The overall goal of our studies is to better understand structure and function of HIV-1 integrase (IN) as a therapeutic target. A multimer of IN stably associates with two viral DNA ends in the context of the preintegration complex (PIC) and catalyzes concerted integration of the viral DNA ends into the host chromosome, a process which is essential for retroviral replication. Lens Epithelium Derived Growth Factor (LEDGF/p75) is a key cellular binding partner of HIV-1 IN and contributes to effective integration by tethering PICs to active genes. During the past funding cycle the efforts in the PI's group focused on examining molecular interactions of IN with viral DNA substrates, LEDGF/p75 and small molecule inhibitors. We have mapped HIV-1 IN interactions with viral DNA and identified novel protein contacts essential for the 3'-processing reactions. Furthermore, our group has uncovered new inter- and intra-protein-protein contacts for the full length IN-LEDGF/p75 complex and demonstrated that the cellular cofactor strongly modulates highly dynamic structure and function of the retroviral enzyme. Moreover, our group identified a small molecule inhibitor of HIV-1 IN that selectively binds to the IN dimer interface. Collectively, these findings have laid a strong foundation for extending our efforts further to address the following new and important questions.
Aim 1 will identify functional contacts between IN, viral DNA and LEDGF/p75 in concerted integration intermediates using our MS-based footprinting approach.
Aim 2 will examine modulation of IN structure and function by LEDGF/p75 using protein-protein FRET approach.
Aim 3 will probe potential direct interactions between LEDGF PWWP and the modified histones using a histone peptide array.
Aim 4 will further investigate the mechanism of action for the small molecule allosteric inhibitor of IN by analyzing various derivatives of the lead compound. Taken together, these new findings will enhance our understanding how LEDGF/p75 modulates IN interactions with viral DNA and navigates the IN-viral DNA complex to the active transcription units at the chromatin. Furthermore, our studies with the allosteric inhibitor will contribute to wider efforts in the field focused on discovery of new antiretroviral therapies.

Public Health Relevance

HIV-1 integrase (IN) is commonly viewed as an important therapeutic target for the following reasons: its catalytic activities are required for the virus life cycle, there is no closely related human equivalent of IN, and specific IN inhibitors are likely to be effective against viral strains resistant to currently available therapies targeting HIV-1 reverse transcriptase, protease and fusion. The proposed studies will investigate molecular interactions of IN with viral DNA substrates, the key cellular cofactor LEDGF/p75 and small molecule inhibitors. The obtained results will improve our understanding of how this key retroviral enzyme functions and contribute to wider efforts in the field focused on discovery of new antiretroviral therapies. 1

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
AIDS Molecular and Cellular Biology Study Section (AMCB)
Program Officer
Sharma, Opendra K
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Ohio State University
Other Health Professions
Schools of Pharmacy
United States
Zip Code
Kessl, Jacques J; Kutluay, Sebla B; Townsend, Dana et al. (2016) HIV-1 Integrase Binds the Viral RNA Genome and Is Essential during Virion Morphogenesis. Cell 166:1257-1268.e12
Kessl, Jacques J; Sharma, Amit; Kvaratskhelia, Mamuka (2016) Methods for the Analyses of Inhibitor-Induced Aberrant Multimerization of HIV-1 Integrase. Methods Mol Biol 1354:149-64
Ngure, Marianne; Issur, Moheshwarnath; Shkriabai, Nikoloz et al. (2016) Interactions of the Disordered Domain II of Hepatitis C Virus NS5A with Cyclophilin A, NS5B, and Viral RNA Show Extensive Overlap. ACS Infect Dis 2:839-851
Crowe, Brandon L; Larue, Ross C; Yuan, Chunhua et al. (2016) Structure of the Brd4 ET domain bound to a C-terminal motif from γ-retroviral integrases reveals a conserved mechanism of interaction. Proc Natl Acad Sci U S A 113:2086-91
Chen, Tien-Hao; Tanimoto, Akiko; Shkriabai, Nikoloz et al. (2016) Use of chemical modification and mass spectrometry to identify substrate-contacting sites in proteinaceous RNase P, a tRNA processing enzyme. Nucleic Acids Res 44:5344-55
Feng, Lei; Larue, Ross C; Slaughter, Alison et al. (2015) HIV-1 integrase multimerization as a therapeutic target. Curr Top Microbiol Immunol 389:93-119
Singh, Parmit Kumar; Plumb, Matthew R; Ferris, Andrea L et al. (2015) LEDGF/p75 interacts with mRNA splicing factors and targets HIV-1 integration to highly spliced genes. Genes Dev 29:2287-97
Hilton, Benjamin; Shkriabai, Nick; Musich, Phillip R et al. (2014) A new structural insight into XPA-DNA interactions. Biosci Rep 34:e00162
Slaughter, Alison; Jurado, Kellie A; Deng, Nanjie et al. (2014) The mechanism of H171T resistance reveals the importance of Nδ-protonated His171 for the binding of allosteric inhibitor BI-D to HIV-1 integrase. Retrovirology 11:100
Kvaratskhelia, Mamuka; Sharma, Amit; Larue, Ross C et al. (2014) Molecular mechanisms of retroviral integration site selection. Nucleic Acids Res 42:10209-25

Showing the most recent 10 out of 32 publications