title: The Competitive Interplay between Allosteric HIV-1 Integrase Inhibitor BI/D and LEDGF/p75 during the Early Stage of HIV-1 Replication Adversely Affects Inhibitor Potency.
abstract: Allosteric HIV-1 integrase inhibitors (ALLINIs) have recently emerged as a promising class of antiretroviral agents and are currently in clinical trials. In infected cells, ALLINIs potently inhibit viral replication by impairing virus particle maturation but surprisingly exhibit a reduced EC50 for inhibiting HIV-1 integration in target cells. To better understand the reduced antiviral activity of ALLINIs during the early stage of HIV-1 replication, we investigated the competitive interplay between a potent representative ALLINI, BI/D, and LEDGF/p75 with HIV-1 integrase. While the principal binding sites of BI/D and LEDGF/p75 overlap at the integrase catalytic core domain dimer interface, we show that the inhibitor and the cellular cofactor induce markedly different multimerization patterns of full-length integrase. LEDGF/p75 stabilizes an integrase tetramer through the additional interactions with the integrase N-terminal domain, whereas BI/D induces protein-protein interactions in C-terminal segments that lead to aberrant, higher-order integrase multimerization. We demonstrate that LEDGF/p75 binds HIV-1 integrase with significantly higher affinity than BI/D and that the cellular protein is able to reverse the inhibitor induced aberrant, higher-order integrase multimerization in a dose-dependent manner in vitro. Consistent with these observations, alterations of the cellular levels of LEDGF/p75 markedly affected BI/D EC50 values during the early steps of HIV-1 replication. Furthermore, genome-wide sequencing of HIV-1 integration sites in infected cells demonstrate that LEDGF/p75-dependent integration site selection is adversely affected by BI/D treatment. Taken together, our studies elucidate structural and mechanistic details of the interplay between LEDGF/p75 and BI/D during the early stage of HIV-1 replication.
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Lei
authors.initials: L
authors.lastname: Feng
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
authors.forename: Venkatasubramanian
authors.initials: V
authors.lastname: Dharmarajan
authors.affiliation: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute and Department of Medicine, Harvard Medical School , Boston, Massachusetts 02215, United States.
authors.forename: Erik
authors.initials: E
authors.lastname: Serrao
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Ashley
authors.initials: A
authors.lastname: Hoyte
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Ross C
authors.initials: RC
authors.lastname: Larue
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Alison
authors.initials: A
authors.lastname: Slaughter
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Amit
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Plumb
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Kessl
authors.affiliation: Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: James R
authors.initials: JR
authors.lastname: Fuchs
authors.affiliation: Perelman School of Medicine, Department of Microbiology, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.affiliation: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute and Department of Medicine, Harvard Medical School , Boston, Massachusetts 02215, United States.
authors.forename: Alan N
authors.initials: AN
authors.lastname: Engelman
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Griffin
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Mamuka
authors.initials: M
authors.lastname: Kvaratskhelia
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D057809:HEK293 Cells; D015658:HIV Infections / Q000188:drug therapy* / Q000378:metabolism* / Q000821:virology; D019427:HIV Integrase / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D019428:HIV Integrase Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; C119380:lens epithelium-derived growth factor; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1