1. i.Regulation of HIV transcription and mechanisims of latency. ii.viral – host interactions. iii.Use of lentiviruses for targeting specific cells for gene therpy application. Dan Levy - Role of SETD6 in modulating HIV latency. Alon Freidman – Targeting of lentiviruses to specific cells. Lost in transcription – mechanisms that drive HIV latency Ongoing Projects Ongoing Collaborations

2. The Problem - latency is a block for HIV eradication Latency - a reversibly low-productive state of infection, where infected cells retain the capacity to fully re- emerge and produce de-novo viral particles New HIV infections AIDS- related deaths HAART People Total: 34.2 million HIV infected People Latent HIV reservoirs that are refractory to therapy

3. What are the molecular mechanisms that regulate HIV latency ? Ways to reactivate latent HIV and eliminate viral reservoirs with HAART ? Regulation of HIV transcriptional activation Role of Positive transcription elongation b -PTEFb HYPOTHESIS

4. NF-AT TAFs… Active State HIV Provirus gag pol tat rev env nef vif vpu vpr 5’LTR Latent State Condensed chromatin HDACs YY1/LSF CBF-1 AP4 NF-AT TAFs… CpG islands met H3K27 me3 SWI/SNF remodeling HIV provirus CCR5/CXCR4 CD4 met P-TEFb HATs HKMT/ SUV39H1 G9a NF-  B Ac 3’LTRAc met nuc1 Ac PRC1/2 Remodeling EZH2 NF-  B A B H3K9 me2 Latency corresponds with transcription activation and chromatin state Open chromatin

5. HIV Tat - a master switch of viral transcription CDK9 Cyclin T1 LTR RNA Polymerase II P P P P P-TEFb HIV Tat TAR

6. Recruitment modes of P-TEFb to the viral promoter

7. PKC activators (Bryostatin, Prostratin) HDAC inhibitors ( SAHA) Bromodomain and extra-terminal (BET) bromodomain inhibitors (JQ1, I- BET) Hypertrophic or stress signals (UV), TCR ligation (IL-2/CD3 Ab) HIV infection Tat Brd4 MePCE LARP7 CycT1 Cdk9 HEXIM1 5’5’ 7SK snRNA P P T270 S278 Ub CycT1 Cdk9 Active state - free active P- TEFb Resting stat - inactive P-TEFb P T186 Cdk9 CycT1 P P Ac K380;386;390;404 P13K/ Akt 3’3’ P-TEFb equilibrium in cells modulates HIV latency

8. What are the molecular mechanisms that regulate HIV latency ? Ways to reactivate latent HIV and eliminate viral reservoirs ? Role of P-TEFb in establishment of HIV latency Mechanisms that promote viral gene activation identification of host factors that modulate HIV latency Regulation of Transcriptional activation 1 2 3 Role of chromatin modulation (collaboration of D.Levy) Screen for small molecules that can reactivate latent HIV Modes of recruitment of P- TEFb to the viral promoter. 4