23:36,906 --> 00:23:37,840
RECRUITS TSG101.

585
00:23:37,840 --> 00:23:41,210
IN THE CASE OF THE AVIAN 

586
00:23:41,210 --> 00:23:44,447
RETROVIRUS, IT RECRUITS THE -- 

587
00:23:44,447 --> 00:23:46,182
THROUGH THE PY MOTIF PROTEINS 

588
00:23:46,182 --> 00:23:51,287
THAT BELONG TO A FAMILY OF 

589
00:23:51,287 --> 00:24:00,129
UBIQUITIN E3 LIGASES, NEDD4 AND 

590
00:24:00,129 --> 00:24:01,497
OTHER MEMBERS OF THE FAMILY AND 

591
00:24:01,497 --> 00:24:04,967
IN THE CASE OF TSG101, IT HAS 

592
00:24:04,967 --> 00:24:09,772
BOTH A P/SAP MOTIF AND WE'LL 

593
00:24:09,772 --> 00:24:13,109
CALL IT A SECONDARY LATE DOMAIN 

594
00:24:13,109 --> 00:24:14,143
MOTIF AND IT ACTUALLY 

595
00:24:14,143 --> 00:24:15,411
UTILIZING -- I'M SORRY, IT MIGHT

596
00:24:15,411 --> 00:24:18,080
HAVE ALL THREE, BUT IT 

597
00:24:18,080 --> 00:24:21,684
DEFINITELY HAS THE P/SAP AND THE

598
00:24:21,684 --> 00:24:25,655
PY AND IN THE FACE OF TSG101, IT

599
00:24:25,655 --> 00:24:26,189
ACTUALLY UTILIZES MOST.

600
00:24:26,189 --> 00:24:30,493
SO THIS STIMULATED THIS THOUGHT 

601
00:24:30,493 --> 00:24:33,095
PROCESS TO ASK ABOUT THE 

602
00:24:33,095 --> 00:24:38,034
POSSIBILITY THAT TSG AND NEDD4 

603
00:24:38,034 --> 00:24:39,836
MIGHT ACTUALLY INTERACT WITH 

604
00:24:39,836 --> 00:24:41,671
EACH OTHER SO THAT NO MATTER 

605
00:24:41,671 --> 00:24:43,005
WHAT DOCKING SITE THE VIRUS 

606
00:24:43,005 --> 00:24:45,508
MIGHT USE, WHETHER IT'S HIV OR 

607
00:24:45,508 --> 00:24:47,043
SOME OTHER VIRUS, THAT EVEN 

608
00:24:47,043 --> 00:24:50,479
THOUGH THEY MAY RECRUIT DIRECTLY

609
00:24:50,479 --> 00:24:51,614
TSG OR THEY MIGHT RECRUIT 

610
00:24:51,614 --> 00:24:57,453
DIRECTLY NEDD4, IF TSG AND NEDD4

611
00:24:57,453 --> 00:24:59,422
INTERACT, THAT COMPLEX WAS GOING

612
00:24:59,422 --> 00:25:00,690
TO BE CALLED.

613
00:25:00,690 --> 00:25:02,592
BACK TO THAT -- MEANWHILE, 

614
00:25:02,592 --> 00:25:04,193
BECAUSE IT TURNS OUT, WHETHER 

615
00:25:04,193 --> 00:25:08,564
HIV OR ANY OTHER VIRUS RECRUITS 

616
00:25:08,564 --> 00:25:10,867
TSG101 DIRECTLY, TSG101 AND 

617
00:25:10,867 --> 00:25:12,401
NEDD4 INTERACT IN SOLUTION AND 

618
00:25:12,401 --> 00:25:15,504
SO THE COMPLEX IS BEING BROUGHT 

619
00:25:15,504 --> 00:25:16,105
TO BEAR.

620
00:25:16,105 --> 00:25:17,473
SO THE KEY PLAYERS THAT WE'RE 

621
00:25:17,473 --> 00:25:20,676
GOING TO TALK ABOUT TODAY ARE 

622
00:25:20,676 --> 00:25:22,178
THE TSG101 PROTEIN.

623
00:25:22,178 --> 00:25:28,584
IT HAS A -- WHAT'S CALLED A 

624
00:25:28,584 --> 00:25:31,821
TERMINAL UBIQUITIN E2 VARIANT OR

625
00:25:31,821 --> 00:25:35,024
UEV DOMAIN.

626
00:25:35,024 --> 00:25:39,795
IT'S CALLED UBIQUITIN E2 VARIANT

627
00:25:39,795 --> 00:25:41,163
BECAUSE IT LOOKS LIKE IT'S A 

628
00:25:41,163 --> 00:25:46,168
STRUCTURAL COMMON WITH THE 

629
00:25:46,168 --> 00:25:46,402
ENZYMES.

630
00:25:46,402 --> 00:25:49,005
THESE ARE UBIQUITIN ENZYMES AND 

631
00:25:49,005 --> 00:25:51,007
THEN IT HAS THE DOWNSTREAM 

632
00:25:51,007 --> 00:25:54,076
DOMAINS THAT ARE VERY IMPORTANT 

633
00:25:54,076 --> 00:25:58,814
FOR IT TO BE ABLE TO READ 

634
00:25:58,814 --> 00:26:04,654
DOWNSTREAM FACTORS THAT ARE IN 

635
00:26:04,654 --> 00:26:08,724
ESCRT-II AND ESCRT-III TO WHERE 

636
00:26:08,724 --> 00:26:09,358
THE TSG101 IS.

637
00:26:09,358 --> 00:26:11,193
AND ONE OF THE THINGS THAT I 

638
00:26:11,193 --> 00:26:14,063
THINK YOU KNOW OR SHOULD 

639
00:26:14,063 --> 00:26:16,832
APPRECIATE IS THAT ESCRT-III 

640
00:26:16,832 --> 00:26:18,034
HOUSES MACHINERY THAT'S CRITICAL

641
00:26:18,034 --> 00:26:25,141
FOR REMODELING MEMBRANE AND 

642
00:26:25,141 --> 00:26:25,875
CYTOSKEL

643
00:26:25,875 --> 00:26:27,243
CYTOSKELETON, AND SO WHEN TSG101

644
00:26:27,243 --> 00:26:30,079
RECRUITS THAT BY VIRTUE OF ITS 

645
00:26:30,079 --> 00:26:33,749
ABILITY TO INTERACT WITH 

646
00:26:33,749 --> 00:26:37,086
ESCRT-II AND IN TURN ESCRT-III, 

647
00:26:37,086 --> 00:26:39,722
IT COMPLETES MACHINERY TO THE 

648
00:26:39,722 --> 00:26:42,892
VIRAL ASSEMBLY SITE AND PROMOTE 

649
00:26:42,892 --> 00:26:43,259
BUDDING.

650
00:26:43,259 --> 00:26:45,394
SO TSG IS A MAJOR PLAYER WITH 

651
00:26:45,394 --> 00:26:48,497
RESPECT TO EXIT.

652
00:26:48,497 --> 00:26:51,667
THEN, WE LOOK AT LATE DOMAIN 

653
00:26:51,667 --> 00:26:52,301
NUMBER 1.

654
00:26:52,301 --> 00:26:57,306
THE LATE DOMAIN NUMBER 1, ITS 

655
00:26:57,306 --> 00:26:58,474
FAVORITE PARTNER IS ACTUALLY A 

656
00:26:58,474 --> 00:27:02,545
MEMBER OF THE NEDD4 FAMILY 

657
00:27:02,545 --> 00:27:06,882
CALLED NEDD4L FOR LONG.

658
00:27:06,882 --> 00:27:10,753
IT CAN SOMETIMES BE CALLED 

659
00:27:10,753 --> 00:27:11,921
NEDD4-2.

660
00:27:11,921 --> 00:27:16,959
THE LATE DOMAIN 2 WILL RECRUIT 

661
00:27:16,959 --> 00:27:24,033
AND ADOPT A PROTEIN, LE 

662
00:27:24,033 --> 00:27:24,600
CHÂTELIER'S.

663
00:27:24,600 --> 00:27:26,068
THIS LE CHÂTELIER'S PROTEIN WILL

664
00:27:26,068 --> 00:27:28,804
RECRUIT ANOTHER MEMBER OF THE 

665
00:27:28,804 --> 00:27:33,843
NEDD4 FAMILY AND THAT IS 

666
00:27:33,843 --> 00:27:37,546
NEDD4-1, AND THEN THE LATE 

667
00:27:37,546 --> 00:27:42,218
DOMAIN 3 WILL RECRUIT ANOTHER 

668
00:27:42,218 --> 00:27:45,521
ADAPTOR PROTEIN AND THAT ADAPTOR

669
00:27:45,521 --> 00:27:49,158
PROTEIN CAN THEN BIND NEDD4-L.

670
00:27:49,158 --> 00:27:54,597
SO THE VIRUS HAS THE ABILITY TO 

671
00:27:54,597 --> 00:27:57,066
BRING TO ITS VICINITY, 

672
00:27:57,066 --> 00:28:01,537
INDIRECTLY THROUGH TSG, NEDD4, 

673
00:28:01,537 --> 00:28:10,546
THIS ISOFORM LONG THROUGH ALI 

674
00:28:10,546 --> 00:28:13,582
LICLIX,

675
00:28:13,582 --> 00:28:16,719
NEDD4-L AND ANOTHER ISOFORM OF 

676
00:28:16,719 --> 00:28:17,953
NEDD4 ABLE TO FIND DIRECTLY.

677
00:28:17,953 --> 00:28:24,960
SO ONCE AGAIN, RELENTLESS AND 

678
00:28:24,960 --> 00:28:25,561
REDUND

679
00:28:25,561 --> 00:28:25,828
REDUNDANT.

680
00:28:25,828 --> 00:28:32,201
NOW -- OOPS, I HAD A FREEZE.

681
00:28:32,201 --> 00:28:33,335
I'M NOT GETTING THIS TO MOVE 

682
00:28:33,335 --> 00:28:37,673
FORWARD.

683
00:28:37,673 --> 00:28:40,843
SO I NEED A LITTLE -- OH, GOOD, 

684
00:28:40,843 --> 00:28:41,143
THANK YOU.

685
00:28:41,143 --> 00:28:43,846
ALL RIGHT.

686
00:28:43,846 --> 00:28:48,017
SO AS I SAID, THE NEDD4 OF 

687
00:28:48,017 --> 00:28:51,854
UBIQUITIN E3 LIGASES IS PRETTY 

688
00:28:51,854 --> 00:28:52,221
LARGE.

689
00:28:52,221 --> 00:28:54,256
THERE'S SEVERAL, SEVERAL 

690
00:28:54,256 --> 00:28:54,490
MEMBERS.

691
00:28:54,490 --> 00:28:57,726
AND TO JUST REVIEW FOR YOU, THIS

692
00:28:57,726 --> 00:28:59,829
FAMILY, WHAT THIS FAMILY DOES IS

693
00:28:59,829 --> 00:29:01,964
ACTUALLY MODIFY PROTEINS BY 

694
00:29:01,964 --> 00:29:04,133
ADDING ON TO THEM UBIQUITIN.

695
00:29:04,133 --> 00:29:05,768
AND THERE ARE MANY DIFFERENT 

696
00:29:05,768 --> 00:29:09,672
WAYS THAT IT CAN ADD UBIQUITIN.

697
00:29:09,672 --> 00:29:12,274
IT CAN ADD JUST ONE UBIQUITIN, 

698
00:29:12,274 --> 00:29:15,211
TYPICALLY, TO A LYCINE.

699
00:29:15,211 --> 00:29:17,913
IT CAN ADD A CHAIN, COULD ADD 

700
00:29:17,913 --> 00:29:18,914
UBIQUITIN AS A CHAIN.

701
00:29:18,914 --> 00:29:21,650
IT COULD ADD UBIQUITIN AS A 

702
00:29:21,650 --> 00:29:21,851
DIMER.

703
00:29:21,851 --> 00:29:25,654
IT COULD ADD UBIQUITIN AS A 

704
00:29:25,654 --> 00:29:27,857
BRANCHED COMPLEX AND COULD ADD 

705
00:29:27,857 --> 00:29:29,058
UBIQUITIN TO MORE THAN ONE SITE 

706
00:29:29,058 --> 00:29:30,826
OF THE PROTEIN.

707
00:29:30,826 --> 00:29:33,496
ALL OF THESE DIFFERENT 

708
00:29:33,496 --> 00:29:35,097
VARIATIONS COMPRISE UBIQUITIN 

709
00:29:35,097 --> 00:29:38,167
SIGNALING, AND THAT SIGNAL IS 

710
00:29:38,167 --> 00:29:40,536
READ BY CELLS TO MANY CERTAIN 

711
00:29:40,536 --> 00:29:41,604
THINGS.

712
00:29:41,604 --> 00:29:44,406
SOME OF THEM MEAN TAKE THIS KARG

713
00:29:44,406 --> 00:29:46,375
FOR TO THE GARBAGE PAIL.

714
00:29:46,375 --> 00:29:48,811
IF THERE'S A UBIQUITIN MACHINE, 

715
00:29:48,811 --> 00:29:52,648
IT SAYS SEND ME TO THE LYSOSOME 

716
00:29:52,648 --> 00:29:57,286
OR THE PROTEO SOME FOR 

717
00:29:57,286 --> 00:29:57,586
DEGRADATION.

718
00:29:57,586 --> 00:29:59,655
IF IT HAS -- IT SAYS TAKE ME 

719
00:29:59,655 --> 00:30:01,023
SOMEWHERE IN THE CELL.

720
00:30:01,023 --> 00:30:04,994
SORT MESO THAT I END UP IN SOME 

721
00:30:04,994 --> 00:30:06,195
LOCATION.

722
00:30:06,195 --> 00:30:09,665
AND THE PROCESS BY WHICH THE 

723
00:30:09,665 --> 00:30:11,500
UBIQUITIN INTERACTS WITH ANOTHER

724
00:30:11,500 --> 00:30:14,637
UBIQUITIN MOLECULE CAN ALSO VARY

725
00:30:14,637 --> 00:30:17,406
AND THAT ALSO IS BEARING A 

726
00:30:17,406 --> 00:30:19,909
SIGNAL TO TREAT THE CARGO IN A 

727
00:30:19,909 --> 00:30:21,677
CERTAIN WAY.

728
00:30:21,677 --> 00:30:23,846
SO WHAT HAPPENS IS, THESE 

729
00:30:23,846 --> 00:30:26,382
ENZYMES SEE UBIQUITIN, THEY WORK

730
00:30:26,382 --> 00:30:29,485
WITH WHAT'S CALLED E1 ENZYMES.

731
00:30:29,485 --> 00:30:30,953
E1 ENZYMES ACTIVATE THE 

732
00:30:30,953 --> 00:30:31,820
UBIQUITIN.

733
00:30:31,820 --> 00:30:34,957
THAT ENABLES THE UBIQUITIN TO BE

734
00:30:34,957 --> 00:30:39,828
TRANSFERRED TO WHAT'S CALLED N 

735
00:30:39,828 --> 00:30:43,199
E2 ENZYME AND THE E2 ENZYME IS A

736
00:30:43,199 --> 00:30:44,967
UBIQUITIN CONJUGATING ENZYME.

737
00:30:44,967 --> 00:30:47,937
THE UBIQUITIN CONJUGATING ENZYME

738
00:30:47,937 --> 00:30:50,372
IS WHAT I REFERRED TO BEFORE AS 

739
00:30:50,372 --> 00:31:00,783
BEING STRUCTURAL HOMOLOGOUS TO 

740
00:31:00,783 --> 00:31:03,319
THE TSG101 TO MAIM.

741
00:31:03,319 --> 00:31:05,354
THESE WERE PASS THE WAITING 

742
00:31:05,354 --> 00:31:07,189
UBIQUITIN TO AN E3 ENZYME AND 

743
00:31:07,189 --> 00:31:10,226
WHEN IT DOES THAT, IT TRANSFERS 

744
00:31:10,226 --> 00:31:13,662
THE UBIQUITIN WAITING TO ASSIST 

745
00:31:13,662 --> 00:31:16,332
IN THE ACTIVE SITE OF THE ENZYME

746
00:31:16,332 --> 00:31:20,169
AND THEN THAT IS ABLE TO PICK A 

747
00:31:20,169 --> 00:31:23,072
SUBSTRATE AND IT PICKS A 

748
00:31:23,072 --> 00:31:23,906
SUBSTRATE AND IT THEN WILL 

749
00:31:23,906 --> 00:31:25,608
MODIFY THAT SUBSTRATE WITH 

750
00:31:25,608 --> 00:31:25,874
UBIQUITIN.

751
00:31:25,874 --> 00:31:29,445
THE WAY IT PICKS THE SUBSTRATE 

752
00:31:29,445 --> 00:31:31,947
IS BY VIRTUE OF A DOMAIN WITHIN 

753
00:31:31,947 --> 00:31:36,385
THE E3 LIGASE THAT IS SPECIALLY 

754
00:31:36,385 --> 00:31:38,487
DESIGNED FOR SUBSTRATE 

755
00:31:38,487 --> 00:31:39,855
RECOGNITION.

756
00:31:39,855 --> 00:31:42,358
TYPICALLY SUBSTRATES WILL HAVE A

757
00:31:42,358 --> 00:31:43,826
PY MOTIF.

758
00:31:43,826 --> 00:31:45,060
NOW, EACH VIRUS HAS ITS 

759
00:31:45,060 --> 00:31:45,394
FAVORITE.

760
00:31:45,394 --> 00:31:47,529
IN THE CASE OF HIV, I'VE 

761
00:31:47,529 --> 00:31:49,565
MENTIONED THREE THAT ARE ITS 

762
00:31:49,565 --> 00:31:50,966
FAVORITE, BUT IF YOU WERE 

763
00:31:50,966 --> 00:32:00,209
DEALING WITH, SAY FOR EXAMPLE, 

764
00:32:00,209 --> 00:32:06,482
HCLV-1, IT WOULDN'T BE NEDD4-1 

765
00:32:06,482 --> 00:32:08,517
OR 2 OR LONG.

766
00:32:08,517 --> 00:32:11,887
IT WOULD BE WWP1.

767
00:32:11,887 --> 00:32:14,323
THAT'S A FAVORITE.

768
00:32:14,323 --> 00:32:15,824
OR IT WOULD BE SMURF, ANOTHER 

769
00:32:15,824 --> 00:32:16,392
FAVORITE.

770
00:32:16,392 --> 00:32:17,893
THE REASON FOR THAT IS REALLY 

771
00:32:17,893 --> 00:32:20,029
INTERESTING TO ME BECAUSE HIV 

772
00:32:20,029 --> 00:32:21,163
DOESN'T LIKE THOSE.

773
00:32:21,163 --> 00:32:23,132
SO EACH VIRUS HAS ITS FAVORITE 

774
00:32:23,132 --> 00:32:26,635
AND EACH CASE, YOU'RE CALLING 

775
00:32:26,635 --> 00:32:31,707
DIFFERENT CONNECTORS TO THE 

776
00:32:31,707 --> 00:32:33,208
ASSEMBLIAGE THROUGH THIS 

777
00:32:33,208 --> 00:32:35,744
PARTICULAR PARTNER.

778
00:32:35,744 --> 00:32:37,680
NOW, SO I SAID IT BEFORE.

779
00:32:37,680 --> 00:32:41,350
LATE DOMAINS, THEY FACILITY 

780
00:32:41,350 --> 00:32:42,251
CELL-FREE VIRUS EXIT.

781
00:32:42,251 --> 00:32:46,255
AND SO IF YOU'RE DEALING WITH, 

782
00:32:46,255 --> 00:32:50,893
SAY, NEDD4L, THEN THAT ENZYME BY

783
00:32:50,893 --> 00:32:55,397
VIRTUE OF ITS WW DOMAIN CAN BIND

784
00:32:55,397 --> 00:32:57,966
THE OTHER PARTNERS IN THE CELL.

785
00:32:57,966 --> 00:33:00,469
AND THIS IS JUST A COLLECTION OF

786
00:33:00,469 --> 00:33:03,405
SOME OF THE PROTEINS THAT ARE 

787
00:33:03,405 --> 00:33:06,742
ABLE TO INTERACT WITH NEDD4L, 

788
00:33:06,742 --> 00:33:09,078
AND YOU CAN SEE THAT AMOT IS ONE

789
00:33:09,078 --> 00:33:10,446
OF THEM.

790
00:33:10,446 --> 00:33:14,516
AMOT WAS THE ADAPTOR PROTEIN 

791
00:33:14,516 --> 00:33:21,757
THAT LEADS NEDD4 TO THE LATE 

792
00:33:21,757 --> 00:33:22,224
DOMAIN 3.

793
00:33:22,224 --> 00:33:26,428
IF YOU WERE TALKING ABOUT, SAY, 

794
00:33:26,428 --> 00:33:28,764
NEDD4-1, ITS WW DOMAINS ARE 

795
00:33:28,764 --> 00:33:30,632
DIFFERENT AND THE COLLECTION OF 

796
00:33:30,632 --> 00:33:33,736
PROTEINS THAT IT CALLS ARE 

797
00:33:33,736 --> 00:33:35,003
DIFFERENT, AND HERE ACTUALLY, 

798
00:33:35,003 --> 00:33:36,839
ONE OF THESE PROTEINS WOULD THEN

799
00:33:36,839 --> 00:33:41,744
BE AN ADAPTOR THAT WOULD PERMIT 

800
00:33:41,744 --> 00:33:46,382
ALIX TO CONNECT TO NEDD4-1 AND 

801
00:33:46,382 --> 00:33:46,849
SO ON.

802
00:33:46,849 --> 00:33:49,385
SO YOU CAN SEE BY HAVING THIS 

803
00:33:49,385 --> 00:33:51,954
KIND OF RELATIONSHIP, THE VIRUS 

804
00:33:51,954 --> 00:33:55,657
IS ABLE TO CALL PROTEINS THAT 

805
00:33:55,657 --> 00:33:59,661
ARE NOT DIRECTLY ASSOCIATED WITH

806
00:33:59,661 --> 00:34:03,632
THE GAG PER SE TO THE ASSEMBLAGE

807
00:34:03,632 --> 00:34:05,734
FOR VARIOUS PURPOSES, WHICH 

808
00:34:05,734 --> 00:34:09,271
WE'LL ALLUDE IN A MINUTE.

809
00:34:09,271 --> 00:34:09,671
OKAY.

810
00:34:09,671 --> 00:34:14,276
SO IRRESPECTIVE OF GAG, I SAID 

811
00:34:14,276 --> 00:34:16,678
BEFORE THAT THE TSG101 UEV 

812
00:34:16,678 --> 00:34:22,551
DOMAIN AND THE NEDD4 HECT DOMAIN

813
00:34:22,551 --> 00:34:26,655
ARE ABLE TO INTERACT AND FOR 

814
00:34:26,655 --> 00:34:27,890
NMR, WE CAN IDENTIFY THOSE 

815
00:34:27,890 --> 00:34:30,459
REGIONS IN BOTH THE UEV DOMAIN 

816
00:34:30,459 --> 00:34:32,194
AND THE HECT DOMAIN WHERE THIS 

817
00:34:32,194 --> 00:34:33,495
INTERACTION OCCURS.

818
00:34:33,495 --> 00:34:34,763
I CAN TELL YOU THAT THIS 

819
00:34:34,763 --> 00:34:35,998
INTERACTION IS WEAK.

820
00:34:35,998 --> 00:34:38,467
IT'S LIKE THE INTERACTION OF 

821
00:34:38,467 --> 00:34:41,170
UBIQUITIN.

822
00:34:41,170 --> 00:34:42,371
UBIQUITIN'S INTERACTIONS TEND TO

823
00:34:42,371 --> 00:34:44,706
NOT BE HIGH AFFINITY BECAUSE 

824
00:34:44,706 --> 00:34:45,641
THEY WANT TO BE MOBILE.

825
00:34:45,641 --> 00:34:48,010
THEY WANT TO BE ABLE TO BE ON 

826
00:34:48,010 --> 00:34:51,847
AND OFF IN TERMS OF CONTROLLING 

827
00:34:51,847 --> 00:34:52,514
SIGNALING.

828
00:34:52,514 --> 00:34:55,884
IN THE CASE OF THE TSG UEV, WE 

829
00:34:55,884 --> 00:35:00,122
KNOW THAT ONE REGION IN NEDD4 

830
00:35:00,122 --> 00:35:02,691
THAT INTERACTS WITH IS THIS 

831
00:35:02,691 --> 00:35:05,194
REGION HERE CALLED ALPHA 

832
00:35:05,194 --> 00:35:05,661
HELIX-1.

833
00:35:05,661 --> 00:35:07,563
WE KNOW THAT THERE'S ANOTHER 

834
00:35:07,563 --> 00:35:15,137
REGION OF NEDD4, SO NEDD4 HAS --

835
00:35:15,137 --> 00:35:18,574
AS PART OF ITS ENZYMATIC DOMAIN,

836
00:35:18,574 --> 00:35:21,343
IT'S HECT DOMAIN, IT HAS AN 

837
00:35:21,343 --> 00:35:23,946
N-LOBBY AND A C-LOBE.

838
00:35:23,946 --> 00:35:28,083
THE C-LOBE IS WHERE THE PROTEIN 

839
00:35:28,083 --> 00:35:29,685
THAT WILL RECEIVE THE UBIQUITIN 

840
00:35:29,685 --> 00:35:32,921
FROM THE E2 ENZYME WOULD SIT, 

841
00:35:32,921 --> 00:35:35,257
AND THIS N-LOBE IS ACTUALLY 

842
00:35:35,257 --> 00:35:38,093
CONSIDERED LIKE A WAITING 

843
00:35:38,093 --> 00:35:41,063
STATION, MEANING THAT UBIQUITIN 

844
00:35:41,063 --> 00:35:42,297
MOLECULES THAT ARE DESTINED TO 

845
00:35:42,297 --> 00:35:45,634
BE, SAY, ADDED TO A UBIQUITIN 

846
00:35:45,634 --> 00:35:47,603
CHAIN, THEY WILL SIT IN THIS 

847
00:35:47,603 --> 00:35:50,706
WAITING ROOM OVER IN THE N-LOBE.

848
00:35:50,706 --> 00:35:54,543
SO THAT AS YOU HAVE AN E2 

849
00:35:54,543 --> 00:35:56,778
TRANSFERRING A UBIQUITIN TO AN 

850
00:35:56,778 --> 00:35:59,014
E3 AND THEN E3 TRANSFERRING IT 

851
00:35:59,014 --> 00:36:03,585
TO A SUBSTRATE, UBIQUITIN IN THE

852
00:36:03,585 --> 00:36:08,524
WAITING ROOM WILL THEN JOIN TO 

853
00:36:08,524 --> 00:36:11,393
THE UBIQUITIN THAT'S ON THE 

854
00:36:11,393 --> 00:36:12,661
SUBSTRATE AND MAKE SOME KIND OF 

855
00:36:12,661 --> 00:36:13,695
UBIQUITIN CASCADE THAT IS THE 

856
00:36:13,695 --> 00:36:16,064
SIGNAL.

857
00:36:16,064 --> 00:36:16,999
ALL RIGHT.

858
00:36:16,999 --> 00:36:20,936
SO ONE OF THE THINGS THAT WE DID

859
00:36:20,936 --> 00:36:26,408
WAS, WE LOOKED AT THE RESIDUES 

860
00:36:26,408 --> 00:36:30,712
THAT WERE IN UEV PROXIMITY, AND 

861
00:36:30,712 --> 00:36:34,449
WE MUTATED THOSE RESIDUES, AND 

862
00:36:34,449 --> 00:36:38,020
WHAT WE FOUND IS THAT IN A 

863
00:36:38,020 --> 00:36:39,488
CERTAIN REGION OF THE HECT 

864
00:36:39,488 --> 00:36:43,992
ENZYMATIC DOMAIN IN NEDD4, 

865
00:36:43,992 --> 00:36:47,029
MUTATIONS IN THIS ALPHA HELIX-1 

866
00:36:47,029 --> 00:36:53,669
REGION, WHICH IS IN UEV 

867
00:36:53,669 --> 00:36:56,405
PROXIMITY, THAT INTERFERED, 

868
00:36:56,405 --> 00:36:56,972
MINIMIZED, DIMINISHED THE 

869
00:36:56,972 --> 00:37:01,343
ABILITY OF NEDD4, THE WILD-TYPE 

870
00:37:01,343 --> 00:37:02,344
NEDD4 TO RESCUE.

871
00:37:02,344 --> 00:37:04,913
SO YOU'RE DEALING WITH A VIRUS 

872
00:37:04,913 --> 00:37:05,781
WHERE YOU'VE ACTUALLY KNOCKED 

873
00:37:05,781 --> 00:37:08,016
OUT LATE DOMAIN 1 AND LATE 

874
00:37:08,016 --> 00:37:10,152
DOMAIN 2 SO THAT NOW IT IS 

875
00:37:10,152 --> 00:37:13,922
TOTALLY DEPENDENT ON LATE DOMAIN

876
00:37:13,922 --> 00:37:17,392
3 TO BUD OUT OF THE CELL.

877
00:37:17,392 --> 00:37:18,427
AND YOU CAN DEMONSTRATE THAT, 

878
00:37:18,427 --> 00:37:21,730
BECAUSE IF YOU TAKE THAT VIRUS 

879
00:37:21,730 --> 00:37:24,333
WHICH IS IN ONE SELECTIVE 

880
00:37:24,333 --> 00:37:27,336
ANALYSIS AND THE MATERIAL IN THE

881
00:37:27,336 --> 00:37:28,670
SUPERNATANT HAS BEEN SEMI 

882
00:37:28,670 --> 00:37:30,072
PURIFIED, SO WE'RE LOOKING AT 

883
00:37:30,072 --> 00:37:31,840
VIRAL-LIKE PROTEINS, AND THIS IS

884
00:37:31,840 --> 00:37:33,742
HOW MUCH YOU ACTUALLY GET OUT IF

885
00:37:33,742 --> 00:37:37,112
YOU DIDN'T ADD ANYTHING, BUT NOW

886
00:37:37,112 --> 00:37:43,785
IF YOU ADD WILD-TYPE NEDD4L, YOU

887
00:37:43,785 --> 00:37:45,654
CAN RESCUE THE VIRUS.

888
00:37:45,654 --> 00:37:47,489
IF YOU MAKE MUTATION NONTHE 

889
00:37:47,489 --> 00:37:49,858
REGION OF ALPHA HELIX-1 THAT ARE

890
00:37:49,858 --> 00:37:53,261
IN CLOSE PROXIMITY TO THE UEV 

891
00:37:53,261 --> 00:37:54,630
DOMAIN IN TSG, NOW YOU REPAIR 

892
00:37:54,630 --> 00:37:58,133
THE ABILITY OF THE NEDD4 TO 

893
00:37:58,133 --> 00:37:58,600
RESCUE.

894
00:37:58,600 --> 00:38:00,902
NOW, YOU COULD DO THE SAME THING

895
00:38:00,902 --> 00:38:05,140
IN OTHER REGIONS OF NEDD4.

896
00:38:05,140 --> 00:38:07,843
THIS IS ANOTHER PHASE OF THE 

897
00:38:07,843 --> 00:38:10,579
HECT DOMAIN IN NEDD4 WHERE WE 

898
00:38:10,579 --> 00:38:12,447
ENDED UP MAKING MUTATIONS ALL 

899
00:38:12,447 --> 00:38:15,283
OVER THIS DOMAIN, ALL RIGHT.

900
00:38:15,283 --> 00:38:18,020
IT'S GOT THE REGION WHICH IS 

901
00:38:18,020 --> 00:38:23,458
WHERE THE ACTIVE SITE C SISTINE 

902
00:38:23,458 --> 00:38:24,159
WOULD ACTUALLY SIT.

903
00:38:24,159 --> 00:38:27,696
IT'S GOT A REGION WHERE AN E2 

904
00:38:27,696 --> 00:38:28,997
WOULD ACTUALLY SIT WHEN IT'S 

905
00:38:28,997 --> 00:38:30,699
TRANSFERRING THAT UBIQUITIN TO 

906
00:38:30,699 --> 00:38:32,501
THAT CYSTINE.

907
00:38:32,501 --> 00:38:34,202
IT HAS THAT WAITING STATION THAT

908
00:38:34,202 --> 00:38:37,272
I MENTIONED TO YOU BEFORE WHERE 

909
00:38:37,272 --> 00:38:38,940
UBIQUITINS JUST SIT LYING IN 

910
00:38:38,940 --> 00:38:39,408
WAIT.

911
00:38:39,408 --> 00:38:41,710
BUT WHEN YOU MAKE MUTATIONS, 

912
00:38:41,710 --> 00:38:45,414
THESE SITES, THE ONLY SITES THAT

913
00:38:45,414 --> 00:38:48,450
ACTUALLY ENDED UP IMPAIRING THE 

914
00:38:48,450 --> 00:38:51,453
ABILITY TO GET INFECTIOUS VIRUS 

915
00:38:51,453 --> 00:38:53,655
MADE WERE THE SITES I'VE 

916
00:38:53,655 --> 00:38:55,424
HIGHLIGHTED HERE IN RED.

917
00:38:55,424 --> 00:38:58,393
HERE, HERE, AND HERE.

918
00:38:58,393 --> 00:39:02,297
AND WHAT THIS IS, ACTUALLY, IT'S

919
00:39:02,297 --> 00:39:05,033
A REGION THAT'S CALLED THE HINGE

920
00:39:05,033 --> 00:39:05,267
REGION.

921
00:39:05,267 --> 00:39:08,503
IT'S A HINGE REGION BETWEEN THAT

922
00:39:08,503 --> 00:39:12,074
C-LOBE AND N-LOBE AND THE WHOLE 

923
00:39:12,074 --> 00:39:15,043
ISSUE OF HOW E3 LIGASES 

924
00:39:15,043 --> 00:39:16,912
CATALYZE, DO WHAT THEY DO, 

925
00:39:16,912 --> 00:39:18,180
UBIQUITIN TRANSFER, IS NOT 

926
00:39:18,180 --> 00:39:22,484
KNOWN, BUT THE CONSENSUS IS THAT

927
00:39:22,484 --> 00:39:24,486
THE HINGE REGION ENABLES 

928
00:39:24,486 --> 00:39:26,555
COMMUNICATION BETWEEN THE N-LOBE

929
00:39:26,555 --> 00:39:32,127
AND THE C-LOBE, AND BY OUR 

930
00:39:32,127 --> 00:39:34,162
FINDING MUTATIONS IN THAT 

931
00:39:34,162 --> 00:39:35,063
REGION, BEING THE ONES THAT 

932
00:39:35,063 --> 00:39:37,265
IMPAIR THE ABILITY TO GET 

933
00:39:37,265 --> 00:39:39,835
INFECTIOUS VIRUS OUT, IT SAYS --

934
00:39:39,835 --> 00:39:42,070
IT SUGGESTS THAT THIS WHOLE 

935
00:39:42,070 --> 00:39:45,674
ACTIVITY OF NEDD4 IS PLAYING AN 

936
00:39:45,674 --> 00:39:47,375
IMPORTANT ROLE IN BEING ABLE TO 

937
00:39:47,375 --> 00:39:49,211
GET INFECTIOUS VIRUS OUT.

938
00:39:49,211 --> 00:39:51,780
NOW, A KEY THING, INFECTIOUS 

939
00:39:51,780 --> 00:39:52,047
VIRUS.

940
00:39:52,047 --> 00:39:55,784
THE REASON I KEEP SAYING 

941
00:39:55,784 --> 00:39:57,152
INFECTIOUS VIRUS IS NONE OF 

942
00:39:57,152 --> 00:39:58,920
THESE MUTATIONS HAD ANY EFFECT 

943
00:39:58,920 --> 00:39:59,755
ON BUDDING.

944
00:39:59,755 --> 00:40:01,223
IN OTHER WORDS, YOU COULD MAKE 

945
00:40:01,223 --> 00:40:04,159
MUTATIONS IN THIS REGION, VIRUS 

946
00:40:04,159 --> 00:40:06,928
COULD GET OUT JUST FINE AS AN 

947
00:40:06,928 --> 00:40:08,263
IMMATURE PARTICLE, BUT IT COULD 

948
00:40:08,263 --> 00:40:09,598
NOT MATURE TO BE INFECTIOUS.

949
00:40:09,598 --> 00:40:11,399
WHAT THAT ESSENTIALLY SAYS IS 

950
00:40:11,399 --> 00:40:15,003
THAT THE VIRUS USES SEVERAL 

951
00:40:15,003 --> 00:40:17,272
PARTS IN RELATIONSHIP WITH NEDD4

952
00:40:17,272 --> 00:40:20,408
TO CONTROL BOTH THE BUDDING AND 

953
00:40:20,408 --> 00:40:25,680
THE MATURATION OF THE VIRUS.

954
00:40:25,680 --> 00:40:27,082
NOW, ONCE AGAIN, THE WHOLE THING

955
00:40:27,082 --> 00:40:29,017
HAS TO DO WITH THE FACT THAT WE 

956
00:40:29,017 --> 00:40:32,521
MADE MUTATIONS IN THE REGIONS OF

957
00:40:32,521 --> 00:40:35,090
THE HECT DOMAIN THAT WERE IN 

958
00:40:35,090 --> 00:40:38,593
CLOSE PROXIMITY BY NMR OF THE 

959
00:40:38,593 --> 00:40:39,761
UEV DOMAIN IN TSG.

960
00:40:39,761 --> 00:40:42,497
IF YOU MAKE MUTATIONS OUTSIDE, 

961
00:40:42,497 --> 00:40:47,903
SOME REGION THAT IS NOT IN UEV 

962
00:40:47,903 --> 00:40:49,671
HECT PROXIMITY, VIRUS COULDN'T 

963
00:40:49,671 --> 00:40:50,205
CARE LESS.

964
00:40:50,205 --> 00:40:51,173
ALL RIGHT.

965
00:40:51,173 --> 00:40:54,309
SO WE SAID, ALL RIGHT, WE KNOW 

966
00:40:54,309 --> 00:40:57,445
THAT ENZYMES LIKE NEDD4, THE 

967
00:40:57,445 --> 00:40:59,281
CELL DOESN'T WANT IT TO BE 

968
00:40:59,281 --> 00:41:01,283
ACTIVE, SO THEY ARE TYPICALLY IN

969
00:41:01,283 --> 00:41:04,085
CELLS IN AUTO INHIBITED STATE, 

970
00:41:04,085 --> 00:41:07,923
AND IN THAT AUTO-INHIBITED 

971
00:41:07,923 --> 00:41:10,392
STATE, TYPICALLY YOU HAVE THE C2

972
00:41:10,392 --> 00:41:13,328
DOMAIN AND THIS IS A DOMAIN THAT

973
00:41:13,328 --> 00:41:17,532
TYPICALLY HELPS TO DECIDE WHERE 

974
00:41:17,532 --> 00:41:19,167
NEDD4 CAN DOCK IN THE CELL.

975
00:41:19,167 --> 00:41:21,336
TYPICALLY IN THE AUTO-INHIBITED 

976
00:41:21,336 --> 00:41:24,306
STATE, THE C2 DOMAIN IS ACTUALLY

977
00:41:24,306 --> 00:41:26,842
IN CONTACT WITH THE HECT DOMAIN.

978
00:41:26,842 --> 00:41:28,176
AND BECAUSE IT'S IN CONTACT WITH

979
00:41:28,176 --> 00:41:30,946
THE HECT DOMAIN, THE HECT DOMAIN

980
00:41:30,946 --> 00:41:32,781
IS NOT IN A POSITION NOW TO BE 

981
00:41:32,781 --> 00:41:33,014
ACTIVE.

982
00:41:33,014 --> 00:41:36,051
ITS ACTIVE SITE IS HIDDEN.

983
00:41:36,051 --> 00:41:38,720
SO WE REASONED THAT MAYBE TSG'S 

984
00:41:38,720 --> 00:41:42,624
ROLE WAS TO ACTUALLY INTERFERE 

985
00:41:42,624 --> 00:41:44,559
WITH THAT C2 HECT INTERACTION 

986
00:41:44,559 --> 00:41:46,328
AND TO TEST THIS, WE TOOK 

987
00:41:46,328 --> 00:41:48,163
ADVANTAGE OF THE FACT THAT 

988
00:41:48,163 --> 00:41:53,735
THERE'S A NEDD4 ISOMER THAT CAN 

989
00:41:53,735 --> 00:41:55,704
BIND DIRECTLY THAT HAD BEEN 

990
00:41:55,704 --> 00:41:59,174
IDENTIFIED EARLIER BY THE SUN 

991
00:41:59,174 --> 00:42:03,712
QUIST AND DOPLINGER LABS.

992
00:42:03,712 --> 00:42:06,214
THIS IS AN ISOMER THAT HAS A BIT

993
00:42:06,214 --> 00:42:09,718
OF THE C2 DOMAIN, NOT ENOUGH TO 

994
00:42:09,718 --> 00:42:12,387
MAKE THE CONTACT WITH THE HECT.

995
00:42:12,387 --> 00:42:14,923
WE ASKED WHETHER IF IN FACT YOU 

996
00:42:14,923 --> 00:42:17,192
NEEDED THOSE RESIDUES IN THE 

997
00:42:17,192 --> 00:42:18,560
HECT DOMAIN FOR STABILIZING THE 

998
00:42:18,560 --> 00:42:23,565
INTERACTION WITH THE UEV DOMAIN,

999
00:42:23,565 --> 00:42:26,167
AND IF IN FACT YOU ACTUALLY USE 

1000
00:42:26,167 --> 00:42:30,772
AN ENZYME, A NEDD4 THAT DOESN'T 

1001
00:42:30,772 --> 00:42:35,644
HAVE THAT C2 DOMAIN, THAT SHOULD

1002
00:42:35,644 --> 00:42:37,379
ALWAYS BE ACTIVE AND SO MAYBE 

1003
00:42:37,379 --> 00:42:38,446
THEN YOU DON'T NEED THE TSG AND 

1004
00:42:38,446 --> 00:42:39,881
WE TESTED THAT IDEA AND THAT'S 

1005
00:42:39,881 --> 00:42:41,483
WHAT'S HERE ON THE SLIDE.

1006
00:42:41,483 --> 00:42:42,951
IN OTHER WORDS, EVEN THOUGH 

1007
00:42:42,951 --> 00:42:48,056
TYPICALLY WITH A REGULAR NEDD4 

1008
00:42:48,056 --> 00:42:51,860
MUTANT, IT WOULD NOT BE ABLE TO 

1009
00:42:51,860 --> 00:42:54,429
RESCUE BUDDING IF IN FACT YOU 

1010
00:42:54,429 --> 00:42:56,731
RESCUE WITH THE ISOFORM THAT'S 

1011
00:42:56,731 --> 00:42:58,900
LACKING THE C2 DOMAIN, THEY ARE 

1012
00:42:58,900 --> 00:43:00,802
ABLE TO RESCUE.

1013
00:43:00,802 --> 00:43:04,105
AND SO THAT THEN GAVE US THE 

1014
00:43:04,105 --> 00:43:06,041
NOTION THAT THE TSG UEV IS 

1015
00:43:06,041 --> 00:43:09,945
ACTUALLY HOLDING THE NEDD4 IN AN

1016
00:43:09,945 --> 00:43:11,880
ACTIVATED STATE.

1017
00:43:11,880 --> 00:43:12,147
ALL RIGHT.

1018
00:43:12,147 --> 00:43:16,351
NOW, ONE OF THE MOST, I THINK, 

1019
00:43:16,351 --> 00:43:18,954
INTERESTING STUDIES THAT I HAD 

1020
00:43:18,954 --> 00:43:20,588
AN OPPORTUNITY TO DO TOOK PLACE 

1021
00:43:20,588 --> 00:43:23,358
WHEN I WAS COLLABORATING WITH 

1022
00:43:23,358 --> 00:43:26,428
THE WISE, THE WOMEN'S 

1023
00:43:26,428 --> 00:43:28,063
INTERAGENCY HIV STUDY GROUP.

1024
00:43:28,063 --> 00:43:29,297
AND THE REASON THAT THAT WAS 

1025
00:43:29,297 --> 00:43:32,467
PARTICULARLY FASCINATING TO ME 

1026
00:43:32,467 --> 00:43:35,770
IS BECAUSE AS BASIC RESEARCHERS,

1027
00:43:35,770 --> 00:43:37,339
WE FORMED SOME HYPOTHESES AND WE

1028
00:43:37,339 --> 00:43:40,842
THINK OF EXPERIMENTS TO TEST THE

1029
00:43:40,842 --> 00:43:41,309
HYPOTHESIS.

1030
00:43:41,309 --> 00:43:44,512
BUT IN THIS PARTICULAR CASE, THE

1031
00:43:44,512 --> 00:43:47,515
WIHS WHEN I JOINED THIS 

1032
00:43:47,515 --> 00:43:49,184
PARTICULAR GROUP WAS 20 

1033
00:43:49,184 --> 00:43:50,418
PHYSICIANS, CLINICIANS, AND WHAT

1034
00:43:50,418 --> 00:43:53,922
THEY DO IS THEY INTERACT WITH 

1035
00:43:53,922 --> 00:43:55,757
PEOPLE LIVING WITH HIV AND THEIR

1036
00:43:55,757 --> 00:43:57,659
PROBLEMS AND THEY TRY TO 

1037
00:43:57,659 --> 00:43:58,760
UNDERSTAND WHAT'S THE NATURE OF 

1038
00:43:58,760 --> 00:44:00,528
THE PROBLEM SO THEY CAN, YOU 

1039
00:44:00,528 --> 00:44:01,663
KNOW, WORK TO SOLVE IT.

1040
00:44:01,663 --> 00:44:05,300
SO AT THE POINT THAT I JOINED 

1041
00:44:05,300 --> 00:44:09,704
IT, THERE WAS THIS SUBSET OF THE

1042
00:44:09,704 --> 00:44:12,774
PARTICIPANTS FOR WHOM, AT THE 

1043
00:44:12,774 --> 00:44:15,710
TIME, VERY FABULOUS ARCH WAS NOT

1044
00:44:15,710 --> 00:44:17,512
WORKING.

1045
00:44:17,512 --> 00:44:19,447
THESE PEOPLE WERE ON THE 

1046
00:44:19,447 --> 00:44:21,950
PROTEASE INHIBITORS WHICH AT 

1047
00:44:21,950 --> 00:44:25,653
THAT TIME WERE THE SAVIOR OF THE

1048
00:44:25,653 --> 00:44:28,089
ART WHEN -- IN THOSE DAYS, AND 

1049
00:44:28,089 --> 00:44:30,558
IT HAD BEEN REALLY WORKING 

1050
00:44:30,558 --> 00:44:31,593
BEAUTIFULLY, BUT FOR THEM, THEY 

1051
00:44:31,593 --> 00:44:33,661
WERE NONRESPONSIVE.

1052
00:44:33,661 --> 00:44:35,597
AND, OF COURSE, THEY KNEW THAT 

1053
00:44:35,597 --> 00:44:37,632
THERE WERE NO RESISTANT 

1054
00:44:37,632 --> 00:44:41,069
MUTATIONS THAT WERE IN THE 

1055
00:44:41,069 --> 00:44:43,838
TARGET OF THE ART.

1056
00:44:43,838 --> 00:44:44,873
THAT'S THE FIRST THING THEY 

1057
00:44:44,873 --> 00:44:45,540
CHECKED FOR.

1058
00:44:45,540 --> 00:44:47,208
SO THEY WERE AT A LOSS OF WHAT 

1059
00:44:47,208 --> 00:44:48,410
COULD BE GOING ON.

1060
00:44:48,410 --> 00:44:51,813
SO THEY PERMITTED US TO ACTUALLY

1061
00:44:51,813 --> 00:44:55,784
ISOLATE VARIANTS FROM THESE 

1062
00:44:55,784 --> 00:44:56,918
PARTICIPANTS, LIST OUT THE 

1063
00:44:56,918 --> 00:44:59,187
REGION THAT WE WERE INTERESTED 

1064
00:44:59,187 --> 00:45:01,423
IN, WHICH WAS GAG THROUGH THE 

1065
00:45:01,423 --> 00:45:03,224
END OF THE PROTEASE CHAIN, AND 

1066
00:45:03,224 --> 00:45:05,393
TO SEQUENCE IT.

1067
00:45:05,393 --> 00:45:09,097
AND WHEN WE DID, WE FOUND THAT 

1068
00:45:09,097 --> 00:45:12,500
THERE WERE MUTATIONS IN WHAT WAS

1069
00:45:12,500 --> 00:45:17,138
IN FACT THE BINDING SITE OF 

1070
00:45:17,138 --> 00:45:17,405
ALIX.

1071
00:45:17,405 --> 00:45:19,140
THAT IS TO SAY, LATE DOMAIN 2, 

1072
00:45:19,140 --> 00:45:21,776
AND THESE MUTATIONS IN AND OF 

1073
00:45:21,776 --> 00:45:22,977
THEMSELVES WERE VERY 

1074
00:45:22,977 --> 00:45:23,445
INTERESTING.

1075
00:45:23,445 --> 00:45:25,947
FIRST OF ALL, WITHIN THE GAG 

1076
00:45:25,947 --> 00:45:29,050
FRAME, THEY WERE NOT IN A VERY 

1077
00:45:29,050 --> 00:45:30,985
CRITICAL TYROSINE RESIDUE, WHICH

1078
00:45:30,985 --> 00:45:35,023
IS THE MAKER OR BREAKER OF ALIX 

1079
00:45:35,023 --> 00:45:35,256
BINDING.

1080
00:45:35,256 --> 00:45:39,461
THEY WERE ACTUALLY IN A HIGHLY 

1081
00:45:39,461 --> 00:45:44,132
CONSERVED SPARING RESIDUE.

1082
00:45:44,132 --> 00:45:45,667
HERE.

1083
00:45:45,667 --> 00:45:47,669
AND THE MUTATIONS CHANGED THIS 

1084
00:45:47,669 --> 00:45:50,805
TO NOT JUST ANYTHING, BUT TO 

1085
00:45:50,805 --> 00:45:57,879
EITHER ALAMINE OR PHETYLALMA 

1086
00:45:57,879 --> 00:45:58,213
MEAN.

1087
00:45:58,213 --> 00:46:01,549
THIS IS A REGION IN THE GAG 

1088
00:46:01,549 --> 00:46:02,550
PRECURSOR, WHICH IS WHAT'S 

1089
00:46:02,550 --> 00:46:03,785
CALLED THE TRANSFRAME REGION.

1090
00:46:03,785 --> 00:46:09,657
IT'S IN THE REGION OF OVERLAP 

1091
00:46:09,657 --> 00:46:13,027
BETWEEN GAG AND POL AND IT'S 

1092
00:46:13,027 --> 00:46:15,830
REALLY RIGHT UPSTREAM OF THE 

1093
00:46:15,830 --> 00:46:16,231
PROTEASE.

1094
00:46:16,231 --> 00:46:17,966
THAT'S WHY IT'S CALLED 

1095
00:46:17,966 --> 00:46:18,867
TRANSFRAME REGION.

1096
00:46:18,867 --> 00:46:21,936
NOW, ONE SET OF MUTANTS, THE 

1097
00:46:21,936 --> 00:46:25,173
ONES, THE VARYING SET HAD 

1098
00:46:25,173 --> 00:46:28,443
SUBSTITUTED F, AND THEY IN FACT 

1099
00:46:28,443 --> 00:46:31,513
HAD NO CHANGE IN THE POL DOMAIN.

1100
00:46:31,513 --> 00:46:33,414
SO THAT WAS REALLY PERPLEXING 

1101
00:46:33,414 --> 00:46:36,184
BECAUSE THERE WAS NO OBVIOUS 

1102
00:46:36,184 --> 00:46:40,588
REASON WHY, THEN -- THERE SHOULD

1103
00:46:40,588 --> 00:46:41,856
HAVE BEEN MORE RESPONSE TO ART.

1104
00:46:41,856 --> 00:46:44,759
THE OTHER ONE GAVE US A REASON 

1105
00:46:44,759 --> 00:46:47,061
BECAUSE IF IN FACT THE MUTATION 

1106
00:46:47,061 --> 00:46:49,597
WAS ALANINE, THEN IN THE 

1107
00:46:49,597 --> 00:46:52,333
OVERLAPPING FRAME, IT ACTUALLY 

1108
00:46:52,333 --> 00:46:56,104
CHANGED THE RESIDUE TO A CYST 

1109
00:46:56,104 --> 00:46:57,272
TEEN AND THAT WAS HAPPENING 

1110
00:46:57,272 --> 00:46:59,240
RIGHT AT THE CLEAVAGE SITE AT 

1111
00:46:59,240 --> 00:47:01,876
THE END TERMINUS OF THE 

1112
00:47:01,876 --> 00:47:02,443
PROTEASE.

1113
00:47:02,443 --> 00:47:03,878
SO WHAT WAS HAPPENING WAS THOSE 

1114
00:47:03,878 --> 00:47:07,148
VARIANTS WERE NOT MAKING THE 99 

1115
00:47:07,148 --> 00:47:09,450
AMINO ACID PROTEASE.

1116
00:47:09,450 --> 00:47:10,952
THEY WERE MAKING PROTEASE 

1117
00:47:10,952 --> 00:47:11,452
EXTENSIONS, SO YOU CAN 

1118
00:47:11,452 --> 00:47:13,288
IMMEDIATELY SEE WHY THEY MIGHT 

1119
00:47:13,288 --> 00:47:16,758
NOT BE RESPONSIVE TO A DRUG THAT

1120
00:47:16,758 --> 00:47:18,526
IS HONED TO FIT INTO IN THE 

1121
00:47:18,526 --> 00:47:20,762
SUBSTRATE BINDING SITE IN 

1122
00:47:20,762 --> 00:47:25,700
PROTEASE BECAUSE IT'S GOT AN 

1123
00:47:25,700 --> 00:47:28,336
EXTENSION AT ITS END TERMINUS.

1124
00:47:28,336 --> 00:47:29,871
SO WE WERE PARTICULARLY 

1125
00:47:29,871 --> 00:47:31,706
FASCINATED WITH THE VARIANTS, 

1126
00:47:31,706 --> 00:47:32,974
SUBSTITUTED F IN THE GAG FRAME 

1127
00:47:32,974 --> 00:47:34,776
AND THERE WAS NO CHANGE IN THE 

1128
00:47:34,776 --> 00:47:35,310
FOL FRAME.

1129
00:47:35,310 --> 00:47:37,412
AND THERE -- IN THE POL FRAME.

1130
00:47:37,412 --> 00:47:38,780
SO THERE WERE SEVERAL THINGS 

1131
00:47:38,780 --> 00:47:40,415
THAT WERE VERY CURIOUS.

1132
00:47:40,415 --> 00:47:43,551
NUMBER ONE, THE MUTATION, THESE 

1133
00:47:43,551 --> 00:47:46,421
VARIANTS WERE TEN TIMES MORE 

1134
00:47:46,421 --> 00:47:51,059
PREVALENT IN WIHS VARIANTS THAN 

1135
00:47:51,059 --> 00:47:57,665
IN THE LASS ALLAH -- LOSS ALLAH 

1136
00:47:57,665 --> 00:47:59,033
MONTHS DATABASE AND AT THE TIME,

1137
00:47:59,033 --> 00:48:05,773
I WAS TOLD THAT THE LOS ALAMOS 

1138
00:48:05,773 --> 00:48:07,075
DATABASE WAS ACTUALLY ARCHIVING 

1139
00:48:07,075 --> 00:48:08,543
MAINLY VARIANTS THAT HAD BEEN 

1140
00:48:08,543 --> 00:48:10,612
ISOLATED FROM HIV-INFECTED 

1141
00:48:10,612 --> 00:48:11,312
MALES, ALL RIGHT?

1142
00:48:11,312 --> 00:48:14,782
SO THAT WAS SORT OF INTRIGUING.

1143
00:48:14,782 --> 00:48:18,620
AND THEN, THE SECOND THING THAT 

1144
00:48:18,620 --> 00:48:22,457
WAS UNSUSPECTED -- SO I WAS 

1145
00:48:22,457 --> 00:48:23,424
COLLABORATING WITH ONE SITE.

1146
00:48:23,424 --> 00:48:25,793
THERE WERE MAYBE ABOUT A DOZEN 

1147
00:48:25,793 --> 00:48:27,662
SITES OF WIHS AROUND THE COUNTRY

1148
00:48:27,662 --> 00:48:29,297
AND BECAUSE I WAS COLLABORATING 

1149
00:48:29,297 --> 00:48:30,698
WITH ONE SITE IN THE BRONX, THE 

1150
00:48:30,698 --> 00:48:33,668
THOUGHT WAS, WELL, MAYBE THESE 

1151
00:48:33,668 --> 00:48:36,671
PARTICIPANTS ARE SOMEHOW, SOME 

1152
00:48:36,671 --> 00:48:38,339
WAY, IN CONTACT AND THEY SPREAD 

1153
00:48:38,339 --> 00:48:47,482
THE VIRUS FROM ONE TO THE OTHER.

1154
00:48:47,482 --> 00:48:52,086
SO THE LAS ALAMOS COLLABORATOR 

1155
00:48:52,086 --> 00:48:53,288
PREPARED THIS TREE AND SHOWED US

1156
00:48:53,288 --> 00:48:55,323
THAT, NO, THESE VARIANTS 

1157
00:48:55,323 --> 00:48:56,591
APPARENTLY AROSE INDEPENDENTLY 

1158
00:48:56,591 --> 00:48:57,659
IN THE PARTICIPANTS AND THEY 

1159
00:48:57,659 --> 00:49:01,129
WERE NOT RELATED.

1160
00:49:01,129 --> 00:49:04,332
SO THAT WAS FASCINATING.

1161
00:49:04,332 --> 00:49:08,636
WE COULD RECAPITULATE THAT THE 

1162
00:49:08,636 --> 00:49:12,407
MUTATIONS CONFERRED INDON'T 

1163
00:49:12,407 --> 00:49:14,409
INAVIR RESISTANCE TO THE CONTEXT

1164
00:49:14,409 --> 00:49:17,979
OF NL4-3 LABORATORY STRAIN, AND 

1165
00:49:17,979 --> 00:49:21,382
YOU HAVE TO REMEMBER THAT THE 

1166
00:49:21,382 --> 00:49:24,452
INDINAVIR, THE REASON WE CHOSE 

1167
00:49:24,452 --> 00:49:27,522
IT WAS PART OF THE ART, AND SO 

1168
00:49:27,522 --> 00:49:30,325
WE COULD RECAPITULATE IN THAT IN

1169
00:49:30,325 --> 00:49:32,560
THE CONTEXT OF NL4-3.

1170
00:49:32,560 --> 00:49:35,463
THE MUTANTS WERE INDEED 

1171
00:49:35,463 --> 00:49:35,730
SENSITIVE.

1172
00:49:35,730 --> 00:49:37,198
SO IF THERE'S NO DRUG, THEN ONE 

1173
00:49:37,198 --> 00:49:44,772
OF THE MUTANTS, THE S40A WAS AS 

1174
00:49:44,772 --> 00:49:45,973
FIT AS THE WILD TYPE.

1175
00:49:45,973 --> 00:49:47,809
THE OTHER ONE WAS LESS FIT.

1176
00:49:47,809 --> 00:49:49,277
SO IT CASS CARRYING THIS 

1177
00:49:49,277 --> 00:49:52,113
MUTATION AT A COST, BUT IN THE 

1178
00:49:52,113 --> 00:49:56,351
PRESENCE OF INDINAVIR, THEN, IN 

1179
00:49:56,351 --> 00:49:58,619
FACT, THE S40A, HERE'S THE WILD 

1180
00:49:58,619 --> 00:50:01,622
TYPE DOWN HERE, IT WAS MORE FIT 

1181
00:50:01,622 --> 00:50:05,827
THAN THE WILD TYPE AND THE S40F 

1182
00:50:05,827 --> 00:50:08,496
AT LEAST BECAME AS FIT AS THE 

1183
00:50:08,496 --> 00:50:09,230
WILD TYPE.

1184
00:50:09,230 --> 00:50:13,334
SO IN BOTH CASES, THE MUTANTS 

1185
00:50:13,334 --> 00:50:17,038
WERE DOING BETTER RECAPITULATING

1186
00:50:17,038 --> 00:50:19,140
WHAT WAS HAPPENING IN VIVO.

1187
00:50:19,140 --> 00:50:21,776
I'M LOOKING FOR TIME.

1188
00:50:21,776 --> 00:50:23,778
ALL RIGHT.

1189
00:50:23,778 --> 00:50:26,514
NOW, ONCE AGAIN, WE BUILT THESE 

1190
00:50:26,514 --> 00:50:29,951
MUTATIONS INTO THE CONTEXT OF 

1191
00:50:29,951 --> 00:50:32,887
NL4-3, AND WE THEN ASKED WHAT 

1192
00:50:32,887 --> 00:50:33,654
HAPPENED.

1193
00:50:33,654 --> 00:50:35,189
AND SO OBVIOUSLY ON THE LEFT 

1194
00:50:35,189 --> 00:50:37,291
PANEL HERE, YOU CAN SEE IT.

1195
00:50:37,291 --> 00:50:41,295
IF IT'S WILD-TIME NL4-3, IT 

1196
00:50:41,295 --> 00:50:43,030
MAKES VIRAL PARTICLES THAT LOOK 

1197
00:50:43,030 --> 00:50:45,266
JUST LIKE NORMAL VIRAL PARTICLES

1198
00:50:45,266 --> 00:50:45,433
DO.

1199
00:50:45,433 --> 00:50:50,271
IF IN FACT THE MUTATION WAS 

1200
00:50:50,271 --> 00:50:51,973
S40A, IT COULD RELEASE VIRAL 

1201
00:50:51,973 --> 00:50:54,509
PARTICLES AS EFFICIENTLY AS THE 

1202
00:50:54,509 --> 00:50:55,243
WILD-TYPE, BUT THEY WERE NOT 

1203
00:50:55,243 --> 00:50:57,712
MATURE.

1204
00:50:57,712 --> 00:51:02,617
IF IN FACT IT WAS THE S40F 

1205
00:51:02,617 --> 00:51:06,120
VARIANT, THEN IT DIDN'T MAKE 

1206
00:51:06,120 --> 00:51:06,821
MATURE PARTICLES.

1207
00:51:06,821 --> 00:51:10,858
IT MADE IMMATURE PARTICLES, BUT 

1208
00:51:10,858 --> 00:51:12,894
THEY WERE ACTUALLY NOT RELEASED.

1209
00:51:12,894 --> 00:51:14,929
THEY WERE IN THE STRUCTURES 

1210
00:51:14,929 --> 00:51:18,599
WHICH RESEMBLES FILOPODIA.

1211
00:51:18,599 --> 00:51:21,035
SO THEN WE CAME TO UNDERSTAND 

1212
00:51:21,035 --> 00:51:23,404
THAT THAT COULD PROVIDE AN 

1213
00:51:23,404 --> 00:51:26,441
EXPLANATION FOR WHY THEY WOULD 

1214
00:51:26,441 --> 00:51:30,812
ESCAPE IF THEY'RE IN FILOPODIA, 

1215
00:51:30,812 --> 00:51:32,980
THEN THEY'RE NOT GOING TO BE 

1216
00:51:32,980 --> 00:51:35,249
SUBJECT TO DRUGS OR EVEN 

1217
00:51:35,249 --> 00:51:37,285
ANTIBODIES THAT MIGHT CLEAR.

1218
00:51:37,285 --> 00:51:39,320
AND ONCE AGAIN, GOING BACK TO 

1219
00:51:39,320 --> 00:51:40,488
THIS ISSUE OF WHETHER YOU HAVE 

1220
00:51:40,488 --> 00:51:44,725
LATE DOMAIN 1, 2, AND 3, FOR 

1221
00:51:44,725 --> 00:51:46,561
REASONS OF REDUNDANCY, WELL, 

1222
00:51:46,561 --> 00:51:49,030
YES, BUT I THINK THAT HERE'S 

1223
00:51:49,030 --> 00:51:51,299
ANOTHER REASON WHY YOU HAVE LATE

1224
00:51:51,299 --> 00:51:54,902
DOMAIN REDUNDANCY BECAUSE IT 

1225
00:51:54,902 --> 00:51:58,940
PERMITS THE VIRUS TO HIDE UNDER 

1226
00:51:58,940 --> 00:51:59,607
CIRCUMSTANCES WHERE THAT MIGHT 

1227
00:51:59,607 --> 00:52:02,276
BE PRACTICAL.

1228
00:52:02,276 --> 00:52:02,910
OKAY.

1229
00:52:02,910 --> 00:52:05,580
SO ONE QUESTION THAT WE ASKED 

1230
00:52:05,580 --> 00:52:07,915
AND WE'RE STILL ASKING NOW IS 

1231
00:52:07,915 --> 00:52:11,886
WHY WAS S40F TEN TIMES MORE 

1232
00:52:11,886 --> 00:52:15,289
PREVALENT IN THE WIHS VARIANTS 

1233
00:52:15,289 --> 00:52:20,862
THAN IN THE LAS ALAMOS DATABASE,

1234
00:52:20,862 --> 00:52:23,498
AND SO WHAT WE DID WAS IN 

1235
00:52:23,498 --> 00:52:25,600
COLLABORATION WITH TOM HOGUE, WE

1236
00:52:25,600 --> 00:52:28,269
GOT A NRMENT FOR MY GRANT FROM 

1237
00:52:28,269 --> 00:52:32,240
THE OFFICE OF WOMEN'S STUDIES AT

1238
00:52:32,240 --> 00:52:34,408
NIH, AND WHAT WE HAVE BEEN 

1239
00:52:34,408 --> 00:52:39,914
TRYING TO DO IS TO LOOK AT HOW 

1240
00:52:39,914 --> 00:52:41,883
THE MUTANTS FAIRE IN DIFFERENT 

1241
00:52:41,883 --> 00:52:44,485
KINDS OF TISSUES WHEN THEY ARE 

1242
00:52:44,485 --> 00:52:46,420
DERIVED FROM MALES OR FEMALES.

1243
00:52:46,420 --> 00:52:48,489
SO THE STORY IS NOT YET DONE, 

1244
00:52:48,489 --> 00:52:50,491
AND SO THAT'S WHY I'M NOT 

1245
00:52:50,491 --> 00:52:52,560
SHARING IT WITH YOU TODAY.

1246
00:52:52,560 --> 00:52:57,665
BUT I THINK IT WOULD BE REALLY A

1247
00:52:57,665 --> 00:52:59,033
SOURCE WHERE I KNOW OF WHERE 

1248
00:52:59,033 --> 00:53:01,903
THERE'S SOME BIAS RELATING TO 

1249
00:53:01,903 --> 00:53:02,970
THE KIND OF VARIANTS THAT WE 

1250
00:53:02,970 --> 00:53:03,905
SAW.

1251
00:53:03,905 --> 00:53:06,974
ALL RIGHT, BACK TO THE TSG.

1252
00:53:06,974 --> 00:53:10,411
SO AT CERTAIN POINTS, WE DECIDED

1253
00:53:10,411 --> 00:53:13,814
THAT WE SHOULD TRY TO USE WHAT 

1254
00:53:13,814 --> 00:53:16,851
WE KNEW ABOUT TSG101 AND NEDD4 

1255
00:53:16,851 --> 00:53:20,054
TO SEE IF WE COULD TRANSLATE ANY

1256
00:53:20,054 --> 00:53:22,790
OF THIS INFORMATION INTO 

1257
00:53:22,790 --> 00:53:25,693
SOMETHING THAT MIGHT BE A 

1258
00:53:25,693 --> 00:53:27,128
POTENTIAL ANTIVIRAL.

1259
00:53:27,128 --> 00:53:30,164
AND THAT WAS WHEN, ONCE AGAIN, 

1260
00:53:30,164 --> 00:53:32,733
WE WENT TO A LIBRARY AGAIN.

1261
00:53:32,733 --> 00:53:36,971
THIS TIME IT WAS A PROPRIETARY 

1262
00:53:36,971 --> 00:53:42,176
LIBRARY OF SMALL MOLECULES, AND 

1263
00:53:42,176 --> 00:53:47,615
WE FED IT THE RECOMBINANT UEV 

1264
00:53:47,615 --> 00:53:52,420
DOMAIN, AND FED IT THE 

1265
00:53:52,420 --> 00:53:55,423
RECOMBINANT NEDD4, AND OUT OF 

1266
00:53:55,423 --> 00:53:56,857
THAT, WE PULLED OUT ABOUT A HALF

1267
00:53:56,857 --> 00:53:58,993
A DOZEN HITS.

1268
00:53:58,993 --> 00:54:01,062
THIS WAS ABOUT 80,000 COMPOUNDS 

1269
00:54:01,062 --> 00:54:03,431
THAT WERE SCREENED AND WE PULLED

1270
00:54:03,431 --> 00:54:05,600
OUT ABOUT A HALF DOZEN HITS, AND

1271
00:54:05,600 --> 00:54:07,101
THEN WENT TO WORK TRYING TO 

1272
00:54:07,101 --> 00:54:09,637
VALIDATE THOSE HITS.

1273
00:54:09,637 --> 00:54:12,807
AND OUT OF THAT VALIDATION CAME 

1274
00:54:12,807 --> 00:54:13,674
THE PRAZOLES.

1275
00:54:13,674 --> 00:54:17,678
NOW, PRAZOLES, MANY OF YOU MAY 

1276
00:54:17,678 --> 00:54:21,682
KNOW, THEY ARE FDA-APPROVED 

1277
00:54:21,682 --> 00:54:25,152
COMPOUNDS FOR GASTROINTESTINAL 

1278
00:54:25,152 --> 00:54:28,723
DISEASE, AND WHAT THEY DO IN 

1279
00:54:28,723 --> 00:54:33,094
PARTICULAR IS THEY BIND TO AN 

1280
00:54:33,094 --> 00:54:35,930
ATP-ASE, WHICH IS IN THE SMALL 

1281
00:54:35,930 --> 00:54:38,199
INTESTINE AREA, AND THEY 

1282
00:54:38,199 --> 00:54:41,268
INTERFERE WITH ITS ABILITY TO 

1283
00:54:41,268 --> 00:54:44,672
PUMP ACID -- I'M SORRY -- YEAH, 

1284
00:54:44,672 --> 00:54:47,608
I'M SORRY, THEY INTERFERE WITH 

1285
00:54:47,608 --> 00:54:49,143
ITS ABILITY TO PUMP ACID SO 

1286
00:54:49,143 --> 00:54:52,813
THAT, IN FACT, ACID DOESN'T 

1287
00:54:52,813 --> 00:54:57,652
BUILD UP IN THE INTESTINE.

1288
00:54:57,652 --> 00:55:01,355
AND SO THE THING IS THAT IN 

1289
00:55:01,355 --> 00:55:04,125
USING IT AS AN ANTIVIRAL, IT 

1290
00:55:04,125 --> 00:55:05,793
DOESN'T ACTUALLY WORK THE SAME 

1291
00:55:05,793 --> 00:55:06,260
WAY.

1292
00:55:06,260 --> 00:55:07,528
THAT IS TO SAY, IT HAS TO BE 

1293
00:55:07,528 --> 00:55:08,729
TAKEN UP INTO THE CELL.

1294
00:55:08,729 --> 00:55:12,600
IF YOU PREVENT THEM FROM GOING 

1295
00:55:12,600 --> 00:55:14,435
INTO THE CELL, THEY CAN FUNCTION

1296
00:55:14,435 --> 00:55:16,203
THE WAY THEY DO IN THE BODY, BUT

1297
00:55:16,203 --> 00:55:20,474
THEY WON'T INHIBIT THE VIRUS.

1298
00:55:20,474 --> 00:55:23,044
SO IT TURNS OUT THEN THAT THEIR 

1299
00:55:23,044 --> 00:55:25,746
USE IN THIS WAY IS A -- WHAT 

1300
00:55:25,746 --> 00:55:29,050
THEY CALL A COUNTER INDICATION 

1301
00:55:29,050 --> 00:55:30,384
OR ANOTHER INDICATION OF THIS 

1302
00:55:30,384 --> 00:55:30,651
COMPOUND.

1303
00:55:30,651 --> 00:55:33,187
AND SO WE BEGAN TO STUDY THEM 

1304
00:55:33,187 --> 00:55:35,356
AND WHAT THEY MIGHT ACTUALLY 

1305
00:55:35,356 --> 00:55:39,060
REVEAL TO US ABOUT WHAT'S GOING 

1306
00:55:39,060 --> 00:55:42,196
ON IN TERMS OF TSG101, NEDD4, 

1307
00:55:42,196 --> 00:55:47,134
HIV ASSEMBLY AND BUDDING.

1308
00:55:47,134 --> 00:55:48,202
SO OUR COLLABORATORS WERE ABLE 

1309
00:55:48,202 --> 00:55:50,471
TO IDENTIFY THE SITE IN THE UEV 

1310
00:55:50,471 --> 00:55:53,407
DOMAIN OF TSG WHERE THE PRAZOLES

1311
00:55:53,407 --> 00:55:56,510
DOCK AND IT TURNS OUT THAT THEY 

1312
00:55:56,510 --> 00:55:59,346
DOCK ON ONE OF THE 

1313
00:55:59,346 --> 00:56:00,047
UBIQUITIN-BINDING REGIONS IN THE

1314
00:56:00,047 --> 00:56:01,048
TSG MOLECULE.

1315
00:56:01,048 --> 00:56:04,585
AND WHEN THEY DOCK, THEY PREVENT

1316
00:56:04,585 --> 00:56:05,453
UB FROM BINDING.

1317
00:56:05,453 --> 00:56:08,956
YOU CAN SEE IN FAMILIAR A HERE, 

1318
00:56:08,956 --> 00:56:10,891
I HAVE UBIQUITIN DOCKED IN TO 

1319
00:56:10,891 --> 00:56:12,593
THE UEV.

1320
00:56:12,593 --> 00:56:15,896
IT CONTACTS UEV IN A NUMBER OF 

1321
00:56:15,896 --> 00:56:17,364
POINTS, BUT IF THE PRAZOLES 

1322
00:56:17,364 --> 00:56:19,133
BINDING THERE, IT DISPLACES THE 

1323
00:56:19,133 --> 00:56:22,103
UEV AND PREVENTS THAT.

1324
00:56:22,103 --> 00:56:23,437
THE PRAZOLES, ONE OF THE THINGS 

1325
00:56:23,437 --> 00:56:25,139
I MENTIONED TO YOU BEFORE IS 

1326
00:56:25,139 --> 00:56:27,775
THAT TSG BY INTERACTING WITH UEV

1327
00:56:27,775 --> 00:56:30,911
AND INTERFERING WITH HOW IT 

1328
00:56:30,911 --> 00:56:34,348
HANDLES UBIQUITIN, IT THEN IS 

1329
00:56:34,348 --> 00:56:36,984
ABLE TO STABILIZE NEDD4 SO IT 

1330
00:56:36,984 --> 00:56:39,019
DOESN'T TURN OVER AS RAPIDLY, 

1331
00:56:39,019 --> 00:56:42,790
AND THAT'S WHAT YOU SEE HERE, 

1332
00:56:42,790 --> 00:56:44,859
THAT IF IN FACT YOU HAVE GOT AN 

1333
00:56:44,859 --> 00:56:46,594
INCREASING AMOUNT OF A PRAZOLE, 

1334
00:56:46,594 --> 00:56:48,295
THEN YOU WILL, INSTEAD OF 

1335
00:56:48,295 --> 00:56:53,434
PROTECTING NEDD4, THE NEDD4 IS 

1336
00:56:53,434 --> 00:56:55,970
NO LONGER PROTECTED BY TSG101, 

1337
00:56:55,970 --> 00:56:57,171
YOU LOSE IT.

1338
00:56:57,171 --> 00:56:59,006
IF IN FACT YOU USE A TSG101 

1339
00:56:59,006 --> 00:57:03,410
MUTANT THAT CAN'T BIND THE 

1340
00:57:03,410 --> 00:57:04,411
PRAZOLES, THEN YOU CAN'T 

1341
00:57:04,411 --> 00:57:06,480
MAINTAIN PROTECTION OF NEDD4 

1342
00:57:06,480 --> 00:57:06,947
THROUGH THAT.

1343
00:57:06,947 --> 00:57:10,451
SO THAT MEANS THAT THE PRAZOLE, 

1344
00:57:10,451 --> 00:57:12,753
BY BLOCKING TSG'S ABILITY TO 

1345
00:57:12,753 --> 00:57:17,424
BIND UB, ARE ACTUALLY ABLE TO 

1346
00:57:17,424 --> 00:57:24,598
DISABLE NEDD4 IN ADDITION.

1347
00:57:24,598 --> 00:57:26,500
AND YOU CAN INTERFERE WITH 

1348
00:57:26,500 --> 00:57:27,101
EITHER SIDE.

1349
00:57:27,101 --> 00:57:28,536
YOU CAN INTERFERE WITH THE TSG 

1350
00:57:28,536 --> 00:57:31,438
SIDE WITH A SMALL MOLECULE OR 

1351
00:57:31,438 --> 00:57:34,508
YOU CAN INTERFERE WITH THE NEDD4

1352
00:57:34,508 --> 00:57:36,243
SIDE WITH A SMALL MOLECULE, AND 

1353
00:57:36,243 --> 00:57:39,113
YOU'LL GET THE SAME RESULT, 

1354
00:57:39,113 --> 00:57:42,349
WHICH IS TWO ARREST BUDDING AT 

1355
00:57:42,349 --> 00:57:45,953
AN EARLY STAGE, EARLIER THAN THE

1356
00:57:45,953 --> 00:57:48,255
STAGE WHERE, FOR EXAMPLE, YOU 

1357
00:57:48,255 --> 00:57:51,292
GET THE TETHERED NET, WHICH IS 

1358
00:57:51,292 --> 00:57:55,362
THE CLASSICAL FORM OF BUDDING 

1359
00:57:55,362 --> 00:57:55,596
ARREST.

1360
00:57:55,596 --> 00:57:58,732
HERE, THE WHOLE PARTICLE IS 

1361
00:57:58,732 --> 00:57:59,600
MADE.

1362
00:57:59,600 --> 00:58:03,604
HERE, IT'S ARRESTED EARLIER.

1363
00:58:03,604 --> 00:58:06,607
WE'RE FINDING -- SO AT THIS 

1364
00:58:06,607 --> 00:58:08,242
POINT, WE'VE MADE LOTS OF 

1365
00:58:08,242 --> 00:58:11,178
PRAZOLE DERIVATIVES IN 

1366
00:58:11,178 --> 00:58:13,514
COLLABORATION WITH OUR MEDICINAL

1367
00:58:13,514 --> 00:58:15,816
CHEMIST FRIEND, AND JUST LIKE 

1368
00:58:15,816 --> 00:58:17,618
THE COMMERCIAL PRAZOLES, WHICH 

1369
00:58:17,618 --> 00:58:19,887
AS YOU KNOW OR MAY KNOW COMES IN

1370
00:58:19,887 --> 00:58:22,122
ALL KINDS OF FLAVORS, SO YOU CAN

1371
00:58:22,122 --> 00:58:24,725
TAKE A PRAZOLE FOR IMMEDIATE 

1372
00:58:24,725 --> 00:58:24,959
ACTION.

1373
00:58:24,959 --> 00:58:26,727
YOU CAN TAKE IT FOR OVERNIGHT 

1374
00:58:26,727 --> 00:58:26,961
ACTION.

1375
00:58:26,961 --> 00:58:27,962
YOU CAN TAKE IT FOR SIX-HOUR 

1376
00:58:27,962 --> 00:58:30,598
ACTION.

1377
00:58:30,598 --> 00:58:31,665
JUST LIKE THE COMMERCIAL 

1378
00:58:31,665 --> 00:58:35,469
PRAZOLES, WHEN WE LOOK AT OUR 

1379
00:58:35,469 --> 00:58:36,403
PRAZOLES, THEN THEY MAKE 

1380
00:58:36,403 --> 00:58:41,242
DISTINCT FOOTPRINTS ON THE TSG 

1381
00:58:41,242 --> 00:58:41,775
UEV MOLECULE.

1382
00:58:41,775 --> 00:58:43,444
THEY ARE ALL PRO DRUGS.

1383
00:58:43,444 --> 00:58:48,782
THEY ALL ATTACK A CYSTINE IN THE

1384
00:58:48,782 --> 00:58:51,018
TSG MOLECULE, BUT THEY DON'T 

1385
00:58:51,018 --> 00:58:52,586
NECESSARILY, JUST BECAUSE OF 

1386
00:58:52,586 --> 00:58:54,288
THEIR STRUCTURE, STICK IN THE 

1387
00:58:54,288 --> 00:58:55,189
SAME WAY.

1388
00:58:55,189 --> 00:58:59,593
THIS GAVE US THE IDEA THAT MAYBE

1389
00:58:59,593 --> 00:59:01,328
YOU COULD HAVE PRAZOLE VIRUS 

1390
00:59:01,328 --> 00:59:04,832
SPECIFIC TARGETING, AND WE'RE 

1391
00:59:04,832 --> 00:59:07,201
ENCOURAGED IN THAT THOUGHT BY 

1392
00:59:07,201 --> 00:59:10,471
LOOKING AT PRAZOLES DERIVATIVE 

1393
00:59:10,471 --> 00:59:12,172
EFFICACY WITH SARS.

1394
00:59:12,172 --> 00:59:15,209
SO IF YOU COMPARE HIV-1 AND 

1395
00:59:15,209 --> 00:59:19,346
SARS, WHAT YOU SEE IS THAT HERE 

1396
00:59:19,346 --> 00:59:21,315
IS A GUY WHO IS NOT -- FIRST OF 

1397
00:59:21,315 --> 00:59:24,451
ALL, JUST TAKE A COMMERCIAL 

1398
00:59:24,451 --> 00:59:24,685
PRAZOLE.

1399
00:59:24,685 --> 00:59:27,354
COMMERCIAL PRAZOLE, IT'S ONE OF 

1400
00:59:27,354 --> 00:59:29,790
THEM IS NOT PARTICULARLY 

1401
00:59:29,790 --> 00:59:31,358
EFFICACIOUS WITH EITHER BUD.

1402
00:59:31,358 --> 00:59:32,493
HERE'S ONE THAT IS NOT 

1403
00:59:32,493 --> 00:59:35,863
PARTICULARLY SELECTIVE AS FAR AS

1404
00:59:35,863 --> 00:59:37,831
HIV IS CONCERNED, BUT WITH SARS,

1405
00:59:37,831 --> 00:59:42,636
YOU KNOW, FOR US AT OUR STAGE OF

1406
00:59:42,636 --> 00:59:43,537
INVESTIGATION, WE'RE GETTING 

1407
00:59:43,537 --> 00:59:46,206
PRETTY GOOD SELECTIVITY WITH 

1408
00:59:46,206 --> 00:59:46,473
SARS.

1409
00:59:46,473 --> 00:59:47,841
ON THE OTHER HAND, TAKE THE GUY 

1410
00:59:47,841 --> 00:59:49,910
WHO IS NOT PARTICULAR SELECTIVE 

1411
00:59:49,910 --> 00:59:52,012
WITH RESPECT TO SARS, AND HE'S 

1412
00:59:52,012 --> 00:59:55,215
NOT BAD WITH RESPECT TO HIV.

1413
00:59:55,215 --> 00:59:58,385
SO THAT IS GIVING US THE 

1414
00:59:58,385 --> 01:00:03,524
IMPRESSION THAT MAYBE THE -- 

1415
01:00:03,524 --> 01:00:05,292
THESE COULD BE VERY FORMATIVE IN

1416
01:00:05,292 --> 01:00:06,894
TERMS OF HOW THEY -- INFORMATIVE

1417
01:00:06,894 --> 01:00:08,862
IN TERMS OF HOW THEY TELL US 

1418
01:00:08,862 --> 01:00:10,431
WHAT'S HAPPENING IN TRAFFIC WITH

1419
01:00:10,431 --> 01:00:11,832
THESE VARIOUS VIRUSES.

1420
01:00:11,832 --> 01:00:13,233
I DIDN'T INCLUDE A SLIDE TO SHOW

1421
01:00:13,233 --> 01:00:15,669
YOU HERE, BUT WE'VE ALSO DONE 

1422
01:00:15,669 --> 01:00:17,371
THIS SORT OF EXPERIMENT USING 

1423
01:00:17,371 --> 01:00:19,606
DIFFERENT CELL MACHINERIES WHERE

1424
01:00:19,606 --> 01:00:21,742
TSG AND ESCRT ARE INVOLVED, AND 

1425
01:00:21,742 --> 01:00:25,012
AGAIN, WE SEE SELECTIVITY.

1426
01:00:25,012 --> 01:00:26,747
THERE ARE CERTAIN MACHINE RIS 

1427
01:00:26,747 --> 01:00:29,483
THAT ARE PARTICULARLY 

1428
01:00:29,483 --> 01:00:31,018
SUSCEPTIBLE TO PRAZOLES, BUT 

1429
01:00:31,018 --> 01:00:32,786
THERE ARE ONES THAT SEEM TO 

1430
01:00:32,786 --> 01:00:34,288
IGNORE IT ENTIRELY.

1431
01:00:34,288 --> 01:00:36,290
EVEN THOUGH THEY'RE ALL -- THEY 

1432
01:00:36,290 --> 01:00:39,326
ALL HAVE TSG INVOLVEMENT.

1433
01:00:39,326 --> 01:00:41,762
ALL RIGHT.

1434
01:00:41,762 --> 01:00:44,131
WELL, THERE'S A MODEL TO LEAVE 

1435
01:00:44,131 --> 01:00:45,766
YOU WITH.

1436
01:00:45,766 --> 01:00:47,201
ESSENTIALLY, WE'RE THINKING THAT

1437
01:00:47,201 --> 01:00:51,105
THE TSG101 UEV DOMAIN, ITS JOB 

1438
01:00:51,105 --> 01:00:57,945
IS TO ACTIVATE NEDD4 BY TAKING 

1439
01:00:57,945 --> 01:01:00,781
IT OUT OF ITS AUTO-INHIBITION 

1440
01:01:00,781 --> 01:01:01,882
STATE AT LEAST FOR THE PERIOD 

1441
01:01:01,882 --> 01:01:04,551
THAT THE VIRUS NEEDS IT, AND 

1442
01:01:04,551 --> 01:01:07,755
DOES SO BY ESSENTIALLY 

1443
01:01:07,755 --> 01:01:10,624
INTERFERING WITH THIS NATURAL C2

1444
01:01:10,624 --> 01:01:14,294
HECT INHIBITION THAT THE PROTEIN

1445
01:01:14,294 --> 01:01:15,963
WOULD BE FOUND UNDER IN THE 

1446
01:01:15,963 --> 01:01:16,864
CELL.

1447
01:01:16,864 --> 01:01:20,200
AND ONCE THAT IS THE CASE, THEN 

1448
01:01:20,200 --> 01:01:22,336
THE ENZYME CAN, YOU KNOW, BRING 

1449
01:01:22,336 --> 01:01:25,406
THE COHORTS THAT IT BRINGS.

1450
01:01:25,406 --> 01:01:26,573
IF IT'S IN THE CASE OF THE 

1451
01:01:26,573 --> 01:01:29,410
COHORT THAT CAN ENABLE US TO 

1452
01:01:29,410 --> 01:01:31,678
TAKE ADVANTAGE OF HIDING IN 

1453
01:01:31,678 --> 01:01:33,781
FILOPO DI-A, IT CAN BRING THAT, 

1454
01:01:33,781 --> 01:01:35,516
TOO, AND THEN YOU GET A 

1455
01:01:35,516 --> 01:01:38,952
SITUATION WHERE THE VIRUS CAN 

1456
01:01:38,952 --> 01:01:44,058
FIND SOME OTHER WAY OF 

1457
01:01:44,058 --> 01:01:44,658
MECHANISM.

1458
01:01:44,658 --> 01:01:46,060
SO WHOO HAVE I TOLD YOU?

1459
01:01:46,060 --> 01:01:47,828
I TOLD YOU VIRAL L DOMAINS ARE 

1460
01:01:47,828 --> 01:01:49,196
DOCKING SITES FOR THE 

1461
01:01:49,196 --> 01:01:50,964
RECRUITMENT OF CELLULAR FACTORS 

1462
01:01:50,964 --> 01:01:53,100
TSG101 AND NEDD4.

1463
01:01:53,100 --> 01:01:57,204
THE TSG101 UEV DOMAIN ACTIVATES 

1464
01:01:57,204 --> 01:02:00,107
NEDD4 BY INTERFERING WITH THE 

1465
01:02:00,107 --> 01:02:00,741
AUTO-INHIBITION MECHANISM BY 

1466
01:02:00,741 --> 01:02:04,678
WHICH THE CELL KEEPS THE ENZYME 

1467
01:02:04,678 --> 01:02:04,912
SILENT.

1468
01:02:04,912 --> 01:02:08,082
THE TSG REGULATES, THIS WE 

1469
01:02:08,082 --> 01:02:11,752
BELIEVE BY SUPPRESSING THE 

1470
01:02:11,752 --> 01:02:12,386
AUTO-INHIBITION THAT TYPICALLY 

1471
01:02:12,386 --> 01:02:14,621
IS THE WAY -- IS THE CELL'S WAY 

1472
01:02:14,621 --> 01:02:17,658
OF SORT OF TEMPORALLY 

1473
01:02:17,658 --> 01:02:18,625
CONTROLLING THE ENZYME'S 

1474
01:02:18,625 --> 01:02:18,926
ACTIVITY.

1475
01:02:18,926 --> 01:02:24,598
THE NEDD4 ACTIVATION PROMOTES UB

1476
01:02:24,598 --> 01:02:25,365
MODIFICATION EVENTS THAT 

1477
01:02:25,365 --> 01:02:28,602
FACILITATE THE BUDDING OF 

1478
01:02:28,602 --> 01:02:30,304
CELL-FREE VIRUS AND THE VIRUS AT

1479
01:02:30,304 --> 01:02:33,674
LEAST AS FAR AS PEOPLE LIVING 

1480
01:02:33,674 --> 01:02:36,076
WITH HIV TELL US, THE VIRUS CAN 

1481
01:02:36,076 --> 01:02:43,183
GET ITS WAY AROUND THAT TOO BY 

1482
01:02:43,183 --> 01:02:44,485
COURTING THIS LATE DOMAIN 

1483
01:02:44,485 --> 01:02:44,952
MECHANISM.

1484
01:02:44,952 --> 01:02:46,253
INTERFERENCE WITH THIS KIND OF 

1485
01:02:46,253 --> 01:02:48,522
SIGNALING IS, WE BELIEVE, AN 

1486
01:02:48,522 --> 01:02:50,824
UNAPPRECIATED MECHANISM OF ART 

1487
01:02:50,824 --> 01:02:54,161
EVASION IN PEOPLE LIVING WITH 

1488
01:02:54,161 --> 01:02:54,328
HIV.

1489
01:02:54,328 --> 01:02:56,563
AND WE THINK THAT THESE PRAZOLES

1490
01:02:56,563 --> 01:02:58,665
ARE FDA-APPROVED AS SMALL 

1491
01:02:58,665 --> 01:03:02,336
MOLECULES AND WE KNOW THAT THEY 

1492
01:03:02,336 --> 01:03:07,541
INTERFERE WITH TSG-NEDD4 

1493
01:03:07,541 --> 01:03:08,475
INTERACTION, THEY ARE PROBABLY 

1494
01:03:08,475 --> 01:03:09,643
GOOD MEANS FOR TRYING TO FIND A 

1495
01:03:09,643 --> 01:03:10,978
WAY TO MAKE THEM SPECIFIC.

1496
01:03:10,978 --> 01:03:14,081
OBVIOUSLY NOTHING THAT I SHOWED 

1497
01:03:14,081 --> 01:03:16,450
YOU IS WHAT OUR SHORT-TERM GOAL 

1498
01:03:16,450 --> 01:03:19,186
IS, THAT IS TO SAY, GET 

1499
01:03:19,186 --> 01:03:24,057
SOMETHING IN THE SUBNANO MOLAR 

1500
01:03:24,057 --> 01:03:24,992
AFFINITY, BUT WE'RE THINKING IF 

1501
01:03:24,992 --> 01:03:26,693
ONE CAN ACHIEVE THAT, YOU HAVE A

1502
01:03:26,693 --> 01:03:28,395
SMALL MOLECULE THAT'S TARGETING 

1503
01:03:28,395 --> 01:03:32,199
HOST EXPLOITATION.

1504
01:03:32,199 --> 01:03:33,767
NOW, I HAVE TO THANK A WHOLE 

1505
01:03:33,767 --> 01:03:35,002
BUNCH OF PEOPLE.

1506
01:03:35,002 --> 01:03:35,936
I HAVE WONDERFUL -- I'VE ALWAYS 

1507
01:03:35,936 --> 01:03:37,204
HAD WONDERFUL, WONDERFUL 

1508
01:03:37,204 --> 01:03:39,072
COLLABORATORS AT STONY BROOK.

1509
01:03:39,072 --> 01:03:41,375
THESE ARE PEOPLE WHO WERE MOSTLY

1510
01:03:41,375 --> 01:03:43,377
INVOLVED IN THE WORK I SHOWED 

1511
01:03:43,377 --> 01:03:46,980
YOU TODAY, AND THEN AT MT. 

1512
01:03:46,980 --> 01:03:51,351
SINAI, BOTH VIVIANA AND ALSO BEN

1513
01:03:51,351 --> 01:03:54,054
CHEN, WITH WHOM WE'RE DOING 

1514
01:03:54,054 --> 01:03:56,657
CELL-CELL TRANSFER KINDS OF 

1515
01:03:56,657 --> 01:03:58,158
EXPERIMENTS, TOM AND THEN MY 

1516
01:03:58,158 --> 01:04:00,994
STRUCTURAL BIOLOGY COLLABORATORS

1517
01:04:00,994 --> 01:04:04,731
AT NIH AND MEDICINAL CHEMISTRY 

1518
01:04:04,731 --> 01:04:07,501
AS WELL AS OUR FRIENDS AT 

1519
01:04:07,501 --> 01:04:07,768
MOREHOUSE.

1520
01:04:07,768 --> 01:04:09,870
AND THEN ALSO OUR COMPUTATIONAL 

1521
01:04:09,870 --> 01:04:13,707
BIOLOGIST FRIENDS AT STONY BROOK

1522
01:04:13,707 --> 01:04:20,414
AND ALSO C CHOPEING CHEN.

1523
01:04:20,414 --> 01:04:21,215
SO THANK YOU VERY, VERY MUCH.

1524
01:04:21,215 --> 01:04:28,755
[ APPLAUSE ]

1525
01:04:28,755 --> 01:04:34,228
>> WE HAVE TIME FOR
QUESTIONS.

1526
01:04:34,228 --> 01:04:36,063
>> IN DESCRIBING FIRST OF
ALL, 

1527
01:04:36,063 --> 01:04:37,464
WHAT -- WHAT A DISCUSSION.

1528
01:04:37,464 --> 01:04:40,400
IN THE SLIDE WHERE YOU SHOWED 

1529
01:04:40,400 --> 01:04:41,969
THAT YOU TREAT INCREASING 

1530
01:04:41,969 --> 01:04:43,737
AMOUNTS OF THE PRAZOLES, I 

1531
01:04:43,737 --> 01:04:45,405
FORGOT WHAT IT WAS CALLED, AND 

1532
01:04:45,405 --> 01:04:50,244
YOU'RE ALSO ADDING NEDD4-2, I 

1533
01:04:50,244 --> 01:04:52,145
GUESS I'M NOT SURE, SINCE YOU'RE

1534
01:04:52,145 --> 01:04:54,815
ADDING NEDD4-2, THAT'S WHY YOU 

1535
01:04:54,815 --> 01:04:56,883
CAN SEE IT, BUT WOULD THAT SAME 

1536
01:04:56,883 --> 01:04:58,752
HAVE A SIMILAR EFFECT ON THE 

1537
01:04:58,752 --> 01:05:02,022
OTHER FORM OF NEDD4?

1538
01:05:02,022 --> 01:05:03,690
>> SO THAT'S VERY
INTERESTING.

1539
01:05:03,690 --> 01:05:05,726
SO I THINK THAT -- SO THE 

1540
01:05:05,726 --> 01:05:07,327
QUESTION WAS IN THE EXPERIMENT 

1541
01:05:07,327 --> 01:05:10,097
WHERE WE WANTED TO ASK WHETHER 

1542
01:05:10,097 --> 01:05:12,266
THE PRAZOLES, BECAUSE THEY'RE 

1543
01:05:12,266 --> 01:05:15,736
INTERACTING WITH TSG, WOULD THEY

1544
01:05:15,736 --> 01:05:18,238
IN FACT IMPACT A NEDD4 

1545
01:05:18,238 --> 01:05:20,073
DETERMINED FUNCTION, RIGHT?

1546
01:05:20,073 --> 01:05:21,975
SO THE THING IS THAT WHAT'S VERY

1547
01:05:21,975 --> 01:05:25,045
IMPORTANT FOR THE NEDD4 

1548
01:05:25,045 --> 01:05:26,280
DETERMINED FUNCTION IS THE 

1549
01:05:26,280 --> 01:05:29,683
UBIQUITIN MOLECULES THAT TSG IS 

1550
01:05:29,683 --> 01:05:29,916
HOLDING.

1551
01:05:29,916 --> 01:05:32,586
AND SO THE REASON THAT THE 

1552
01:05:32,586 --> 01:05:34,888
PRAZOLES WOULD BE EFFECTIVE IS 

1553
01:05:34,888 --> 01:05:36,957
BECAUSE, AS LONG AS IT'S 

1554
01:05:36,957 --> 01:05:39,993
INTERACTING WITH A NEDD4 THAT 

1555
01:05:39,993 --> 01:05:41,995
NEEDS THOSE UBs, THEN THE ANSWER

1556
01:05:41,995 --> 01:05:43,063
IS YES.

1557
01:05:43,063 --> 01:05:45,799
NOW, THE REASON THAT THE UBs ARE

1558
01:05:45,799 --> 01:05:48,735
SO IMPORTANT, IF YOU LOOK AT 

1559
01:05:48,735 --> 01:05:51,605
THE -- HOW WE MODEL THE 

1560
01:05:51,605 --> 01:05:54,741
STRUCTURE, I SAID BEFORE THAT 

1561
01:05:54,741 --> 01:05:58,312
THE TSG'S UEV DOMAIN, IT'S A 

1562
01:05:58,312 --> 01:05:59,513
UBIQUITIN E2 VARIANT.

1563
01:05:59,513 --> 01:06:04,284
IF YOU TAKE THE CLOSEST RELATED 

1564
01:06:04,284 --> 01:06:06,019
E2 AND YOU COMPARE HOW IT 

1565
01:06:06,019 --> 01:06:07,087
INTERACTS WITH UBIQUITIN TO HOW 

1566
01:06:07,087 --> 01:06:09,956
TSG INTERACTS WITH UBIQUITIN, IT

1567
01:06:09,956 --> 01:06:12,092
IS NOT THE SAME.

1568
01:06:12,092 --> 01:06:14,895
AND SO THE TSG UEV IS HOLDING 

1569
01:06:14,895 --> 01:06:17,831
THE UB IN A MANNER THAT WOULD 

1570
01:06:17,831 --> 01:06:21,635
APPEAR TO BE MODULATING HOW THE 

1571
01:06:21,635 --> 01:06:24,705
NEDD4 IS ABLE TO TRANSFER THE UB

1572
01:06:24,705 --> 01:06:26,740
TO SUBSTRATE AND TO ITSELF.

1573
01:06:26,740 --> 01:06:29,309
AND SO WE THINK THAT THAT'S AT 

1574
01:06:29,309 --> 01:06:32,346
THE CRUX OF HOW IT'S ABLE TO 

1575
01:06:32,346 --> 01:06:34,848
CONTROL WHAT NEDD4 DOES, AND AS 

1576
01:06:34,848 --> 01:06:37,250
LONG AS THE PRAZOLES ARE 

1577
01:06:37,250 --> 01:06:39,986
INTERFERING WITH THAT TSG 

1578
01:06:39,986 --> 01:06:44,858
FUNCTION, IT CAN HIT NEDD4-2. 

1579
01:06:44,858 --> 01:06:48,295
>> GREAT TALK.

1580
01:06:48,295 --> 01:06:53,200
SO HIV IS PRIMARY -- FIV, THEY 

1581
01:06:53,200 --> 01:06:55,902
RADIATED TO T CELLS FOR 

1582
01:06:55,902 --> 01:06:58,805
MACROPHAGE TROPISM.

1583
01:06:58,805 --> 01:07:01,108
WHEN ONE LOOKS AT LATE DOMAIN, 

1584
01:07:01,108 --> 01:07:05,011
ARE THERE LESS LATE DOMAINS IN, 

1585
01:07:05,011 --> 01:07:07,147
LIKE, THIS -- AND THE REDUNDANCY

1586
01:07:07,147 --> 01:07:10,250
THAT ONE SEES IN HIV WITH 

1587
01:07:10,250 --> 01:07:12,219
GREATER CELL PURPOSE?

1588
01:07:12,219 --> 01:07:14,287
>> LET ME SEE IF I REPEAT
THE 

1589
01:07:14,287 --> 01:07:15,489
QUESTION CORRECTLY, WHICH WAS 

1590
01:07:15,489 --> 01:07:19,226
THAT IF YOU TAKE SOME OF THE 

1591
01:07:19,226 --> 01:07:22,529
OTHER MEMBERS OF THE FAMILY, 

1592
01:07:22,529 --> 01:07:26,833
LET'S SAY WITHIN THE LATENCIES, 

1593
01:07:26,833 --> 01:07:28,001
NOT NECESSARILY OUTSIDE, BUT 

1594
01:07:28,001 --> 01:07:29,536
WITHIN, CAN YOU SEE A 

1595
01:07:29,536 --> 01:07:32,672
CORRELATION BETWEEN HOW 

1596
01:07:32,672 --> 01:07:34,274
EFFICIENTLY THEY ARE TRANSMITTED

1597
01:07:34,274 --> 01:07:38,345
AND LATE DOMAIN ACTIVITY.

1598
01:07:38,345 --> 01:07:40,947
>> DOES HIV, AS FAR AS YOU
KNOW,

1599
01:07:40,947 --> 01:07:42,716
DO THEY HAVE REDUNDANCY?

1600
01:07:42,716 --> 01:07:45,252
>> SO THEY DEFINITELY --
THE 

1601
01:07:45,252 --> 01:07:48,522
REASON I DON'T KNOW, BUT UEV WAS

1602
01:07:48,522 --> 01:07:49,990
A FAVORITE TO STUDY BECAUSE, AS 

1603
01:07:49,990 --> 01:07:52,726
I MENTIONED TO YOU, THERE'S 

1604
01:07:52,726 --> 01:07:57,431
DIFFERENT KINDS OF ALIX-BINDING 

1605
01:07:57,431 --> 01:07:57,864
MOT

1606
01:07:57,864 --> 01:07:58,865
MOTIFS, AND STRUCTURALLY THERE'S

1607
01:07:58,865 --> 01:08:01,234
SOME SIMILARITY, BUT IN TERMS OF

1608
01:08:01,234 --> 01:08:04,771
THE MOTIF ITSELF, THE SEQUENCE, 

1609
01:08:04,771 --> 01:08:05,906
THEY'RE NOT IDENTICAL.

1610
01:08:05,906 --> 01:08:08,208
SO EIV WOULD, AS FAR AS WE CAN 

1611
01:08:08,208 --> 01:08:10,177
SEE, BE VERY, VERY SIMILAR IN 

1612
01:08:10,177 --> 01:08:15,715
HOW IT INTERACTS WITH ALIX TO --

1613
01:08:15,715 --> 01:08:16,616
AS HIV.

1614
01:08:16,616 --> 01:08:17,884
WHAT WE DO NOT KNOW IS WHETHER 

1615
01:08:17,884 --> 01:08:20,187
OR NOT IN FACT IN THE CASE OF 

1616
01:08:20,187 --> 01:08:23,423
HIV-1, YOU CALL NEDD4-1.

1617
01:08:23,423 --> 01:08:26,193
IN THE CASE OF EIV, I DON'T KNOW

1618
01:08:26,193 --> 01:08:27,794
THAT YOU NECESSARILY CALL IT 

1619
01:08:27,794 --> 01:08:30,363
BECAUSE ALIX IS ITSELF ABLE TO 

1620
01:08:30,363 --> 01:08:33,433
HAVE MANY ADAPTOR FRIENDS THAT 

1621
01:08:33,433 --> 01:08:37,003
CAN CALL E3 LIGASE NROOP WHAT'S 

1622
01:08:37,003 --> 01:08:37,838
YOUR EXPLANATION FOR WHY THERE'S

1623
01:08:37,838 --> 01:08:38,872
SO MANY DOMAINS THEN?

1624
01:08:38,872 --> 01:08:41,308
>> I'M GOING TO TELL YOU
WHAT 

1625
01:08:41,308 --> 01:08:43,043
ALLEN SAID TO ME WHEN I WAS 

1626
01:08:43,043 --> 01:08:45,011
DEALING WITH HIM EARLIER TODAY.

1627
01:08:45,011 --> 01:08:47,414
HE IS SAID THAT WHENEVER ANYONE 

1628
01:08:47,414 --> 01:08:53,653
ASKS A QUESTION LIKE THAT, HE 

1629
01:08:53,653 --> 01:08:55,722
SAYS, WHY -- I'LL TAKE A STAB AT

1630
01:08:55,722 --> 01:09:01,194
ALLEN'S ANSWER.

1631
01:09:01,194 --> 01:09:08,401
>> IT'S RELATED SEQUENCES
WITHIN

1632
01:09:08,401 --> 01:09:11,505
THE TWO.

1633
01:09:11,505 --> 01:09:14,608
I WONDER WHAT COHORT IS ABLE THE

1634
01:09:14,608 --> 01:09:15,976
PATIENTS WHO ARE FEMALE.

1635
01:09:15,976 --> 01:09:19,546
AND MALE INDEX HOW THE 

1636
01:09:19,546 --> 01:09:22,449
FREQUENCE -- 

1637
01:09:22,449 --> 01:09:24,417
>> SO THE PERSON WHO 

1638
01:09:24,417 --> 01:09:25,952
COLLABORATED IN THIS WITH US, 

1639
01:09:25,952 --> 01:09:27,320
THE ANSWER IS -- THE QUESTION 

1640
01:09:27,320 --> 01:09:32,192
WAS IN TERMS OF THE LASALAMOS 

1641
01:09:32,192 --> 01:09:35,095
DATABASE, WHICH I WAS TOLD WAS 

1642
01:09:35,095 --> 01:09:36,897
PREDOMINANTLY MEN, I CHECKED IT 

1643
01:09:36,897 --> 01:09:37,898
FOR ACTUALLY 2000 -- WHATEVER 

1644
01:09:37,898 --> 01:09:41,468
WAS THE LATEST, '23 OR '24, AND 

1645
01:09:41,468 --> 01:09:42,636
NOW I THINK THEY REPORT 

1646
01:09:42,636 --> 01:09:45,171
SOMETHING LIKE 20 OR 30% 

1647
01:09:45,171 --> 01:09:46,473
VARIANTS FROM WOMEN.

1648
01:09:46,473 --> 01:09:48,108
BUT AT THE TIME THAT THIS WAS 

1649
01:09:48,108 --> 01:09:50,644
DONE WITH THE WIHS, I WAS TOLD 

1650
01:09:50,644 --> 01:09:54,614
THAT IT WAS, LIKE, IN THE 90s 

1651
01:09:54,614 --> 01:09:56,583
MEN, AND VERY, VERY FEW WOMEN.

1652
01:09:56,583 --> 01:09:58,718
SO THE FACT THAT IT WAS TEN 

1653
01:09:58,718 --> 01:10:01,254
TIMES HIGHER IN THE WIHS SEEMS 

1654
01:10:01,254 --> 01:10:01,955
SIGNIFICANT OR AT LEAST 

1655
01:10:01,955 --> 01:10:03,990
NOTEWORTHY.

1656
01:10:03,990 --> 01:10:09,329
>> BUT IN THE LAS ALAMOS,
THEY 

1657
01:10:09,329 --> 01:10:11,231
SHOULD HAVE THE SEX -- FEMALE 

1658
01:10:11,231 --> 01:10:13,867
AND MALE INSIDE THAT -- 

1659
01:10:13,867 --> 01:10:15,101
>> SO YOU MEAN RIGHT NOW?

1660
01:10:15,101 --> 01:10:17,637
I HAVEN'T GONE BACK TO DO THAT, 

1661
01:10:17,637 --> 01:10:17,904
ACTUALLY.

1662
01:10:17,904 --> 01:10:19,272
THAT WOULD BE REALLY INTERESTING

1663
01:10:19,272 --> 01:10:20,574
TO DO, YEAH.

1664
01:10:20,574 --> 01:10:21,908
>> AND THEN A SECOND
QUESTION 

1665
01:10:21,908 --> 01:10:26,212
IS, THE MUTATION THAT YOU FOUND 

1666
01:10:26,212 --> 01:10:29,482
S40, YOU THINK IF YOU CHANGE THE

1667
01:10:29,482 --> 01:10:30,116
BINDING, THEY PROJECT -- 

1668
01:10:30,116 --> 01:10:33,086
>> ABSOLUTELY.

1669
01:10:33,086 --> 01:10:34,254
THAT'S EXACTLY WHAT IT DOES.

1670
01:10:34,254 --> 01:10:37,657
SO IT'S -- I WON'T SAY 

1671
01:10:37,657 --> 01:10:39,459
COMPLICATED, BUT IT'S 

1672
01:10:39,459 --> 01:10:40,026
MULTI-STEPPED, BUT THAT'S 

1673
01:10:40,026 --> 01:10:43,597
EXACTLY WHAT IT DOES.

1674
01:10:43,597 --> 01:10:45,932
THANKS.

1675
01:10:45,932 --> 01:10:47,033
>> DO YOU KNOW WITHIN THE 

1676
01:10:47,033 --> 01:10:49,736
PATIENT WHETHER THERE'S A LOT OF

1677
01:10:49,736 --> 01:10:53,039
GENETIC VARIATION AT THAT POINT?

1678
01:10:53,039 --> 01:10:56,643
>> SO THAT -- THE REASON
THAT 

1679
01:10:56,643 --> 01:11:00,413
THAT'S STRIKING AS A SEQUENCE IS

1680
01:11:00,413 --> 01:11:02,048
BECAUSE THERE'S DEFINITELY -- 

1681
01:11:02,048 --> 01:11:03,516
AND THE STRUCTURE WOULD DO A 

1682
01:11:03,516 --> 01:11:07,854
GOOD JOB OF EXPLAINING WHY, BUT 

1683
01:11:07,854 --> 01:11:09,122
THERE'S DEFINITELY SEVERAL 

1684
01:11:09,122 --> 01:11:11,324
RESIDUES WITHIN THE ALIX-BINDING

1685
01:11:11,324 --> 01:11:15,562
MOTIF THAT ARE NOT CONSERVED, 

1686
01:11:15,562 --> 01:11:18,298
BUT THE S40, THE STEERING AT THE

1687
01:11:18,298 --> 01:11:21,101
40 POSITION IS HIGHLY CONSERVED.

1688
01:11:21,101 --> 01:11:23,703
IT'S MOST CONSERVED OTHER THAN 

1689
01:11:23,703 --> 01:11:25,005
THE Y IN THERE, YEAH.

1690
01:11:25,005 --> 01:11:29,042
IF YOU MUTATE IT BY ITSELF, YOU 

1691
01:11:29,042 --> 01:11:32,212
WON'T LOSE ALIX BINDING ABILITY.

1692
01:11:32,212 --> 01:11:34,314
>> UNLESS WE HAVE A LOT OF
GAG 

1693
01:11:34,314 --> 01:11:36,816
SEQUENCES GENERATING A GOOD BIT 

1694
01:11:36,816 --> 01:11:37,584
OF [ INDISCERNIBLE ]. 

1695
01:11:37,584 --> 01:11:38,551
>> YES, YES.

1696
01:11:38,551 --> 01:11:39,719
I THINK IT'S FASCINATING, 

1697
01:11:39,719 --> 01:11:43,123
ACTUALLY.

1698
01:11:43,123 --> 01:11:47,794
>> IT'S ONE OF THE TOOLS
THAT --

1699
01:11:47,794 --> 01:11:51,898
SO ALIX SHOWS IT WILL REDUCE 

1700
01:11:51,898 --> 01:11:53,199
VIRUS REPRODUCTION AND PRAZOLES 

1701
01:11:53,199 --> 01:11:54,467
WILL DO THAT.

1702
01:11:54,467 --> 01:11:55,902
DO YOU KNOW IF THERE'S ANY 

1703
01:11:55,902 --> 01:11:57,637
SYNERGY BETWEEN THOSE TWO OR 

1704
01:11:57,637 --> 01:11:58,438
ADDITIVE EFFECTS?

1705
01:11:58,438 --> 01:12:01,641
>> THE QUESTION IS, IF I
HEARD 

1706
01:12:01,641 --> 01:12:05,679
IT RIGHT, DID YOU SAY STATS, 

1707
01:12:05,679 --> 01:12:09,649
STATINS AND PRAZOLES WILL 

1708
01:12:09,649 --> 01:12:11,785
BOTH -- I MISSED THE VERB. 

1709
01:12:11,785 --> 01:12:14,754
>> IS IT AN INDEPENDENT 

1710
01:12:14,754 --> 01:12:15,722
MECHANISM, BOTH THROUGH TSG101, 

1711
01:12:15,722 --> 01:12:17,657
WILL THEY -- IS THERE A SYNERGY 

1712
01:12:17,657 --> 01:12:18,358
BETWEEN THOSE TWO?

1713
01:12:18,358 --> 01:12:20,193
>> SO WE DON'T KNOW.

1714
01:12:20,193 --> 01:12:20,860
I DON'T KNOW, YEAH.

1715
01:12:20,860 --> 01:12:25,832
WE SHOULD ASK THAT.

1716
01:12:25,832 --> 01:12:30,003
>> WE HAVE PATIENTS ON
SOME, BUT

1717
01:12:30,003 --> 01:12:32,972
BOTH, SO THIS EARLY LATE DOMAIN 

1718
01:12:32,972 --> 01:12:35,175
DEFECT THAT YOU SEE WITH THE 

1719
01:12:35,175 --> 01:12:36,776
PRAZOLES, MECHANISTICALLY, WHAT 

1720
01:12:36,776 --> 01:12:37,877
DO YOU THINK IS HAPPENING THERE?

1721
01:12:37,877 --> 01:12:41,648
IT SEEMS NOT TO BE A CLASSIC 

1722
01:12:41,648 --> 01:12:44,150
TSG101 IMPAIRMENT SORT OF LATE 

1723
01:12:44,150 --> 01:12:47,454
DOMAIN, BUT MORE OF AN ASSEMBLY 

1724
01:12:47,454 --> 01:12:49,889
LATTICE CONSTRUCTION EFFECT.

1725
01:12:49,889 --> 01:12:51,424
WHAT DO YOU THINK 

1726
01:12:51,424 --> 01:12:52,392
MECHANISTICALLY IT'S HAD?

1727
01:12:52,392 --> 01:12:54,194
>> THAT IS ACTUALLY A
QUESTION 

1728
01:12:54,194 --> 01:12:55,695
THAT IS, TO ME -- THE QUESTION 

1729
01:12:55,695 --> 01:12:57,997
WAS, WHAT DO I THINK IS ACTUALLY

1730
01:12:57,997 --> 01:12:59,566
HAPPENING BECAUSE THE PRAZOLES 

1731
01:12:59,566 --> 01:13:03,369
SORT OF ARREST AT AN EARLY 

1732
01:13:03,369 --> 01:13:04,504
STAGE, EARLIER THAN WHEN YOU 

1733
01:13:04,504 --> 01:13:07,941
WOULD CALL ESCRT TO COME FINISH 

1734
01:13:07,941 --> 01:13:11,344
THE JOB OFF AND SNIP THE NET.

1735
01:13:11,344 --> 01:13:15,181
AND SO IT HAS -- WHAT IT SAYS TO

1736
01:13:15,181 --> 01:13:18,752
US IS THAT VERY EARLY ON, WHAT 

1737
01:13:18,752 --> 01:13:24,824
YOU NEED IS A UBIQUITIN NASCENT 

1738
01:13:24,824 --> 01:13:26,392
EVENT, THAT UBIQUITIN EVENT -- 

1739
01:13:26,392 --> 01:13:29,162
ON TUESDAYS AND THURSDAYS, WE 

1740
01:13:29,162 --> 01:13:30,230
THINK THE UBIQUITIN NATION EVENT

1741
01:13:30,230 --> 01:13:31,297
IS FOR SCAFFOLD.

1742
01:13:31,297 --> 01:13:32,832
ON MONDAYS AND WEDNESDAYS, WE 

1743
01:13:32,832 --> 01:13:37,704
THINK, NO, NO, NO, IT HAS TO DO 

1744
01:13:37,704 --> 01:13:39,672
WITH WHICH ESCRT MIGHT IN FACT 

1745
01:13:39,672 --> 01:13:42,342
BE, YOU KNOW, SORT OF LIKE -- 

1746
01:13:42,342 --> 01:13:45,011
YOU GOT A BRILLIANT EARLY TO 

1747
01:13:45,011 --> 01:13:48,748
ACTIVATE IT, BUT THE OTHER 

1748
01:13:48,748 --> 01:13:50,884
ASPECT IS THAT THE -- EVEN WITH 

1749
01:13:50,884 --> 01:13:53,052
RESPECT TO ALIX THAT WE WERE 

1750
01:13:53,052 --> 01:13:55,355
JUST TALKING ABOUT, IT HAS GOT 

1751
01:13:55,355 --> 01:13:57,190
TO BE UBIQUITINATED TO FUNCTION 

1752
01:13:57,190 --> 01:13:59,192
AS WELL.

1753
01:13:59,192 --> 01:14:01,161
AND SO IT COULD BE A THIRD 

1754
01:14:01,161 --> 01:14:03,229
MECHANISM WOULD BE THAT EVEN IN 

1755
01:14:03,229 --> 01:14:06,432
YOUR ADAPTOR MACHINERY, THE 

1756
01:14:06,432 --> 01:14:08,935
UBIQUITINATION NEEDS TO BE THERE

1757
01:14:08,935 --> 01:14:10,170
EITHER BECAUSE THAT WHOLE -- 

1758
01:14:10,170 --> 01:14:11,938
SOME INTERACTION IN PLACE FOR 

1759
01:14:11,938 --> 01:14:13,673
SOME TIME OR NOT.

1760
01:14:13,673 --> 01:14:15,341
SO DEPENDING ON THE DAY OF THE 

1761
01:14:15,341 --> 01:14:16,810
WEEK, WE'VE DIFFERENT IDEAS 

1762
01:14:16,810 --> 01:14:17,877
ABOUT, YOU KNOW, WHY IT MIGHT BE

1763
01:14:17,877 --> 01:14:22,515
GOING ON.

1764
01:14:22,515 --> 01:14:26,052
>> WE HAVE AN ONLINE
QUESTION.

1765
01:14:26,052 --> 01:14:27,887
WONDERFUL TALK, THANK YOU.

1766
01:14:27,887 --> 01:14:30,390
WHY DO YOU THINK THERE'S SUCH A 

1767
01:14:30,390 --> 01:14:36,596
DRAMATIC DIFFERENCE IN HOW THE 

1768
01:14:36,596 --> 01:14:37,797
140A/F MUTATIONS AFFECT VIRAL 

1769
01:14:37,797 --> 01:14:39,399
FITN DEPENDING SPECIFICALLY ON 

1770
01:14:39,399 --> 01:14:40,667
WHETHER THERE IS OR NOT DRUG 

1771
01:14:40,667 --> 01:14:41,334
PRESSURE APPLIED?

1772
01:14:41,334 --> 01:14:43,536
>> YES, SO WHAT I'M
WONDERING 

1773
01:14:43,536 --> 01:14:45,672
ABOUT IS THIS AND IN A WAY, IT'S

1774
01:14:45,672 --> 01:14:46,840
THE DRIVING FORCE FOR THE 

1775
01:14:46,840 --> 01:14:49,042
EXPERIMENTS THAT WE'RE TRYING TO

1776
01:14:49,042 --> 01:14:49,742
FINISH UP RIGHT NOW.

1777
01:14:49,742 --> 01:14:51,845
SO ONE OF THE THINS THAT WE KNOW

1778
01:14:51,845 --> 01:15:01,688
IS, IF YOU PUT THE MUTATION IN T

1779
01:15:01,688 --> 01:15:02,989
CELLS, THEN IT'S VERY POOR, AS 

1780
01:15:02,989 --> 01:15:04,123
THE PERSON JUST SAID.

1781
01:15:04,123 --> 01:15:10,363
IF IN FACT YOU PUT IT IN JERCA, 

1782
01:15:10,363 --> 01:15:11,331
LOVES IT.

1783
01:15:11,331 --> 01:15:13,266
THAT'S THE REASON WHY WE'RE 

1784
01:15:13,266 --> 01:15:14,601
EXPLORING ALL THESE DIFFERENT 

1785
01:15:14,601 --> 01:15:15,635
CELL ENVIRONMENTS.

1786
01:15:15,635 --> 01:15:18,071
SO I'M WONDERING WHETHER, AND 

1787
01:15:18,071 --> 01:15:20,540
THIS MAY GET US NORTHERN, BUT 

1788
01:15:20,540 --> 01:15:21,841
WHAT WE'RE -- GET US NOWHERE, 

1789
01:15:21,841 --> 01:15:25,144
BUT WHAT WE'RE TRYING TO DO IS 

1790
01:15:25,144 --> 01:15:26,479
IDENTIFY WHAT MAYBE THE PLAYERS 

1791
01:15:26,479 --> 01:15:30,116
IN THE PHENOTYPE THAT IT 

1792
01:15:30,116 --> 01:15:33,019
MANIFESTS AND TO ASK WHETHER OR 

1793
01:15:33,019 --> 01:15:34,320
NOT THERE'S DIFFERENTIAL 

1794
01:15:34,320 --> 01:15:36,222
EXPRESSION OF FORMS OF THOSE IN 

1795
01:15:36,222 --> 01:15:38,892
THESE VARIOUS CELL ENVIRONMENTS.

1796
01:15:38,892 --> 01:15:40,593
IT COULD BE AS, QUOTE-UNQUOTE, 

1797
01:15:40,593 --> 01:15:41,694
SIMPLE AS THAT.

1798
01:15:41,694 --> 01:15:44,130
AT LEAST THAT'S WHERE WE 

1799
01:15:44,130 --> 01:15:44,464
STARTING.

1800
01:15:44,464 --> 01:15:48,835
>> DO YOU FIND THAT
CLEAVAGE 

1801
01:15:48,835 --> 01:15:51,704
WITH A PROTEASE INHIBITOR 

1802
01:15:51,704 --> 01:15:54,941
RESISTANT PROTEASE AT THAT SITE,

1803
01:15:54,941 --> 01:15:56,509
THAT IT'S AFFECTED BY THAT 

1804
01:15:56,509 --> 01:15:58,645
MUTATION?

1805
01:15:58,645 --> 01:16:01,681
WE'RE SEEING THAT WITH A 

1806
01:16:01,681 --> 01:16:02,081
[ INDISCERNIBLE ].

1807
01:16:02,081 --> 01:16:03,383
>> YEAH.

1808
01:16:03,383 --> 01:16:07,720
SO NOT AS FAR AS WE CAN TELL.

1809
01:16:07,720 --> 01:16:11,658
SO IT WAS ON THE SLIDE, I THINK,

1810
01:16:11,658 --> 01:16:17,163
BUT THERE WAS A -- THERE WAS -- 

1811
01:16:17,163 --> 01:16:22,268
WHAT WE DID WAS WE TOOK FROM THE

1812
01:16:22,268 --> 01:16:24,604
ONE DATABASE MUTATIONS THAT CAME

1813
01:16:24,604 --> 01:16:27,941
UP IN OTHER INDIVIDUALS AS 

1814
01:16:27,941 --> 01:16:29,008
CONFERRING SOME DEGREE OF 

1815
01:16:29,008 --> 01:16:30,643
PROTEASE RESISTANCE.

1816
01:16:30,643 --> 01:16:32,779
AND WE ASKED WHETHER IN FACT 

1817
01:16:32,779 --> 01:16:33,680
THAT WAS INFLUENTIAL.

1818
01:16:33,680 --> 01:16:35,181
AND WE DIDN'T SEE ANYTHING THAT 

1819
01:16:35,181 --> 01:16:38,184
WE COULD DETECT.

1820
01:16:38,184 --> 01:16:40,954
>> ANYBODY ELSE?

1821
01:16:40,954 --> 01:16:44,590
>> I JUST WONDER IS THERE
A WAY 

1822
01:16:44,590 --> 01:16:48,361
TO TEST ON THE SAMPLES 

1823
01:16:48,361 --> 01:16:49,629
[ INDISCERNIBLE ] THE EXTRA 

1824
01:16:49,629 --> 01:16:53,066
RECEPTOR ON THE CELLS THAT WE 

1825
01:16:53,066 --> 01:16:54,934
CANNOT NOW COUNT.

1826
01:16:54,934 --> 01:16:58,738
>> THAT'S AN INTERESTING
ONE.

1827
01:16:58,738 --> 01:17:00,340
WOULD THAT BE SUFFICIENT?

1828
01:17:00,340 --> 01:17:02,241
>> I SAW SOME STUDY
LOOKING FOR 

1829
01:17:02,241 --> 01:17:04,677
THE ROLE OF ESTROGEN ON HIV AND 

1830
01:17:04,677 --> 01:17:06,412
THAT'S WHAT THEY DID.

1831
01:17:06,412 --> 01:17:08,915
I'M NOT SURE ABOUT THE 

1832
01:17:08,915 --> 01:17:12,151
[ INDISCERNIBLE ] SOME POTENTIAL

1833
01:17:12,151 --> 01:17:13,386
SEX-RELATED.

1834
01:17:13,386 --> 01:17:15,421
>> SO THAT'S SORT OF LIKE 

1835
01:17:15,421 --> 01:17:16,823
TEASING -- OF COURSE, IF YOU'RE 

1836
01:17:16,823 --> 01:17:19,726
GOING TO KNOCK DOWN, THEN THE 

1837
01:17:19,726 --> 01:17:21,060
RECEPTOR, IF YOU JUST PLUG IT 

1838
01:17:21,060 --> 01:17:23,997
INTO THE RECEPTOR, PERIOD, WELL,

1839
01:17:23,997 --> 01:17:27,433
ONE OF THE JOBS OF ESCRT 

1840
01:17:27,433 --> 01:17:30,003
MACHINERY IS TO DOWN-REGULATE 

1841
01:17:30,003 --> 01:17:30,336
RECEPTORS.

1842
01:17:30,336 --> 01:17:31,337
I DON'T KNOW IF THAT'S ONE OF 

1843
01:17:31,337 --> 01:17:34,574
THEM, BUT IT WOULD JUST TIE IN 

1844
01:17:34,574 --> 01:17:36,976
WHAT ESCRTs, SO IF IN FACT THERE

1845
01:17:36,976 --> 01:17:38,711
WASN'T A GENDER PLAY, I DON'T 

1846
01:17:38,711 --> 01:17:40,947
KNOW, BUT THAT'S WHAT SORT OF 

1847
01:17:40,947 --> 01:17:42,749
LIKE STRIKES ME COULD BE AN 

1848
01:17:42,749 --> 01:17:43,416
INTERESTING CONNECTION WITH THAT

1849
01:17:43,416 --> 01:17:49,122
IDEA ANYWAY.

1850
01:17:49,122 --> 01:17:52,025
>> DO YOU HAVE A SENSE OF
TSG101

1851
01:17:52,025 --> 01:17:53,559
IS DIFFERENTIALLY EXPRESSED 

1852
01:17:53,559 --> 01:17:56,729
BETWEEN DIFFERENT AGE GROUPS?

1853
01:17:56,729 --> 01:17:58,131
>> DIFFERENT AGE GROUPS,
YOU 

1854
01:17:58,131 --> 01:17:59,065
MEAN PEOPLE AGE GROUPS?

1855
01:17:59,065 --> 01:18:01,667
HMM.

1856
01:18:01,667 --> 01:18:04,537
I DON'T KNOW THAT WE WOULD HAVE 

1857
01:18:04,537 --> 01:18:08,341
A WAY OF UNDERSTANDING THAT.

1858
01:18:08,341 --> 01:18:12,612
THE QUESTION WAS, WOULD TSG101 

1859
01:18:12,612 --> 01:18:15,782
BE -- RESTATE THE QUESTION.

1860
01:18:15,782 --> 01:18:18,484
>> DO YOU KNOW IF THERE IS

1861
01:18:18,484 --> 01:18:20,520
DIFFERENTIAL EXPRESSION OF 

1862
01:18:20,520 --> 01:18:22,488
TSG101 AMONG DIFFERENT HUMAN AGE

1863
01:18:22,488 --> 01:18:22,722
GROUPS?

1864
01:18:22,722 --> 01:18:25,291
>> I WOULD ASSUME THAT THE

1865
01:18:25,291 --> 01:18:27,527
ANSWER IS NO.

1866
01:18:27,527 --> 01:18:28,961
IT'S UBIQUITOUSLY EXPRESSES.

1867
01:18:28,961 --> 01:18:31,431
I MEAN, PLANTS EVEN HAVE IT.

1868
01:18:31,431 --> 01:18:33,733
IT'S EMBRYOTICALLY LETHAL, SO 

1869
01:18:33,733 --> 01:18:35,401
EVERYBODY NEEDS IT, BUT I HAVE 

1870
01:18:35,401 --> 01:18:37,970
NO INDICATION -- SO IN THE 

1871
01:18:37,970 --> 01:18:39,906
THESIS OF GRADUATE STUDENTS WHO 

1872
01:18:39,906 --> 01:18:43,843
WORKED ON THIS INITIALLY IS A 

1873
01:18:43,843 --> 01:18:46,379
BUNCH OF DIFFERENT TISSUES, 

1874
01:18:46,379 --> 01:18:49,215
CERTAINLY, BUT IN TERMS OF AGE, 

1875
01:18:49,215 --> 01:18:50,383
WE NEVER LOOKED.

1876
01:18:50,383 --> 01:18:51,451
I DON'T SEE A REASON WHY IT 

1877
01:18:51,451 --> 01:18:53,152
SHOULD.

1878
01:18:53,152 --> 01:18:55,054
BUT WHO KNOWS?

1879
01:18:55,054 --> 01:18:57,423
>> AND ONE MORE QUESTION
IF 

1880
01:18:57,423 --> 01:18:57,757
THERE'S TIME.

1881
01:18:57,757 --> 01:18:59,992
DO WE HAVE TIME?

1882
01:18:59,992 --> 01:19:01,394
YES?

1883
01:19:01,394 --> 01:19:02,395
OKAY.

1884
01:19:02,395 --> 01:19:02,895
>> SURE. 

1885
01:19:02,895 --> 01:19:05,998
>> DO YOU THINK, ARE YOU
OR 

1886
01:19:05,998 --> 01:19:07,900
ANYONE ELSE LOOKING AT THE 

1887
01:19:07,900 --> 01:19:10,736
EFFECT THAT PRAZOLES MIGHT HAVE 

1888
01:19:10,736 --> 01:19:12,538
ON OTHER UBIQUITIN LIGASES, LIKE

1889
01:19:12,538 --> 01:19:14,707
THE ONES THAT ACTUALLY HAVE THE 

1890
01:19:14,707 --> 01:19:16,676
ABILITY TO TRANSFER?

1891
01:19:16,676 --> 01:19:17,510
>> YES.

1892
01:19:17,510 --> 01:19:18,778
SO THAT'S SOMETHING WE'RE DOING 

1893
01:19:18,778 --> 01:19:19,445
RIGHT NOW.

1894
01:19:19,445 --> 01:19:22,381
IT TURNS OUT THAT TSG IS ABLE TO

1895
01:19:22,381 --> 01:19:25,118
PROTECT A WHOLE BUNCH OF 

1896
01:19:25,118 --> 01:19:26,252
DIFFERENT ONES.

1897
01:19:26,252 --> 01:19:29,188
NOT EVERYBODY, ALL RIGHT, BUT A 

1898
01:19:29,188 --> 01:19:32,725
WHOLE BUNCH OF DIFFERENT ONES, 

1899
01:19:32,725 --> 01:19:35,461
AND THE PRAZOLES WOULD APPEAR TO

1900
01:19:35,461 --> 01:19:36,129
DO SO.

1901
01:19:36,129 --> 01:19:37,864
I'LL PUT AN ASTERISK THERE FOR 

1902
01:19:37,864 --> 01:19:39,332
TWO REASONS.

1903
01:19:39,332 --> 01:19:40,299
NUMBER ONE, BECAUSE WE WERE 

1904
01:19:40,299 --> 01:19:42,268
DOING THE EXPERIMENT IN THE HOPE

1905
01:19:42,268 --> 01:19:46,239
OF GETTING A HANDLE ON HOW IT 

1906
01:19:46,239 --> 01:19:49,175
ACTUALLY IS MANIPULATING THE 

1907
01:19:49,175 --> 01:19:50,443
UBIQUITINATION EVENT.

1908
01:19:50,443 --> 01:19:51,577
BECAUSE -- AND SO THE REASON I 

1909
01:19:51,577 --> 01:19:54,580
PUT AN ASTERISK IS WE'VE ONLY 

1910
01:19:54,580 --> 01:19:55,715
DONE THE EXPERIMENT WITH A 

1911
01:19:55,715 --> 01:19:59,318
LIMITED NUMBER OF THEM AND SO WE

1912
01:19:59,318 --> 01:20:01,687
HAVEN'T TAKEN ONES LIKE -- YOU 

1913
01:20:01,687 --> 01:20:03,422
KNOW, ACROSS THE BOARD, 

1914
01:20:03,422 --> 01:20:04,257
DIFFERENT BY OTHER INDICATIONS 

1915
01:20:04,257 --> 01:20:07,160
AND TRIED IT.

1916
01:20:07,160 --> 01:20:08,828
>> ALL RIGHT.

1917
01:20:08,828 --> 01:20:18,828
WELL, THANK YOU VERY MUCH.