R01AI198154

TAR RNA BINDING TO INI1/SMARCB1 AND ITS ROLE IN HIV-1 TRANSCRIPTION AND LATENCY REACTIVATION - ABSTRACT THE GOAL OF THIS APPLICATION IS TO STUDY THE ROLE OF INTERPLAY BETWEEN THE COMPONENTS OF CHROMATIN REMODELING SWI/SNF (BAF COMPLEX) AND HIV-1 TRANSCRIPTION MACHINERY, FOCUSING ON THE INTERACTION OF A BAF COMPONENT, INI1 (INTEGRASE INTERACTOR 1) WITH TAR RNA. HIV-1 RESERVOIRS ARE A MIXTURE OF LATENT CELLS HARBORING PROVIRUSES SILENCED AT TRANSCRIPTIONAL LEVEL. CURE STRATEGIES NEED A DEEPER UNDERSTANDING OF HIV-1 TRANSCRIPTIONAL REGULATION. HIV-1 TRANSCRIPTION, INITIATED BY RNA POL II, PAUSES PRODUCING SHORT TAR TRANSCRIPTS. PTEFB RECRUITMENT TO TAR BY TAT OVERCOMES THIS TRANSCRIPTIONAL PAUSE, FACILITATING ELONGATION. BEYOND TAT, THE ACTION OF CHROMATIN REMODELING COMPLEXES (CRCS) IS REQUIRED TO FACILITATE ELONGATION. THE BAF COMPLEXES CBAF AND PBAF PLAY DISTINCT ROLES. WHILE CBAF REPRESSES PROVIRAL TRANSCRIPTION BY MAINTAINING NUCLEOSOMES IN AN UNFAVORABLE STATE, PBAF REMODELS NUCLEOSOMES TO FACILITATE ELONGATION. INI1 IS A COMPONENT OF BOTH CBAF AND PBAF, AND ITS ROLE IN TRANSCRIPTION IS NOT FULLY UNDERSTOOD. INI1 WAS IDENTIFIED AS A BINDING PARTNER FOR HIV-1 INTEGRASE (IN) AND EXERTS MULTIFACTED ROLES IN VIRUS ASSEMBLY, PRODUCTION AND MORPHOGENESIS. INI1 HAS MULTIPLE FUNCTIONAL DOMAINS. IN BINDING RPT1 DOMAIN STRUCTURALLY MIMICS TAR RNA & IS NECESSARY FOR LATE EVENTS. WE HAVE MADE A NOVEL OBSERVATION THAT ANOTHER DOMAIN OF INI1, THE N-TERMINAL WINGED HELIX DNA BINDING DOMAIN (WHD) SPECIFICALLY BINDS TO TAR RNA AND THAT THIS INTERACTION IS NECESSARY FOR MEDIATING HIV-1 TRANSCRIPTIONAL ELONGATION. THESE EXCITING RESULTS SUGGEST THAT DIFFERENT FUNCTIONAL DOMAINS OF INI1(RPT1 AND WHD) INVOLVED IN “TAR RNA MIMICRY” OR “TAR RNA BINDING” REGULATE DISTINCT STAGES OF REPLICATION. WE HYPOTHESIZE THAT INI1 WHD DOMAIN-TAR INTERACTION IS NECESSARY FOR RECRUITMENT OF PBAF TO HIV-1 LTR FOR TRANSCRIPTIONAL ELONGATION AND LATENCY REACTIVATION. DISRUPTING THIS INTERACTION RESULTS IN TRANSCRIPTIONAL REPRESSION. WE WILL INVESTIGATE THE ROLE OF THIS NOVEL INI1:TAR RNA INTERACTION IN HIV-1 TRANSCRIPTION AND LATENCY REACTIVATION. THIS IS A MULTI-PI APPLICATION INVOLVING DRS. KALPANA (HIV-1 VIROLOGIST), HENG (NMR BIOPHYSICIST) AND ZOU (COMPUTATIONAL BIOLOGIST/PROTEIN-RNA STRUCTURE). IN AIM 1, WE WILL CHARACTERIZE INI1-WHD:TAR INTERACTION IN VITRO AND IN VIVO VIA MOLECULAR/GENETIC ANALYSES (KALPANA/HENG). WE WILL EMPLOY ALANINE SCANNING MUTAGENESIS BASED ON WHD NMR STRUCTURE TO TEST WHD:TAR INTERACTION. WE WILL USE BIOPHYSICAL & BIOCHEMICAL APPROACHES TO PROBE TAR STRUCTURAL ELEMENTS REQUIRED FOR THIS INTERACTION. IN AIM 2, WE WILL EMPLOY COMPUTATIONAL MODELING AND NMR TO DETERMINE THE STRUCTURE OF INI1- WHD:TAR RNA COMPLEX (ZOU/HENG). IN AIM 3, WE WILL DETERMINE THE ROLE OF INI1:TAR INTERACTIONS IN HIV-1 TRANSCRIPTION, LATENCY REACTIVATION AND MECHANISM OF ACTION (KALPANA). WE WILL ANALYZE THE EFFECT OF TAR- INTERACTION-DEFECTIVE (TID) INI1 MUTANTS ON TRANSCRIPTION OF LTR-REPORTERS AND FULL-LENGTH HIV IN INI1-/- CELLS. LATENT CELLS IN WHICH TID-INI1 MUTANTS ARE KNOCKED IN (KI) WILL BE USED TO ASSESS EFFECT ON REACTIVATION VIA RNA-FISH AND QRT-PCR ASSAYS. OUR STUDIES WILL ESTABLISH INI1:TAR INTERACTION AS A DRUG TARGET. INHIBITING THIS INTERACTION COULD BLOCK LATENCY REACTIVATION PROMOTING DEEP LATENCY AND ADVANCING CURE STRATEGIES.

Albert Einstein College Of Medicine

<P>THE GOALS ARE:</P><UL><LI>TO FOSTER FUNDAMENTAL CREATIVE DISCOVERIES, INNOVATIVE RESEARCH STRATEGIES, AND THEIR APPLICATIONS AS A BASIS FOR ULTIMATELY PROTECTING AND IMPROVING HEALTH;</LI><LI>TO DEVELOP, MAINTAIN, AND RENEW SCIENTIFIC HUMAN AND PHYSICAL RESOURCES THAT WILL ENSURE THE NATION'S CAPABILITY TO PREVENT DISEASE;</LI><LI>TO EXPAND THE KNOWLEDGE BASE IN MEDICAL AND ASSOCIATED SCIENCES IN ORDER TO ENHANCE THE NATION'S ECONOMIC WELL-BEING AND ENSURE A CONTINUED HIGH RETURN ON THE PUBLIC INVESTMENT IN RESEARCH; AND</LI><LI>TO EXEMPLIFY AND PROMOTE THE HIGHEST LEVEL OF SCIENTIFIC INTEGRITY, PUBLIC ACCOUNTABILITY, AND SOCIAL RESPONSIBILITY IN THE CONDUCT OF SCIENCE.</LI></UL><P>IN REALIZING THESE GOALS, THE NIH PROVIDES LEADERSHIP AND DIRECTION TO PROGRAMS DESIGNED TO IMPROVE THE HEALTH OF THE NATION BY CONDUCTING AND SUPPORTING RESEARCH:</P><UL><LI>IN THE CAUSES, DIAGNOSIS, PREVENTION, AND CURE OF HUMAN DISEASES;</LI><LI>IN THE PROCESSES OF HUMAN GROWTH AND DEVELOPMENT;</LI><LI>IN THE BIOLOGICAL EFFECTS OF ENVIRONMENTAL CONTAMINANTS;</LI><LI>IN THE UNDERSTANDING OF MENTAL, ADDICTIVE AND PHYSICAL DISORDERS; AND</LI><LI>IN DIRECTING PROGRAMS FOR THE COLLECTION, DISSEMINATION, AND EXCHANGE OF INFORMATION IN MEDICINE AND HEALTH, INCLUDING THE DEVELOPMENT AND SUPPORT OF MEDICAL LIBRARIES AND THE TRAINING OF MEDICAL LIBRARIANS AND OTHER HEALTH INFORMATION SPECIALISTS.</LI></UL>

93.855 - Allergy and Infectious Diseases Research

National Institute of Allergy and Infectious Diseases (NIAID) [HHS - NIH]

Bronx, New York 10461 United States

Single Zip Code

NIH Research Project Grant (Parent R01 Clinical Trial Not Allowed) (PA-25-301)