GOOD AFTERNOON.
WELCOME TO THE WEDNESDAY
AFTERNOON LECTURE SERIES, MY
NAME IS ALEXANDER ZALAZAR, AND I
AM CHIEF OF THE NETWORK FUNCTION
OF NISB OF NIAID AND TODAY I
HAVE THE HONOR TO INTRODUCE
TODAY'S WALS SPEAKER
DR. ILEANA CRISTEA, BUT FIRST I
HAVE SOME INFORMATION, THE CME
CODE FOR TODAY IS 37953.
I REPEAT, 37953.
ALSO IF YOU CAN TELL THIS IS A
HYBRID LECTURE IN WHICH SOME OF
US ARE HERE AND THE VAST
MAJORITY OF YOU ARE WATCHING NIH
VIDEOCAST.
THOSE THAT ARE REMOTE CAN STILL
ASK QUESTIONS, CAN YOU DO SO BY
CLICKING ON THE BUTTON JUST
BELOW YOUR VIDEOCAST WINDOW THAT
SAYS SEND LIVE FEEDBACK, CLICK
ON THAT IN YOUR NAME, END
QUESTION AND WE WILL RELAY TO
DR. CRISTEA AT THE END OF HER
LECTURE.
CAN YOU SUBMIT QUESTIONS
ANYTIME.
PLEASE NOTE THAT TODAY'S LECTURE
IS THE FINAL OF THIS WALS
SEASON, WE'LL WILL TAKE A 2
MONTH BREAK AND BEGIN A NEW
SIEWZON ON SEPTEMBER 7th WHERE
WE ARE JOINED BY DR. JILL
[INDISCERNIBLE]OT TOPIC OF
MICROBIAL DISEASE.
OUR SPEAKER TODAY IS
DR. ILEANA CRISTEA, SHE IS WITH
THE LABORATORY CELLULAR
MECHANISMS AT PRINCE ON TON
UNIVERSITY, AS WELL AS
MECHANISMS USED BY VIRUSES TO
INHIBIT OR HIJOCK HOST CELL
PROCESSES.
HER LAB EMPLOYEES
MULTIDISCIPLINARY APPROACHES
INTEGRATING MOLECULAR VIROLOGY,
MICROSCOPY, MASS
SPECTROMETRY-BASED PROTEOMICS
AND BIO-INFORMATICS.
IN RECEIPT YEARS, SHE DEVELOPED
SPACIAL AND TEMPORAL VITALS FOR
HOST PROTEIN INTERACTIONS,
BRIDGING DEVELOPMENT TO
IMPORTANT FINDINGS IN BIOLOGY,
FOR EXAMPLE HER LABORATORY HAS
CONTRIBUTED TO THE EMERGENCE OF
THE RESEARCH FIELD OF NUCLEIC
DNA SENSING AND UNCOVERING
MECHANISMS UNDERLYING MODELING
AND ORGANELLE STUDY STRUCTURE
FUNCTION RELATIONSHIPS DURING
INFECTIONS, AND THEY HAVE
DISCOVERED BROAD SPECTRUM
ANTIVIRAL FACTORS.
ILEANA RECEIVED HER MEDICAL
DEGREE IN MACROCHEMMISTRY
FOLLOWED BY A Ph.D. FROM
MANCHESTER UNIVERSITY.
AND SHE THEN PURSUED POST
DOCTORAL WORK AT THE LABORATORY
AT ROCKEFELLER UNIVERSITY.
ILEANA JOINED PRINCETON
UNIVERSITY IN 2008 AS ASSISTANT
PROFESSOR AND BECAME ASSOCIATE
PROFESSOR IN 2013 AND ATTAINED
HER CURRENT POSITION IN TWEBT
16.
ELEANA IS VERY ACTIVE IN THE
PROTEOMICS COMMUNITY.
SHE'S THE PAST PRESIDENT OF THE
HUMAN PROTEOME ORGANIZATION OF
[INDISCERNIBLE], PAST CHEAR OF
THE HUMAN PROTEOME PROJECT AND
CHAIR OF THE INFECTIOUS DISEASE
TEAM OF [INDISCERNIBLE] Ph.D.
SHE HAS WORKED AT THE COLD
SPRINGS LABORATORY FOR MORE THAN
10 YEARS AND SENIOR EDITOR FOR
END SYSTEMS, ASSOCIATE EDITOR
FOR THE PROTEOME RESEARCH AND
UNDER THE MOLECULAR SYSTEMS
BIOLOGY AND MOLECULAR AND
CELLULAR PROTEOMICS.
EASE ALSO REYIPIENT OF MANY
HONORS AND,A WARDS WHICH INCLUDE
THE [INDISCERNIBLE] PRIZE FROM
THE BRITISH MASS SPECTROMETRY
SOCIETY, NIH NIDA AVAN GUARD
ROAD FOR RESEARCH, HUMAN FRAN
TEARS HUMAN INVESTIGATOR AWOORD,
CAREER AWARD FROM ICS,
[INDISCERNIBLE] SKOALAR AWARD
AND THE MOST PROUDLY THE
PRINCETON UNIVERSITY GRADUATE
MOLECULAR AWARD.
THE TITLE OF HER TALK TODAY IS
VIRAL ORGANELLE REMODELING FOR
HOST CELL BIOLOGY.
ILEA, INA, WELCOME AND THANK YOU
FOR BEING WITH US TODAY [
APPLAUSE ]
>> THANK YOU SO MUCH, THANK YOU
FOR THE VERY KIND INTRODUCTION
AND THANK YOU FOR THE
INVITATION, IT'S A GREAT
PLEASURE TO BE HERE AND I THANK
YOU ALL AND THE ORGANIZERS AND
THANK YOU FOR BEING IN PERSON
AND ALSO I THANK THOSE ONLINE
FOR JOINING IN TODAY.
I HAVE TO SAY IT'S SO GREAT TO
BE IN PERSON FOR A SEMINAR IN 3D
SO IT'S WAY HUGE DIFFERENCE AND
GREAT HONOR TO BE HERE.
SO AS YOU HEARD MY LAB IS AT
PRINCETON IN THE DEPARTMENT OF
BIOLOGY WHERE WE INDGREAT
BIOLOGY WITH PROTEOMICS TO STUDY
AT THE LEVEL OF HUMAN VIRUSES
AND TODAY I SELECTED TO TELL YOU
ABOUT SOME OF OUR SCIENTIFIC
ADVENTURES DURING THE PANDEMIC
THAT HAVE BROUGHT US TO BECOME
QUITE INTERESTED AND REALLY
FASCINATED WITH THIS CONCEPT OF
DYNAMIC ORGANELLE REMODEL
THAGOREAN HAPPENS DURING THE
REPLICATION CYCLE OF VIRAL
INFECTIONS SO I'LL TELL YOU MORE
ABOUT THAT AND PUT THIS IN
BROADER CONTEXT, BUT FIRST I
WILL TELL YOU ABOUT MY
DISCLOSURES, I AM THE FOUNDER
AND CONFOUNDER OF A COMPANY THAT
MAKES ANTIVIRAL TREATMENTS BY
TARGETING SERIES POINTS TUEINS
AND I'M ALSO A MEMBER OF
SCIENTIFIC ADVISORY BOARD OF 908
DEVICES WHICH IS A MASS
SPECTROMETRY DEVICES, WHAT I
WILL SPEAK ABOUT TODAY IS NOT
CONTECTED TO THESE 2 COMPANIES
BUT JUST TO LET YOU KNOW.
ALL RIGHT, SO TO START WITH
BROADER TOPIC, THE CELL IS
DEPENDENTOT ABILITY OF THE VIRUS
TO REMODEL ORGANELLES, SO IN
OTHER WORDS THE CHANGES THAT CAN
OCCUR IN ORGANELLE SHAPE AND
COMPOSITION AND FUNCTION ARE
ABSOLUTELY ESSENTIAL FOR THE
VIRAL INFECTIONS, FOR THE VIRAL
REPLICATION CYCLES AND 1
IMPORTANT ILLUSTRATION OF THIS
IS GIVEN BY INFECTION WITH HUMAN
CYTOMEGALOW VIRUS, THIS IS A
BETTER PERP ESTIMATE THAD VIRUS
THAT IS WIDELY SPREAD, IT HAS A
LARGE GENOME AND QUITE A LENGTHY
REPLICATION CYCLE OF 4-5 DAYS,
SO THIS HAS BOTH THE GENETIC
CAPACITY AS WELL AS TIME TO
INDUCE STRIKING CHANGES IN THE
SUBCELLULAR ENVIRONMENT DURING
ITSELF REPLICATION CYCLE SO FOR
EXAMPLE FOR ENTRY, IT REMODELS
THE PLASMA MEMBRANE, IT REMODELS
CYTOSKELETON AND ENDOSTUDIES OF
MULTIPLE ENDOCRINES AND THEN
FOLLOWING IT'S NUCLEAR
REPLICATION, THE VIRUS DISRUPTS
THE PERIPHERY TO ALLOW THE VIRAL
CAPSID TO EGRESS FROM THE
NUCLEUS.
LATER IN THE INFECTION, THE
VIRUS MAKES A NEW ASSEMBLY, A
SUBSET COMPARTMENT FORMED WITH
GOLGI 'EM BRAINS AND SUBCELLS
AND OVER HERE I WILL SHOW YOU
THIS THAT I MENTIONED EARLIER.
SO THE ASSEMBLY COMPARTMENT THAT
MAKES NEW VIRAL PARTICLES THAT
CONTAIN ENDOSTUDIES OF MULTIPLE
ENDOCRINES IN THEM AND GOLGI
MEMBRANES THAT ARE RESTRUCTURED
AND LATER FOR EGRESS, ALSO IT
HIJACKS TRAFFICKING PROCESSES,
HOST TRAFFICKING PROCESSES,
ANOTHER HALLMARK OF HSV
INFECTION AND MITOCHONDRIAL
INFECTION AND I WILL TELL YOU
ABOUT THIS LATER IN THIS TALK,
HOWEVER WHAT'S NOT FULLY
UNDERSTOOD IS HOW THESE
REMODELING EVENTS ARE CONNECTED
TO SOME OF THE VIRAL
PATHOLOGIES, SO INDUCED BY THIS
VIRUS SO FOR EXAMPLE, CARDIAC
DISEASE OR THE ONCOMODLATTORY
PROPERTIES OF HCMV, HOWEVER,
IT'S IMPORTANT TO NOTE
NAREMODELLING IS NEEDED FOR ALL
VIRUS, SO REGARD WILL OF GENOME
SIZE, REPLICATION STRATEGIES,
DIFFERENT CELL TROAPISMS, ALL
VIRUSES ARE UNITED IN THEIR NEED
FOR REGULATION OF AGER ANLE
FUNCTION, ORGANELLE COMPOSITION
AND SHAPE DURING THE REPLICATION
CYCLE.
SO FOR EXAMPLE, HSV 1, ALSO
HERPES VIRUS ALSO REGULATES THE
EGRESS OF THE VIRAL SID.
AND ALSO [INDISCERNIBLE] THE
CORONA VIRUS, LIKE OTHER BETA
CORONA VIRUSES INDUCE REMODELING
OF THE ER AND MAKES THE DOUBLE
MEMBRANE FOR THE REPLICATION OF
THESE VIRUS, SO NOW ALL THESE
DYNAMIC MODELING EVENTS THAT
OCCUR DURING INFECTION AND THIS
IS SOMETHING WE HAVE FOCUSED ON
IN RECENT YEARS.
SO WE--WITH A QUESTION OF HOW
AND WHY WHICH OF THESE PROCESSES
ARE FOR THE BENEFIT OF VIRUS OR
THE HOST AND IN ORDER TO
UNDERSTAND THIS INFECTION
INDUCED REWIRING OF HOST CELLS,
WE HAVE MULTITUDE OF METHODS
AVAILABLE FROM MOTE I DON'T
MEANICS, FROM LIPID OMICS, AND
TODAY I WILL TRY TO HIGHLIGHT
SOME OF THE STRATEGIES WE HAVE
USED OVER THE YEARS AND IN
PARTICULAR IN MORE RECENT YEARS
AND HOW WE HAVE LEARNED ABOUT
THIS EPITHELIAL RACELLULAR
REORGANIZATION FROM THESE
DIFFERENT METHODS.
SO WE HAVE STARTED INSHALLLY BY
FOCUSING ON UNDERSTANDING THE
VIRUS HOST PROTEIN INTERACTIONS
THAT ARE DRIVING SOME OF THESE
SIGNALING CASCADES AND SOME OF
THESE REMODELING EVENTS IN THE
CELL.
AND OF COURSE, WE HAVE SO MANY
DIFFERENT APPROACHES WE CAN USE
FOR STUDYING PROTEIN
INTERACADEMIESS DURING AN
INFECTION, WE CAN USE AFFINITY
PURIFICATION, CROSS LINKING AND
MANY PROTEIN MICROARRAYS AND
THESE METHODS EVER ARE
PROSCRIEDING US WITH AN
OPPORTUNITY TO STUDY WITH A
LOCAL PERSPECTIVE FOCUSED ON 1
PROTEIN OF INTEREST AT A TIME OR
FROM A GLOBAL PERSPECTIVE AND
GIVEN OUR INTEREST IN ORGANELLE
REMODELING, MORE AND MORE IN
THESE RECENT YEARS WE BECAME
FASCINATED BY THE IDEA OF TRYING
TO CAPTURE IN 1 GOAL, WHAT
HAPPENS IN THE CELL DURING
INFECTION AND HOW THESE EVENTS
COULD BE COORDINATED SO 1 METHOD
I WANT TO HIGHLIGHT TODAY IS
THIS THERMOCOAGULATION PROFILING
AND I JUST TO TAKE YOU THROUGH
THE APPROACH THIS METHOD, SO THE
SAMPLE OF INTEREST WE HAVE USED
THESE WITH CELLS AS WELL AS
TISSUES, THE SAMPLE OF THIS
INTEREST IS SUBJECTS TO
INCREASING TEMPERATURES AND THEN
QUANTITATIVE MASS SPECTROMETRY
IS USED TO QUANTIFY SOLUBLE
PROTEINS AT EACH OF THESE
TEMPERATURES.
IT IS READ OUT IS ACTUALLY AT
COAGULATION CURVES AS THE
TEMPERATURE INCREASES SO HOW DO
WE LEARN FROM THIS ABOUT PROTEIN
INTERACTIONS?
WELL THE PRINCIPLE OF THIS IS
THAT PROTEINS INTERACT WITH EACH
OTHER ARE STABILIZING EACH OTHER
AND THEY MAY HAVE STABILIZE EACH
OTHER AN THE TEMPERATURE
INCREASES AND THIS WORKS QUITE
BEAUTIFULLY OR YOU CAN CAPTURE
OR SCALE IT IN 1 GOING SAMPLE.
SO [INDISCERNIBLE] A POST DOC IN
MY LAB WAS THE FIRST TO APPLY
THIS METHOD TO VIRAL INFECTION
AND HE APPLIED IT TO HCMV
INFECTION AND THAL CASE, HE TOOK
EXACTLY THIS APPROACH BUT
INVESTIGATED SAMPLES AT
DIFFERENT TIME POINTS OF
INFECTION AND BY LOOKING AT THIS
CO AGGREGATION PROTILE AS THE
INFECTION PROGRESSED AND HOW
THIS COAGULATION PROFILES
CHANGE, WE COULD PREDICT WHICH
INTERACTIONS ARE MAINTAINED
DURING INFECTION, WHICH 1S ARE
FORMED ONLY SPECIFICALLY AT A
CERTAIN TIME POINT OF INFECTION,
WHICH 1S ARE DISRUPTED AS THIS
PROGRESSING AND MORE RECENTLY WE
HAVE FURTHER DEVELOPED THIS
APPROACH AND JOSHUA JUSTICE IN
MY LAB APPLIED IT FOR STUDYING
HSV1 INFECTION.
TRAFFIC REED, THE Ph.D.
STUDENT NOW HAS DEVELOPED
MACHINE LEARNING ALGORITHMS AND
FURTHER ADVANCED THIS APPROACH
IN ORDER TO BE ABLE TO STUDY
DIFFERENT TYPES OF PROTEIN
CLASSES LIKE MEMBRANE BOUND
PROTEINS AND PROTEINS THAT ARE
HARDER TO DETECT NORMALLY, BUT I
WANT TO KIND OF GIVE YOU AN IDEA
OF WHAT WE CAN FIND WITH THIS
METHOD.
SO WE CAN OF COURSE MONITOR
PROTEIN COMPLEXES THAT WE KNOW
THAT THEY EXIST, FUNCTIONAL
PROTEIN COMPLEXES THAT HAVE BEEN
REPORTED SUCH AS THOSE THAT ARE
IN THE DATABASE BUT SAME TIME
YOU CAN UNCOVER DE NOVO
INTERACTIONS THAT WERE NOT
PREVIOUSLY KNOWN, EITHER VIRUS
HOST OR HOST-HOST INTERACTIONS
AND I WANT TO GIVE YOU 1 EXAMPLE
FROM 1 OF OUR LONG STANDING
INTERESTS AND WE HAVE LOOKED IN
THIS LAB FOR OVER 14 YEARS AND
HOW WE HAVE GAINED NEW
INFORMATION BY APPLYING THIS
THERMAL COAGULATION PROFILES TO
THE CONCEPT OF NUCLEIC SENSING.
SO UPON NUCLEIC REPLICATING
VIRUSES SUCH AS HSV OR HSV1, IT
TRAFFICS ALONG MICROTUBELES AND
THE I HAVE RID CAPSID NUCLEAR
PORE AND THE VIRAL DNA IS
INSERTED INSIDE THE NUCLEUS
HERE, THIS A MOMENT IN TIME THAT
IS VERY IMPORTANT AND IS SORT OF
DYNAMIC EVENTSA THE NUCLEIC
PERIPHERY THAT ARE IMPORTANT
HOST DEFENSE AGAINST VIRAL
INFECTION AND IS CESSATION OF
INNATE IMMUNE SIGNALING BY
RECOGNITION OF THIS VIRAL DNA BY
NUCLEAR DNA SENSORS AND SOME
YEARS AGO WE CHARACTERIZED THIS
PROTEIN 16 THAT BECAME THE FIRST
KNOWN SENSOR SO VIRAL--HOST
PROTEIN THAT'S ABLE TO RECOGNIZE
VIRAL DNA IN THE NUCLEI IN
INFECTED CELLS AND RECOGNIZE
VIRAL DNA FROM OUR OWN CELL DNA
IN THE SAME COMPARTMENT AND HERE
YOU CAN SEE THE GREEN THAT GOES
TO THE PERIPHERY UPON THE
BINDING OF THE VIRAL CAPSID OF
THE PERIPHERY.
SO THIS WAS QUITE AN INTERESTING
SORT OF TIME IN THIS FIELD OF
DNA SENSING BUT THERE ARE SO
MANY QUESTIONS REMAINING LIKE WE
DO NOT UNDERSTAND HOW SIGNALING
HAPPENS, THE DOWN STREAM OF THIS
BINDING OF THIS TO VIRAL DNA OR
EVEN WHAT MAKES THIS PROTEIN GO
TO THE NUCLEIC PERIPHERY TO BIND
TO VIRAL DNA.
SO FROM THE PROFILES WE
DISCOVERED THIS SENSOR VERY
EARLY IN THE INFECTION INTERACTS
WITH THE COMPLEX THAT HAS THE
KINASE, AND DNA, PK AND THIS
HAPPENS EARLY IN INFECTION, AND
THIS IS CO AGREGATION AND IT'S
LOST AS THE INFECTION PROGRESSES
AND WE WENT ON AND OUR FINDINGS
IN THE INTEREST OF TIME WE FIND
THAT THESE CO LOCALIZE AT THE
NUCLEAR PERIPHERY AND SITES OF
VIRAL GENOME IN THE SPOGZ
POSITION AND FURTHER MORE WE
SHOW THAT IFI16 IS NEEDED FOR
RECRUIT CRUDING THE DNA RESPONSE
KINASE TO INCOMING VIRAL GENOMES
AT THE NUCLEOTIDES CLER
PERIPHERY AND THIS IS THE
PHOSPHORYLATED FORM OF THE
ACTIVATED FORM YOU SEE HERE.
SO THIS RECRUITS THIS KINASE TO
INCOMING GENOMES WE ASKED
WHETHER THIS CAN BE A FEEDBACK
LOOP, AND IF THIS IS A PEPTIDE,
AND WE STUDY THIS
PHOSPHORYLATION STATUS IN A
MULTITUDE OF CONDITIONS EITHER
WITH OR WOULD IT DNA INHIBITION
WITH AND WITHOUT DNA DAMAGE
INDUCING DNA DAMAGE BY TREATMENT
WITH GLEEL CELLS O MICEIN AND
WITH AND WITHOUT HSV INFECTION
AT THESE EARLY TIME POINTS OF
INFECTION AND WHAT WE FOUND IS
INSIDE THAT AND IT
PHOSPHORYLATES SPECIFICALLY THIS
THREA9, THAT IS NEEDED FOR THIS
AND THE HEME DO MAINS THAT BIND
TO VIRAL DNA SO THIS RECRUITS
DNA TO THE NUCLEOTIDES CLER
PERIPHERY WHEN IT BECAME
PHOSPHORYLATED AND THAT IS
REQUIRED FOR THE ANTIVIRAL
FUNCTIONS BECAUSE WHEN WE MUTATE
THAT SIDE, THIS IS LESS ABLE TO
INDUCE INTERFORON RESPONSE AND
ANTIVIRAL CYTOKINE EXPRESSION
UPON INFECTION AND IN AGREEMENT
WITH THIS, IF WE INHIBIT DNAPK,
WE OBSERVE INCREASED VIRAL
TIGHTERS FOR HSV 1, SO SO WHAT
THIS TECHNOLOGY ALLOWED US IS
THE ELEGANCE DANCE BETWEEN THE
SENSING AND DNA DAMAGE RESPONSE
IN PARTICULAR DNA PK WHEREBY
IFI16 GOES THROUGH THE DNA
RECRUITS IT AND PHOSPHORYLATE
ANDS THAT IMPORTANT FOR INDUCING
INNATE IMMUNE SIGNALING.
SO THIS IS 1 XVENL OF THE
DYNAMIC PERIPHERY THAT ACTUALLY
THE REPLODLING OF THE STRUCTURE
OF THE NUCLEAR PERIPHERY IS
LATER INFECTION WHEN IT HAS TO
EGRESS FROM THE CELLS SO IN
ADDITION TO LEARNING ABOUT
PROTEIN INTERACTION, YOU CAN
ACTUALLY LEARN ABOUT HOW
INFECTION CHANGES THE PROPERTY
OF A GIVEN PROTEIN, EITHER VIRAL
OR HOST, AND WHETHER IT'S
AGGREGATION PROFILE STARTS TO
CHANGE AS THE INFECTION
PROGRESSES, SO HERE IS LAMINE ON
B INUNINFECTED CELLS POST TIME
POINT INFECTION AND YOU SEE HOW
THE THERMAL PROFILES, THERMAL
AGGREGATION PROFILES AS THE
TEMPERATURE INCREASES, THEY
CHANGE WITH INFECTION AND THIS
IS IN CONTRAST IF I SAY I SELECT
AND THEN CHANGE AND SHOW A
MEMBER OF THE CORE COMPLEX AND
THE STATE IS FAIRLY STABLE OF
THERMOAGGREGATION PROFILES FOR
THE INFECTION AND FURTHER MORE
IF WE TRY TO FIND OUT WHAT THIS
CHANGE IN A PRO FILES MEANS IT
ACTUALLY REPRESENTS THE
DISRUPTION I SHOW YOU HERE AND
ALSO ASSOCIATION WITH VIRAL
PROTEINS THAT ARE CRITICAL IN
THIS CAPSID EGRESS AND
STABILITY, SO FOR EXAMPLE, LAMIN
B1 HERE HAS SAME AGGREGATION
WITH THIS PROTILE, THAT PLAYS A
ROLE IN VIRION STABILITY, SO
WHEN WE FURTHER INTEGRATED
ANALYSIS OF POST MODIFICATION ON
TOP OF THIS KNOWLEDGE OF
INTERACTION, WE STARTED TO FIND
OUT OTHER REGULATORY MECHANISMS
THAT ARE CRITICAL FOR OR
CHANGING THE STRUCTURE OF THE
NUCLEAR PERIPHERY AND LAURA WHO
WAS A Ph.D. STUDENT IN MY LAB,
SHE DISCOVERED LYSEEN
ACETYLATION THAT IS SHOWN ON B1,
WHICH SHOWS WHEN B1 BECOMES
ACETALATED, OF K134, THE
INTEGRITY OF THE NUCLEUS IS
RESTORED AND THE VIRUS IS NO
LONGER TO CREATE THESE
INFOLDINGS AND THIS RESULT
INDEED SEQUESTRATION OF THE
VIRAL DNA INSIDE THE NUCLEUS AND
VIRAL CAPSID INSIDE THE NUCLEUS
AND REDUCED VIRUS PARTICLES BOTH
HERE INTRACELLULARLY AND EXTRA
CELLULARLY AS SHOWN HERE.
BUT WHAT THEY WERE ABLE TO SHOW
IS MORE BROADLY RELEVANT SO
FIRST OF ALL SHE SHOWED THAT
DURING INFECTION OF HSV 1,
ANOTHER NUCLEAR REPLICATING
VIRUS AND ALPHA VIRUS.
AGAIN OF THE NUCLEAR PERIPHERY
OF THE LAMIN B1 PROTECTS THE
INTEGRITY OF THE PERIPHERY AND
DOES NOT OOH LOW TO PROGRESS
FROM THE EGRESS.
AND THEN SHE SHOWED THAT THIS IS
BROADLY RELEVANT IN UNINFECTED
CELLS AND A MECHANISM THAT CAN
CONTROL DNA DAMAGE REPAIR CHOICE
AND CELL CYCLE PROGRESSION.
AND WHEN THIS IS ACETALATED,
THIS PROMOTES INTRA LAMINAR
INTERACTIONS AND THAT'S STRONGER
BUT THAT LEADS THE CHROMA TEEN
TO BE EXCLUDED FROM IT'S
INTERACTIONS WITH THE LAMINA,
IRPT ACTIONS THAT ARE REALLY
CRITICAL FOR DNA DAMAGE REPAIR
AND THEN SHE SHOWED THAT THAT
LEADS TO IMPAIRED RECRUITMENT OF
IMPORTANT DNA REPAIR PROTEINS TO
DOUBLE STRANDED BREAK REPAIRS
SUCH AS 53 BP1 IN PARTICULARLY
AND THAT LEADS TO A SLOWER DNA
REPAIR AND ALSO PERSISTENT CELL
CYCLE ACTIVATION OF THE G1 TO S
CONTROL AND PATHWAY CHOICE IN
S-PHASE SO FOR HOMOLOGOUS
RECOMBINATION VERSUS HOMOLOGOUS
ENJOINING, SO IT'S AN EXAMPLE OF
HOW BY STUDYING REMODELING EVENT
DURING INFECTION, WE FOUND A
MECHANISM THAT IS MORE BROADLY
REGULATED AND IMPORTANT DURING
CELL CYCLE PROGRESSION AND
REGULATING THE CHOICE OF DNA
PATHWAY, OF DNA REPAIR PATHWAY.
OKAY, SO, I SHOWED YOU HOW WE
ICE THE ISHT INTERACTION STUDIES
EITHER TARGETED OR GLOBAL TO
LEARN ABOUT DYNAMIC EVENTS THAT
HAPPEN AS THE INFECTION
PROGRESSES.
BUT WE BECAME QUITE MORE AND
MORE INTERESTED IN UNDERSTANDING
REALLY STRUCTURE FUNCTION
RELATIONSHIPS DURING VIRAL
INFECTION, IN ORDER TO
ACCOMPLISH THIS, WE TURNED SOME
YEARS AGO TO SPATIAL PROTEOMICS.
SO I WANT TO SHOW YOU WHERE WE
STARTED AND WHERE WE ARE GOING
NEXT AND I WILL FINISH WITH SOME
WORK THAT IS NOT PUBLISHED AND
THAT WILL TELL YOU ABOUT SOME OF
THE NEW DIRECTIONS IN THE LAB.
SO SOME YEARS AGO WE USED FACIAL
PROT I DON'T MEANICS TO START TO
UNDERSTAND WHAT ARE OTHER
CHANGES THAT HAPPEN IN ORGANELLE
COMPOSITION AND WHAT OTHER VIRAL
PROTEINS THAT MAY BE TARGETING
THESE DIFFERENT ORGANELLES IN
ORDER TO INDUCE THESE CHANGES SO
IN THIS CASE, WE INTEGRATED
MICROSCOPY WHEN WE OBSERVE THE
TIME POINTS WHEN WE OBSERVE THE
STRUCTURAL CHANGES IN AGER ANLES
AND THEN WE COMBINE THEM WITH
ORGANELLE FRACTIONATION AND WE
QUANTITATE THEM WITH MASS TECT
ROMETRY AND THESE DIFFERENT
ORGANELLE FRACTIONS AND USE
THESE SUPER VISED MACHINE
LEARNING TO START TO ASSIGN
DIFFERENT HOSTS AND VIRAL
PROTEINS, 2 DIFFERENT
SUBCELLULAR COMPARTMENTS AS THE
INFECTION PROGRESSES AND THIS IS
REALLY EYE OPENING AT THE TIME,
WE FOUND FRUNKZAL TRANSLOCATIONS
FROM 1 COMPARTMENT TO ANOTHER,
AND WE FOUND CHANGES IN THE AND
1 UNEXPECTED FINDINGS AT THE
TIME THAT ARE PEROX SEEM PROTEIN
INCREASED IN ABUNDANCE DURING
INFECTION AND THIS WAS A COMPANY
TO--WE FOUND OUT BY USING
MICROSCOPY BY A NUMBER OF AND
ALSO THE STRUCTURE THAT WAS
ALTERED DURING INFECTION, SO IN
UNINFECTED CELLS YOU CAN SEE ON
THE LEFT HAND THEY BECOME
FRAGMENTED AND THE 1S THAT
SURVIVE THEY ARE QUITE LARGE AND
WE QUANTIFY THIS TO SHOW THAT
THEY ARE ELONGATED AND CURVED
AND BY INTEGRATED FURTHER,
STRUCTURAL MICROSCOPY AND
MATHEMATICAL MODELS WE LEARN HOW
BIOGENESIS IS REGULATED IN
MAMMALIAN CELLS AND BY USING
PROTEOMICS AND FUNCTION WE
ASSIGN THIS SO SPECIFICALLY WE
FOUND THAT ENVELOPED VIRUSES
SUCH AS HSV 1 INDUCED INCREASE
NUMBER OF PEROXISOMES FOLLOWED
BY CHANGES IN MORPHOLOGY, BUT
THIS IS NOT DRIVEN BY INFECTION
WITH NONENVELOPED VIRUSES, SO
THIS IS REGULATED SO FIRST THE
INFECTION, THE GROWTH AND
INCREASE ABUNDANCE OF PEROXISOME
PROTEINS FOR THE GROWTH AND
VISION OF PEROXYSTUDIES OF
MULTIPLE ENDOCRINE BY O GENESIS,
WHILE LATER INFECTION THERE'S
INCREASED ABUNDANCE OF
PEROXISOME BIOGENESIS, SO THIS
CHANGE IN STRUCTURE YOU SEE
HERE, IT ACTIVATES THE SURFACE
OF THE PEROXISOME AND THE
PROTEINS THERE SUCH AS G-IMPACT
THAT ARE CONTROLLING THE
SYNTHESIS OF THESE LIPIDS THAT
WE THEN SHOW TO BE IMPORTANT FOR
THE SECONDARY ENVELOPEMENT SO
THEY ARE INTEGRATED INSIDE THE
VIRIONS AND THEY ARE CRITICAL
FOR MAKING INFECTIOUS VIRAL
PARTICLES AND IN AGREEMENT WITH
THIS WE TOOK PATIENT WHO IS HAVE
PEROXISOME DISORDERS AND FOUND
THEY ARE RESISTANT TO INFECTION,
SO THESE VIRUSES NEED THIS
METABOLIC FUNCTION MUCH
PEROXISOMES THAT ARE INTEGRATED
IN THE VERRIONS SO THIS WAS
UNEXPECTED AT THE TIME AND WAS
IT A NEW OUTLOOK TO FUNCTIONS
DURING INFECTION BECAUSE FOR
OVER 10 YEARS WE KNEW THAT
PEROXISOMES ARE IMPORTANT IN
INNATE IMMUNE RESPONSES THEY
HAVE MAPS AND ARE INVOLVE INDEED
HOST DEFENSE IN INFECTION BUT
NOW WE FOWBD THESE VIRUSES HAVE
ACQUIRED MECHANISM TO TURN THIS
INTO ANTIVIRAL ORGANELLE INTO A
PROFILE ORGANELLE BY REGULATING
LIPID SYNTHESIS, SO ALTOGETHER
SHE'S STUDIES HAVE PAINTED FOR
US A PICTURE OF THESE DYNAMIC
STRUCTURURE FUNCTIONS
RELATIONSHIPS AND WE WONDERED A
FEW THINGS LIKE 1 CAN WE START
MORE AND MORE TO UNDERSTAND WHAT
IS THE FUNCTION, WHAT ARE THE
FUNCTIONS OF THESE DIFFERENT
STRUCTURES, HOW ARE A DRIVEN AND
WHAT ARE THEIR FUNCTIONS BUT
ALSO, ARE THEY--ARE THEY
COORDINATED EVENTS.
ARE THESE ORGANELLE REMODELING
EVENTS ACTUALLY WORKING TO
INTRACELLULAR COMMUNICATION AND
EVEN INTERCELLULAR
COMMUNICATION.
SO INDEED, THIS IS NOT PUBLISHED
WHAT I WILL SHOW YOU NEXT BUT
INDEED WE KNOW THAT ORGANELLES
DON'T FUNCTION AS SEPARATE
ENDITYS BUT THEY FORM THESE
NETWORKS AND THIS IS
ACCOMPLISHED BY MEMBRANE CONTEXT
SITE PROTEINS THAT BRING
ORGANELLES THAT TETHER
ORGANELLES IN CLOSE PROXIMITY
LIKE 10-30 NANO MEETS AND BY
DOING THIS IT ALLOWS FOR
TRANSFER OF BIOMOLECULES THAT,A
LOWS THE RECRUITMENT OF
DIFFERENT PROTEINS THAT ARE ALSO
REGULATE THE FUNCTIONS OF THESE
ORGANELLE-ORGANELLE CONTACTS AND
HERE'S AN EXAMPLE OF 1 OF OUR
MOVIES WHERE IN [INDISCERNIBLE]
YOU SEE THE [INDISCERNIBLE] AND
THEY SHOW THE MITOCHONDRIA AND
THEY SHOW THE INTERACTIONS
BETWEEN THESE ORGANELLES SO WE
WONDER HOW DO VIRAL INFECTIONS
CHANGE OR REGULATE PROTEINS
DURING INFECTION AND WE PRO
DICTED THAT MANY OF THE CHANGES
THAT WE SEE IN THESE ORGANELLE
STRUCTURE AND FUNCTION HAVE TO
RELY ON THE REGULATION OF THESE
MEMBRANE CONTEXT SIDE PROTEINS
BUT WHAT WAS SURPRISING IS THAT
MCS HAS NOT BEEN THE FOCUS OF
STUDY DURING VIRAL INFECTION AND
THIS IS PERHAPS, ALSO DRIVEN IN
PART BY THE FACT THAT THESE ARE
NOT EASY PROTEINS TO STUDY, THEY
ARE MEMBRANE BOUND PROTEINS,
MANY OF THEM ARE LOW ABUNDANCE
PROTEINS, THEY ARE NOT--THERE
ARE NO ANTIBODIES AVAILABLE FOR
MANY OF THESE TO STUDY SO THIS
PROBABLY HAS DRIVEN SOME OF THIS
LACK OF KNOWLEDGE AND
UNDERSTANDING THIS.
ALSO THE METHOD OF CHOICE FOR
STUDYING CONTEXT SITES AND
ORGANELLE CONTACT USUALLY
PROVIDES THE OPPORTUNITY TO
STUDY 1 TACT AT A TIME OR JUST A
FEW SO WE ASKED CAN WE
UNDERSTAND HOW THESE ORGANELLE
REMODELING EVENTS ARE GLOBALLY
REGULATED AND WHETHER NMCS
PROTEINS PLAY A ROLE DURING
VIRAL INFECTION AND THE WORK HAS
BEEN DRIIVE BY THE SUPER
TALENTED Ph.D. STUDENT WHO
WORKED TOGETHER WITH HELEN AND
[INDISCERNIBLE] IN MY LAB.
SO IN ORDER TO DESIGN AN ASSAY
TO MONITOR MCS PROTEINS MASS
SPECTROMETRY IS IN A WAY IDEALLY
SUITED FOR THAT AND THE REASON
NEAR IS THAT THIS IS 1 OF THOSE
CASES WHERE WE KNOW THAT PROTEIN
ABUNDANCE IS DIRECTLY CONNECTED
TO THE EXTENT OF CONTACT SO IN
OTHER WORDS, THE LEVEL OF MCS IS
REALLY CRITICAL FOR THE AMOUNT
OF CONTACT BETWEEN ORGANELLES,
SO THIS IS SHOWN HERE, LET'S SAY
THE KNOCK DOWN OF THIS PROTEIN,
PPR59P 51, WHEN YOU SEE HOW THAT
IS BEING IMO TD DOWN YOU CAN SEE
HOW IT AAFFECTS THE
ER-MITOCHONDRIA.
WHEN IT EXPANDS IT ENHANCES THE
FACES OF THE INTERACTION BETWEEN
MITOCHONDRIA AND ER.
SO WE DECIDE TO TARGET A MASS
SPECT ROMET RADIOY APPROACH THAT
IS BASED ON PARALLEL REACTION
MONITORING TO TRY TO DETECT THE
PROTEINS IN HUMAN CELLS AND THIS
WAS QUITE A LENGTHY PROCESS SO
CAITLIN HAD TO MANUELLY CURATE A
LIBRARY OF FUNCTIONALLY
CHARACTERIZED MCS PROTEIN SO WE
TOOK INTO ACCOUNT THEIR CELL
TYPE SPECIFICITY, WE TOOK INTO
ACCOUNT TETHERS AND INTERACTORS.
AND THEN SHE HAD TO DESIGN A
LIBRARY OF PARAMETERS THAT WOULD
ALLOW US TO SPECIFICALLY DETECT
ONLY THOSE PARTICULAR PROTEINS
BY MASS SPECTROMETRY SO SHE FIND
THE PARAMETERS AND UNIQUE
PEPTIDES FOR THEIR DETECTION AND
THOSE SPECTRA LIBRARY FOR ALL
MCS PROTEINS.
THIS IS WAY EASIER THAN DONE,
THIS IS QUITE A LENGTHY PROCESS
BUT WOONS WE HAD THIS WE COULD
APPLY IT NOW TO MEASURE
ABUNDANCES IN 1 GOAL
SIMULTANEOUSLY ACROSS DIFFERENT
TIME POINTS AND CONDITIONS AND
USE THE INFORMATION TO INFER
WHETHER NEW CONTACTS OR CHANGE
CONTACTS BETWEEN ORGANELLES ARE
OCCURRING SO OUR ASSAY SO FAR
WHAT IT CAN DO IS SIMULTANEOUSLY
DETECT 90% OF THE KNOWN MCSs
AND THAT CONTAINS ALL MAJOR
ORGANELLE INTERFACES INCLUDING
PEROXISOME LIPID DROPMENTS AND
ALL FUNCTIONS THAT ARE MEDIATED
BY MCSs.
SO ONCE WE HAD THE ASSAY WE OOH
PLIED IT TO THE 4 VIRUSES I
MENTIONED THESE 4 ENVELOPE
VIRUSES WE STUDY IN THE LAB,
ATMB, HSV1, AND THE INFLUENZA A
AND HCOV-BET - CORONA VIRUS, SO
EVERY COLUMN REP ARE'S SENTS AN
MCS PROTEIN AND THEY ARE
CLUSTERED ACCORDING TO THE
ORGANELLE CONTACTS THAT THEY
FACILITATE AND THESE ARE EXAMPLE
ARE ER MITOCHONDRIA, THESE ARE
ER-GOLGI AND SO ON AND EVERY ROW
REPRESENTS A DIFFERENT TIME
POINT OF INFECTION AND WHAT YOU
CAN HOPEFULLY SEE, THE COLOR
REPRESENTS CHANGES IN PROTEIN
ABUNDANCE DURING--COMPARED TO
INFECTED CELLS.
SO WHAT CAN YOU MADLY SEE IS
THAT ATMB INFECTION TRIGGERS
GLOBAL UPREGULATION IN MCS
PROTEIN ABUNDANCES AND WE HAVE
AOBSERVED THIS AFTER INFECTION
WITH THE LABORATORY STRAIN OF
HCMV, AS WELL AS LOW PASSAGE
STRAIN THAT HAS MORE CLINICAL
RELEVANCE.
THIS IS QUITE IN CONTRAST WITH
CHANGES THAT HAPPEN DURING HSV 1
INFECTION, INFLUENCE OR OC43.
SO HSV 1 TRIGGERS MORE SUBTLE
CHANGES BUT ALSO MORE DISCREET
CHAIMPLES SO TO REGULATE CERTAIN
MCS PROTEINS IN THE SPECIFIC
MANNER.
AND OF COURSE THESE VIRUSES HAVE
REPLICATION CYCLES THAT ARE MORE
MATURE THAN THE CYCLE OF HCMV,
SO WHAT CAN WE LEARN FROM THIS,
WELL, AND THE FUNCTIONS THAT I
TOLD YOU EARLIER, SOMETHING THAT
WAS QUITE EVIDENT FROM OUR STUDY
WAS THAT THE PROTEIN TEGHT ERS
TO FACILITATE THESE, THESE 4
PROTEINS LISTED HERE, THEY WERE
ALL INCREASED IN ABUNDANCE
DURING HCMV AND THAT MADE A LOT
OF SENSE TO US BECAUSE IT'S
KNOWN THAT THE ER CONTACT IS
CRITICAL FOR SYNTHESIS AND THAT
IS INITIATING THAT PEROXISOME
AND IT WILL BE CONTINUED IN THE
ER AND IT'S ALSO IMPORTANT FOR
PEROXISOME GROWTH.
SO WE ASKED WHETHER THE INCREASE
THRA WE SEE IN PROTEIN CONTEXT
HERE IN THEIR ABUNDANCE
REPRESENT AN INCREASE IN ER,
EROXISOME CONTACT AND IF IT
HAPPENS DURING INFECTION.
AND THAT WAS THE CASE.
SO HERE IN UNINFECTED CELLS YOU
CAN SEE THE ROUND CELL THAT I
SHOWED YOU EARLIER ARE QUITE
DYNAMIC AND THEY TRACK ALONG
THESE ER TUBULES SHOWN HERE IN
CYAN, HOWEVER AFTER INFECTION
THEY BECOME MUCH MORE STABLY,
QUITE LARGE PEROXISOMES AND THEY
BECOME MUCH MORE STABLY ANCHORED
ON THE ER MEMBRANE AS SHOWN
HERE.
AND WE WERE ABLE TO ALSO PERFORM
REALLY--3D RECONSTRUCTION AND
SHOW HOW PEROXISOMES HAVE
INCREASED CONTACTS WITH ER
MEMBRANES AND STABLING AS THE
INFECTION PROGRESSES, SO WHAT'S
THE MEANING OF THIS AND WHAT IS
THE FUNCTION AND DOES THIS
TETHER, IS THE TETHERING
IMPORTANT FOR THE STRUCTURAL
CHANGES I MENTIONED EARLIER WITH
THE CURVED PEROXISOMES OR THE
INCREASE IN THE NUMBERS.
SO THE MAIN 2 TETHERS HERE ARE A
CBD5 AND THEN THE ER FIRST OF
ALL AS ECTOMYOSIN EXPECTED WE
COMPARE PEROX ISOMES AND THIS IS
JUST FOR INTEREST OF TAME SHOWN
AT LATE TIME POINT OF INFECTION
WE SEE THAT INCREASING MEMBRANE
AREA AND INCREASING PEROXISOME
NUMBERS.
WHEN WE IMOWK DOWN ACDB 5 WE
FIND THAT THE PEROXISOME NUMBERS
ARE INCREASED SO SOMEHOW THIS IS
A NEGATIVE FEEDBACK GROUP THAT
WAS NOT PREVIOUSLY KNOWN BUT THE
ABILITY OF THESE PEROXISOMES TO
BECOMING LARGE IS LOST DURING
INFECTION AND INDEED IF WE LOOK
AT ACDB 5 LOCALIZATION WE FIND
IT LOCALIZES SPECIFICALLY TO
ENLARGE THESE TO INFECTION, IF
YOU DO THE OPPOSITE EXPERIMENT
AND WE OVEREXPRESS THIS TETHERED
PROTEIN TO ICDB5, WE FIND THAT
THE NUMBER OF PEROXISOME IS
DECREASED BUT THE SIZE IS
INCREASED, IT FORMS THE LARGE
PEROXISOM, SOW THIS IS REQUIRED
BUYER THE PROVIRAL CHANGES TO
THE SIZES THAT I SHOWED YOU
EARLIER.
SO WE ALSO SHOWED THIS IS INDEED
CONTRIBUTES TO THE VIRAL TITERS
CHANGES WE SHOWED, WE REPORTED
EARLIER FOR HCMB, AND WE FOUND
THE SAME IS TRUE FOR INFLU
FLUENZA A AND THE BETA CORONA
VIRUS HCOV-O C43 SO THEY ALL
NEEZ THE CONTACT IN ORDER TO
CONTROL THEIR FUNCTIONS BUT I
WANT TO SPEND KIND OF THE
REMAINING FEW MINUTES I HAVE
HERE ON SOMETHING NEW THAT WE
DISCOVER THAT IS QUITE
INTRIGUING AND IT'S BASED ON
THIS OBSERVATION WE HAD ON ER
MITOCHONDRIAL CONTACTS SO TO
START BY PUTTING THIS IN BROADER
CONTACTS WE HAVE BEEN INTERESTED
IN THE REGULATION FOR A WHILE
DURING INFECTION, AND IT'S VERY
WELL KNOWN THAT MANY VIRUSES ARE
CONTROLLING MITOCHONDRIAL
FRAGMENTATION AND INFUSION
DURING INFECTION IN ORDER TO
CONTROL APOPTOSIS, IMMUNE
SIGNALING AND ATP PRODUCTION.
AND USUALLY INCREASED
FRAGMENTATION THAT IS DRIVEN BY
MANY VIRUSES IS CONNECTED WITH
REDUCED ATP PRODUCTION, SO
REDUCED BIOENERG JETTIC OUTPUT,
HOWEVER HCMB INFECTION KIND OF
PRESUNTS A CONUNDRUM, A PARADOX
BECAUSE MITOCHONDRIA IS
FRAGMENTED AFTER INFECTION WITH
HCMB, HOWEVER SOMEHOW THEY ARE
BIOLOGICALLY ENHANCED AND THE
MECHANISMS REMAIN MYSTERIOUS AND
SOMETHING WE NOTICED FROM OUR
MCSN, ANDA PROTEINS ARE INNED AS
THE INFECTIONS DONES AND THE
TIME WHEN WE SEE AND ALSO WHEN
WE SEE MANY CELLULAR CHANGES IN
CELLULAR METABOLISM, INCREASED
FLUX THROUGH GIVING--YOU COLSIS,
INCREASED PHOSPHORYLATION, SO WE
WONDERED WHETHER THESE CHANGES
IN ER MITOCHONDRIAL CONTEXT,
ACTUALLY SOMEHOW DRIVING THIS
INCREASED--THIS INTERESTING
PHENOTYPE THAT WE CAN'T EXPLAIN.
BUT FIRST IN ORDER TO LOOK AT
THIS, WE HAVE TO CONVINCE
OURSELVES THAT CHANGES WE SEE
HERE IN PROTEIN CONTACT, IN
PROTEIN ABUNDANCES, ACTUALLY
REFLECTS THE CHANGE IN
ORGANELLE-ORGANELLE TON TACT.
SO USING FLUORESCENT MICROSCOPY
AND [INDISCERNIBLE] MICROSCOPY,
WE MONITORED THE INTERACTION
BETWEEN MITOCHONDRIA AND ER, AND
WE NOTICED THIS INTERACTION THAT
HAS BEEN PREPORTED MANY TIMES
BEFORE IN WHICH THESE ER TUBULES
ARE WEAVING ACROSS THE
MITOCHONDRIA AND YOU CAN SEE
THESE LITTLE CONSTRICTION SITES
WHERE THE--ON MITOCHONDRIA WHERE
THE MEMBRANE TUBULES ARE
CROSSING.
AND THIS IS A DYNAMIC
INTERACTION AS YOU SEE HERE,
HOWEVER, AFTER INFECTION, THE
INTERACTION CHANGES QUITE
STRIKINGLY, THE MITOCHONDRIA
BECOME ENCAPSULATED AND STABLY
IN THE ISHT ACTION WITH THE
MEMBRANES AND WHEN WE ALSO
LABELED AND THE OUTER MIGHT ON
CHOND RIA MEMBRANE, WE CAN
CONFIRM THAT THE CONTACT BETWEEN
THE MITOCHONDRIA AND THE ER IS
INCREASED BUT ALSO THIS IS
ASYMMETRIC, IT HAPPENS ON 1 SIDE
OF THE MITOCHONDRIA WHERE THE
CONTACT IS INCREASED SO THIS IS
QUITE DIFFERENT THAN THE DYNAMIC
TUBULE CROSSINGS THAT ARE
USUALLY REPORTED FOR THESE
INTERACTIONS AND PRESENTS A NEW
MITOCHONDRIAL ER CONTEXT
STRUCTURE THAT HAS NOT BEEN
REPORTED BEFORE, SO WE USE SUPER
RESOLUTION MICROSCOPY AND USE IT
TO GAIN BETTER UNDERSTANDING OF
WHAT WE ARE LOOKING AT, INDEED
WE FOUND THAT THE ER
FORMS--INCAPSULATES THE
MITOCHONDRIA IN LITTLE POCKETS
AND THIS IS TABLE AND ASYMMETRIC
MEMBRANE CAPSULATION, SO WE TURN
THIS STRUCTURE MENCs FOR
MITOCHONDRIA-ER ENCAPSULATIONS
AND WE MONITOR DURING THE
PROGRESSION OF INFECTION AND WE
NOTE THAD AS THE INFECTION
PROGRESSES, THIS CONTACT,
MENCs BECOME THE PRIMARY
CONTACT TYPE FOR ER MITOCHONDRIA
AND WE ALSO NOTICE THAT AS THESE
MENCs ARE FORMED THIS HAVE
INCREASED IN THIS STABLE TYPE OF
INTERACTION BETWEEN THESE 2
ORGANELLES AND THE DYNAMIC
INTERACTION PREVIOUSLY NOTICED
IS DECREASING.
SO WE THEREFORE ASK WHAT ARE THE
ER MITOCHONDRIAL MCS PROTEINS
THAT ARE LOCALIZING TO THE
MENCs, AND WHAT IS THE
STRUCTURAL FORMATION.
SO WE LOOK AT THE MASS
SPECTROMETRY DATA ISSUES THE 2
THAT ARE BETWEEN THESE CONTACT
WERE THE TETHERING COMPLEX THAT
IS FORMED BY PTP151 AND VAP-B
AND THIS IS KNOWN TO BE IN
CA-FLUX, AND BIOENERGETICS AND
AUTOPHAGY, SO WE LOOK AT THESE
PARTNERS AS THE INFECTION
PROGRESSES AND THAT ALLOWS US TO
DISCOVER THE FORMALITY OF THIS
AND WE FOUND MENCs ARE FORMED
AND ABOUT 48 HOURS POST
INFECTION, THEN PTPIP51 IS AT 72
HOURS POST INFECTION AND BOTH OF
THESE PROTEINS, VAP-B AND
PTPIP51 ARE LOCALIZATIONOT 1
SIDE OF THE MITOCHONDRIA AND WE
FURTHER VALIDATED THIS
INTERACTION USING PROXIMITY
LIGATION ASSAY TO CONFIRM THE
INTERACTION AND ALSO OBSERVE THE
INTERACTION FOR THE ENDOGENOUS
PROTEINS AND WE COULD SEE HOW
LATENT INFECTION THIS ACTUALLY
HAPPENS.
SO SINCE PTPIP51 WAS THE LAST 1
RECRUITED TO THIS, SO MENCs
FORM AND THEN VAP-B AND THEN
PTPIP51 AND WE ASKED THE
FUNCTION OF THIS IS TO USE THIS
WE USE THE ASSAY WE DEVELOPED
RECENTLY IN THE LAB AND THIS WAS
DEVELOPED BY MICHELLE KENNEDY A
VERY TALENTED STUDENT IN THE
LAB.
AS I TOLD YOU EARLIER THESE
VIRUSES ISSUES HERPES VIRUSES
ARE REALLY LARGE, THEY HAVE
LARGE GENOMES WE LACK ANTIBODIES
FOR MOST OF THESE VIRAL PROTEINS
SO DETECTING THEM AND
QUANTIFYING THEM AND ACCURATELY
DURING INFECTION IS NOT TRIVIAL
SO MICHELLE HAS DEVELOPED THIS
TARGETED MASS SPECTROMETRY ASSAY
TO DETECT THE MAJORITY OF THE
PROI DON'T MEANS OF THESE
VIRUSES, HCMV, AND KSHV AND SHE
SHOWED THAT SHE CAN CAPTURE WILL
TEMPORAL EXPRESSION OF VIRAL
GENE EXPRESSION, SHE CAN
PROJECTION MONITOR
THE TEMPORAL DRUGS OR SMALL MOLL
COWLS OR GENETIC PROTURBATIONS
AND TOGETHER WITH TOM KRISTI WE
SHOWED THAT WE,A MRI THIS NOT
ONLY TO CELL CULTURE BUT ALSO
ANIMAL MODELS OF INFECTION, SO
THEREFORE WE USED OUR HCMV
VERSION OF THIS,A SAY, BUT THIS
TIME TO OUR CONDITION OF PTPIP51
KNOCK DOWN TO FIND OUT THE
FUNCTIONAL CONSEQUENCE OF THIS
RECRUITMENT OF PTPIP51.
SO WE FOUND THE CONTROL KNOCK
DOWN AND THE PTPIP51 KNOCK DOWN
AND YOU ARE LOOKING AT FEW VIRAL
PROTEINS ILLUSTRATED HERE AND WE
FOUND THAT THIS--THE KNOCK DOWN
OF PTPIP51 DOES NOT IMPACT THE
EXPRESSION OF EARLY VIRAL GENES
BUT RATHER DECREASES THE LEVELS
OF DELAYED EARLY AND DELAYED
VIRAL PROTEINS SO PROTEINS THAT
SYNTHESIZE AROUND 48 AND 72
HOURS POST INFECTION.
AND THIS DECREASE IN VIRAL
PROTEIN LEVELS INDEED LEADS TO
AN OVERHUNDRED FOLD DECREASE IN
VIRUS TITER AND THIS IMPACT ON
THE VIRUS TIGHTER IS MAINTAINED
OVER TIME.
SO ALTOGETHER WHAT WE NOW
UNDERSTAND IS THAT PROTEIN
PTPIP51 INCREASE IN ABUNDANCE IN
INFECTION AND ACCUMULATING
ASSIMETRICALLY THE MENCs AND
IT'S CONTECTING TO INFECTION.
SO I JUST TOLD YOU PTPIP51 IS
PROVIRAL, SUPPORTING VIRAL
PRODUCTION BUT TURNS OUT THAT
THE TEMPERRALLITY IS ABSOLUTELY
KEY AND THIS IS SOMETHING
SURPRISING AT THE TIME AND MADE
SENSE TO US SO IF WE PREMATURELY
INCREASE ER MITOCHONDRIAL
CONTACT IT TURNS OUT THIS IS
ANTIVIRAL, IT'S NOT PROVIRAL, IF
WE SET THIS INTERACTION TOO
EARLY IT'S NOT DOING WHAT WE
THOUGHT IT WOULD DO SO FOR
EXAMPLE, THIS BRINGS TOGETHER
EARLIER INFECTION AND WE SEE
CONCENTRATION DEPENDENT AND
DECREASE VIRUS PROTUKS, SO FEW
QUESTION, FIRST OF ALL WHY IS IT
PROVIRAL ONLY LATENT INFECTION,
ONLY AFTER 48 HOURS WHEN THIS
CONTACT, THE FORMATION OF THIS
CONTACT IS ONLY PROVIRAL LATENT
INFECTION AND WE THINK THAT IT'S
BECAUSE IT NEEDS THE WRITE
CONTEXT, IT NEEDS THE
MITOCHONDRIAL FRAGMENTATION TO
OCCUR CAN AND IT NEEDS THE
LOCALIZATION OF THE FACTORS TO
THE MITOCHONDRIA.
SO FROM THE PATTIAL PROTEOMICS
STUDY I MENTIONED EARLIER, I
TOLD YOU WE FOUND OUT WHICH 1S
LOCALIZE TO PRODUCE ORGANELLES
TO INDUCE THESE CHANGES AND WE
FOUND THIS IS THE WORK OF
CORA BETSINGER IN MY LAB, SHE
PUBLISHED THIS LAST YEAR, BUT WE
FOUND OUT THAT THIS LOCALIZED TO
THE MITOCHONDRIA AND BY USING
LIVE CELL METABOLIC ASSAYS, SEA
HORSE ASSAY, SHE SHOWED THAT
VIRAL PROTEIN IS NECESSARY AND
SUFFICIENT TO INCREASE
PHOSPHORYLATION AND WE THINK
THIS IS ACTUALLY DOWN STREAM OF
THE MAIN FORMATION WHEN PUL13 IS
LOCALIZED THERE.
AND SHE ASKED HOW IS THIS
ACCOMPLISHING THIS AND TO DO
THIS SHE ANALYZE THE TEMPORAL
INTERACTIONS, PROTEIN
INTERFERON-GAMMA T-CELL ACTIONS
AS THE PROTEIN PROGRESSES AND
SHE USED THE PROGRAM COMPUTATION
PLATFORM THAT WE DEVELOPED A FEW
YEARS AGO CALLED INTERVISTA, AND
SHE INFECTED THESE CELLS WITH
THE PROTEIN, PERFORMED AN
AFFINITY PURIFICATION DURING
DIFFERENT TIMES OF INFECTION AND
USED THIS TO VISUALIZE THESE
INTERACTIONS AND SHE FOUND THAT
48 HOURS POST INFECTION, WHEN WE
SEE MENC FORMATION, UL13 GOES
THROUGH THE REMODELING
COMPLEXES, AND USING SUPER
RESOLUTION MICROSCOPY, PUL13 IS
SUFFICIENT TO CHANGE THE
ARCHITECTURE OF THE CRISTACE
STRUCTURE AND THE SURFACE AND
VOLUME OF THIS.
SO THIS REPRESENTS THE FIRST
VIRAL PROTEIN THAT CO MODULATE
THAT CAN LOCALIDES AND THE
STRUCTURE OF MITOCHONDRIA BY
ITSELF TO IN ORDER TO INCREASE
BIOINNER GENETICS.
SO I TOLD YOU THAT THIS POST
VIRAL FUNCTION HAPPENINGS LATE
IN INFECTION AND WHY WE THINK IT
HAPPENS BUT LASTLY IF WE COME
BACK TO THIS IDEA THAT IF WE
TETHER THIS EARLIER, SUDDENLY
THIS BECOMES ANTIVIRAL, THE
QUESTION IS WHY DOES THAT
HAPPEN.
AND WELL, IN USING THE SYNTHETIC
TETHER WE FOUND THAT IF
ER-MITOCHONDRIA ARE FORCED TO
INTERACTION TO EARLY AND
MAINTAIN AS A STABLE INTERACTION
TOO EARLY, IT ACTIVATES STING
AND THEREBY ACTIVATES THE TTK,
AND SIGNALING KASP A R CASE AND
INDUCES CYTOKINE EXPRESSION.
SO IN AGREEMENT WITH THE
ACTIVATION OF THIS BROAD HOST
DEFENSE MECHANISM, WE SEE THE
TETHER FORCING THIS INTERACTION
EARLY ON, ALSO DECREASES THE
VIRUS TITER FOR HSV 1 AND
INFLUENZA A IN A CONCENTRATION
DEPENDENT MANNER FOR THIS DATA
AND THIS IS PERHAPS WHY FOR
THESE VIRUSES THAT HAVE MUCH
SHORTER REPLICATION CYCLES WE
SEE THAT EARLY IN INFECTION THEY
IMMEDIATELY START TO DOWN
REGULATE THE ER-MITOCHONDRIAIAL
CONTACT PROTEIN OTHERWISE THEY
WOULD FORCE THIS TO HAPPEN WHEN
INDUCED SIGNALING.
OKAY, SO WITH THAT I TOLD YOU
HOW THESE ARE TEMPORALLY
RESTRUCTURED AND HOW THE
TEMPORALITY OF THE MCS
REGULATION IS CRITICAL AND HOW
WE HAVE DISCOVERED THIS NEW
STRUCTURE OF MITOCHONDRIA ER
ENCAPSULATION THAT WE
DEMONSTRATE THAD IS NECESSARY
FOR VIRUS PRODUCTION AND IT'S A
PRO VIRAL.
AND HOW THIS REMODEL SUGGEST
QUITE BROADER IT'S QUITE GLOBAL
DURING ATV INFECTION AND WE
DEVELOP THIS ASSAY TOO BE ABLE
TO MONITOR ALL THESE MCSs IN
THE SAME TIME AND HOW--THIS
ALLOWED US TO FIND VIRUS
SPECIFIC AS WELL AS SHARED MCS
WIRING EVENT SUCH AS
ER-PEROXISOME INN TACT.--CONTAC.
SO WITH THAT I WILL THANK THE
PEOPLE I HIGHLIGHTED TODAY, THE
COLLAB RETIREDDORS, FUNDING AND
MY PHENOMENAL LAB AND I WANT TO
THANK YOU IF ARE YOUR ATTENTION
AND AGAIN THE HONOR FOR INVITING
ME HERE TODAY.
[ APPLAUSE ]
>> OKAY, THANK YOU.
ANYONE'S FREE TO USE THE MICs
IN
IN THE AISLES.
I DO MY BEST TO READ THEM FOR
YOU.
THIS CAME IN EARLY IN YOUR
LECTURE.
>> SURE.
>> BASICALLY THE--THIS IS FROM
SOMEONE HERE, [INDISCERNIBLE],
BASICALLY THE TEMPERATURE IS
CHANGED WITH AIR COMSUMPTION
ESPECIALLY HIGH TEMPERATURE, DID
YOU CHANGE TD CULTURE CONDITION
ESPECIALLY OXYGEN AND CO-2
COMSUMPTION?
>> OKAY, SO THIS PROBABLY REFERS
TO THE THERMOCOAGULATION
PROFILES WHERE THE PRINCIPLE IS
THAT YOU'RE DIN'RE INDUCE
THANKSGIVING AG DPREIGATION OF
THE PROTEIN BECAUSE YOU'RE
INCREASING THE TEMPERATURE.
OF COURSE AT THAT TIME YOU'RE
JUST SIMPLY AGGREGATING THE
PROTEIN, YOU'RE NOT TRYING TO
PRESERVE THE CELL FUNCTIONS IN A
CELL IT'S MAINLY TO AGGREGATE
THE PROTEIN AND SEE WHICH
PROTEIN-PROTEIN INTERACTIONS BUT
SOMETHING THAT'S VERY NICE
QUESTION BECAUSE SOMETHING THAT
WE HAVE ACTUALLY DONE IS NOW TO
REVISIT A FEW THINGS FOR THAT
WORK FLOW IN PARTICULAR THE
TEMPERATURES THAT ARE NEED IN
ORDER TO GET THOSE CURVES AND WE
FIND THAT YOU DO NOT NEED TO GO
THAT HIGH IN TEMRATURE IN
ORDER TO BE IN THE SWEET SPOT
WHERE YOU BEST PREDICT
COAGULATION PROFILES AND ALSO
MANY PROTEINS AS YOU PROBABLY
GUESS THIS IS COMPUTATIONALLY
INTENSIVE, RIGHT?
SO MANY PROTEINS DO NOT BEHAVE
VERY WELL AND THEY ARE HAVING
CURVES IN ALL DIFFERENT DREKS
AND AGGREGATING PROTEINS OR
MEMBRANE BOUND PROTEINS DO NOT
BEHAVE WELL, SO TRAVIS IN MY LAB
HAS LICENSED BUFFER CONDITIONS
TO TRY TO CAPTURE MORE AND MORE
OF THE PROTEOME BUT HE ALSO HAS
TESTED EXPERIMENTALLY AND ALSO
COMPUTATIONALLY MONITORS WHAT
ARE THE TEMPERATURES SO WE NEED
ONLY 5 TEMPERATURES IN ORDER TO
GET REALLY GREAT INFORMATION
ABOUT AGGREGATION CURVES AND
THIS BRINGS IT INTO DIFFERENT
TEMPERATURES IN TERMS OF
MULTIPLEXING THIS AND MAKING IT
MUCH FASTER.
BUT HE ALSO DEVELOPED THEE
MACHINE ALGORITHMS FOR BETTER
ANALYZING THE DATA.
YES.
SNRS I HAVE 2 QUESTIONS, 1 IS TO
THE LAMINA ACETYLATION, DO YOU
KNOW THE ENZYME THAT DOES IT?
IS IT A SPECIFIC ACETYLATION OR?
>> SO WE ONLY VERY LITTLE TABLED
FOR THE MOMENT.
SO I CAN TELL YOU WHAT WE KNOW
SO FAR, SO IT SEEMS LIKE P300
ARE THE STILL TRANSFER ACE THAT
DOES THAT, FOR THE
DEACETYLATION, WE TESTED WITH
HBACK INHIBITORS AND WE STARTED
TO LOOK AT CERTUEINS 1 AND 6 AND
WE NOTICE THAD 6 IS GOING TO THE
PERIPH RADIOY AND THEREFORE IS
KIND OF AT THE RIGHT TIME AND
PLACE TO TRY TO DEACETALATE THAT
BUT WE HAVE NOT YET LOOKED INTO
THAT TO VALIDATE IT.
>> BECAUSE THIS COULD BE MAB
EPIGENETICCULATED
THERAPEUTICALLY.
>> YES, YES, AND AS WE ALL KNOW
THEY ARE RESTING THE CELL SITE,
SO THEY ARE REPLICATING THE THIS
G1 S SO IT'S PERFECT FOR THE
HOST MACHINERY TO BE IMPRESSED
BUT NOT FOR HOST REPLICATION TOO
HAPPEN, RIGHT?
SO IT MAKES SENSE THAT THE
ACETYLATION THAT WE SEE IT IN
UNINFECTED CELLS IS THE SAME,
AND THE SECOND QUESTION IS TO
TETHERING, THE LAST TOPIC, DO
YOU THINK IT'S MORE GENERAL
PHENOMENON OR IT CAN HAPPEN IN
ALL INFECTIONS OR EVEN WHATEVER
PERTURBATIONS THE CELLS ARE AT
AND AND IF SO, BECAUSE IT CAN--I
WAS THINKING IT CAN AFFECT OTHER
KINDS OF EXPERIMENTS SO FOR
EXAMPLE CELLULAR LOCALIZATION
EXPERIMENTS CAN BE CHANGED
BECAUSE OF THAT AND NOT BECAUSE
THE PROTEIN ACTUALLY CHANGED THE
LOCALIZATION, THEY CAN LOOK LIKE
THAT.
>> SO I THINK IT MAKES SO MUCH
SENSE AND I'LL COME BACK TO THE
PARTICULAR TETHERING AND TELL
YOU WHY I THINK IT MAY OR MAY
NOT BE BUT IT MAKES SO MUCH
SENSE FOR MEMBRANE CONTEXT SITE
PROTEINS TO BE REGULATED DURING
INFECTION BECAUSE IF WE THINK
ABOUT IT, IT'S SO CRITICAL
PROTEINS FOR INTRACELLULAR
COMMUNICATION FOR THESE VERY
FAST SIGNAL THAGOREAN WE SEE IN
RESPONSES.
SO WHEN WE LOOK AT ALL THOSE
STRUCTURAL CHANGES, IT'S
IMPOSSIBLE FOR THE EARTH
STRUCTURE LIKE THAT NOT TO
IMMEDIATELY INFLUENCE ITS
ENVIRONMENT AND ALSO COMMUNICATE
THAT, AND WHAT BETTER WAY THAN
DIRECT CONTEXT BETWEEN
ORGANELLES THAT CAN QUICKLY MOVE
SMALL MOLECULES OR RECRUIT
PROTEINS.
I'VE BEEN FOR A WHILE FASCINATED
WITH THE MECHANO RESPONSES AND
THAT HAPPEN DURING INFECTION AND
THOSE ARE CLEARLY INVOLVED HERE
AS WELL.
NOW COMING BACK TO THE TETHER,
DEPENDING ON WHICH, SO FOR THE
MENCs, FOR THESE STRUCTURES, I
ACTUALLY THINK THAT THIS IS
PROBABLY FAIRLY SPECIFIC, SO IF
WE JUST INDUCE MITOCHONDRIAL
FRAGMENTATION, OR IF WE PERTURB
THE ER FUNCTIONS WE DO NOT SEE
MENCs FORMING SO WE THINK THAT
THIS IS ACTUALLY HAVING TO DO
WITH THAT REALLY INTERESTING
PHENOTYPES OF MITOCHONDRIA BEING
FRAGMENTED BUT BEING STILL
BIOGENETICALLY ENHANCED SO WE
NOW SHOW AND HAVE PRELIMINARY
DATA THAT WE THINK THAT S&P
PROTECTING THE MITOCHONDRIA AND
PERHAPS DECREASING MIGHT O
PHAGEY AND THE WORK OF OTHERS SO
THIS IS THE WORK OF WILL AND
CAITLIN COOK SO I DO NOT THINK
THAT IT WILL BE RELEVANT TO ALL
VIRAL INFECTIONS BECAUSE OF
THIS, HOWEVER, WE NOW HAVE
EVIDENCE THAT IT'S RELEVANT IN
OTHER CIRCUMSTANCES.
WE THINK THAT IT'S RELEVANT IN
CERTAIN NEUROGERONTOLOGYSTS
GENRATIVE DISEASES BUT IT'S ALSO
RELEVANT IN CANCER, ISY SO WE
LOOK AT CANCER CELLS BECAUSE
THEY TRIGGER A SIMILAR CHANGE IN
CELLULAR METABOLISM LIKE HCMB SO
WE DO SEE THAT MENCs DO FORM
THERE.
>> SO THE MEMBRANE IS A
PHENOMENON, THANK YOU FOR THIS
INSPIRING TALK.
>> COUPLE MORE THIS, IS FROM
DR. [INDISCERNIBLE] AT THE NIH,
MIGHT BE RELATED TO THAT.
COULD THE I HAVE RUGS DNA
BINDING PROTEIN RECOGNIZE DNA
ENDS WITHOUT TELEOR MERES, DO
THE SAME PROTEINS RECOGNIZE
CIRCULAR VIRAL DNA,.
>> SO THAT'S REALLY INTERESTING
AND I DON'T KNOW, THIS IS
PROBABLY REFERRING TO THE DNA
SENSING STORY, THE NUCLEAR DNA
SENSING AND HOW--YES,--AND HOW
THESE HOST PROTEIN I516
RECOGNIZES VIRAL DNA INSTEAD OF
HOST DNA AND 1 THING I CAN SAY,
WE HAVE DONE THIS BUT THIS WAS
DONE INVITRO WHEN WE LOOKED AT
CIRCULAR DNA VERSUS LINEAR DNA,
WE WERE SPECULATING IT WOULD
ATTACHED TO LINEAR BECAUSE IT
WOULD DAMAGE DNA BUT WE FIND IT
BINDS TO BOTH CIRCULAR AND
LINEAR IN A SIMILAR FASHION, WE
HAVE SINCE THEN DONE OUR CHP-SEQ
AND ATTACH SEQ AND BYPASSEDS IN
A SEQUENCING MANNER AND IT'S
KNOWN ALREADY THAT IT'S--SO THE
PYRENE DOMAIN HAS ALL THIS BASIC
RESIDUES AND WE FOUND CERTAIN
RESIDUES THAT CAN BE MUTATED AND
WE FOUND HOW THOSE ARE
POLYMERIZED AND HOW THEY BIND TO
VIRAL DNA.
YEAH, THAT ARE SO MANY THINGS TO
SAY THERE, SO YOU CAN STOP ME
WHEN YOU WANT.
>> WELL MAYBE WE CAN SNEAK IN 1
MORE BEFORE WE CLOSE THIS, IS
FROM RON GERMANE AT THE NIH
HERE, YOU SHOWED THAT MENC FORM
EARLY BEFORE THE ADHESIVE
INTERACTIONS WITH THE PROTEINS
THAT CONTRIBUTE TO VIRUS
REPLICATION, WHAT ALLOWS
FORMATION OF MENC THAT INVOLVES
MEMBRANE TO MEMBRANE INTERACTION
OF ORGANELLES BUT NOT THE
MOLECULAR PAIR YOU MENTIONED AS
CRITICAL FOR ER MIGHT O CONTACT.
SO CAN YOU PLEASE REPEAT THAT
BECAUSE I MISSED THE BEGINNING
SO IT'S ABOUT THE MENCs,.
AND--SIT'S FINE, YES.
YES, OKAY, SO YOU SHOW THAT
MENC FORM EARLY BEFORE THE
ADHESIVE INTERACTION WITH THE
PROTEIN THAT CONTRIBUTES TO
VIRAL REPLICATION.
>> YES, SO IT'S FORM BIDE MANY
PROTEINS SO REMEMBER THE HEAT
MAP AND I SHOWED HOW MANY OF
THEM ARE KNOWN TO INFORM THAT
THE ER MITOCHONDRIAL CONTACT BUT
THEN THE PROTEINS, THE VAP-B, I
THINK THAT'S WHAT THE QUESTION
REFERS TO AND PTPIP51 ARE
RECRUITED THERE.
IF YOU LOOK AT PTPIP51 IT'S
DISTRIBUTED, DIFFUSED AND
RELOCALLIZED IT GOES TO THE
TETHERING WITH VAP-B APPROXIMATE
HOW IT'S LOCALIZED ON THE
MITOCHONDRIA, SO, AND THAT'S
KNOWN--THAT'S NOT KNOWN FOR
INFECTION OR NOT KNOWN THAT IT
HAPPENS IN A SYMMETRIC THING AND
THAT RELOCALLIZATION IS KNOWN TO
BE IMPORTANT FOR THE FORMATION
OF THIS TETHERING COMPLEX AND
THERE ARE OTHER PROTEINS THAT
EARLIER ON ARE BRING THESE 2
ORGANELLES IN CONTACT THAT ARE
INCREASE NOTHING ABUNDANCE AS I
SHOWED THERE.
>> MAYBE--CLOSE THIS OUT?
>> THANK YOU FOR ALL THOSE GREAT
QUESTIONS.
>> THANK YOU VERY MUCH ILEANA
FOR THE GREAT TALK, I AM SURE IT
WILL BE INSPIRING FOR A LOT
PEOPLE AND THANK YOU VERY MUCH
FOR COMING AND AND WONDERFUL
ATTENDANCE IN PERSON AND ONLINE.
THANK YOU SO MUCH.
THANK YOU AGAIN.
[APPLAUSE ]