title: Allosteric inhibition of human immunodeficiency virus integrase: late block during viral replication and abnormal multimerization involving specific protein domains.
abstract: HIV-1 replication in the presence of antiviral agents results in evolution of drug-resistant variants, motivating the search for additional drug classes. Here we report studies of GSK1264, which was identified as a compound that disrupts the interaction between HIV-1 integrase (IN) and the cellular factor lens epithelium-derived growth factor (LEDGF)/p75. GSK1264 displayed potent antiviral activity and was found to bind at the site occupied by LEDGF/p75 on IN by x-ray crystallography. Assays of HIV replication in the presence of GSK1264 showed only modest inhibition of the early infection steps and little effect on integration targeting, which is guided by the LEDGF/p75-IN interaction. In contrast, inhibition of late replication steps was more potent. Particle production was normal, but particles showed reduced infectivity. GSK1264 promoted aggregation of IN and preformed LEDGF/p75-IN complexes, suggesting a mechanism of inhibition. LEDGF/p75 was not displaced from IN during aggregation, indicating trapping of LEDGF/p75 in aggregates. Aggregation assays with truncated IN variants revealed that a construct with catalytic and C-terminal domains of IN only formed an open polymer associated with efficient drug-induced aggregation. These data suggest that the allosteric inhibitors of IN are promising antiviral agents and provide new information on their mechanism of action.
authors.forename: Kushol
authors.initials: K
authors.lastname: Gupta
authors.forename: Troy
authors.initials: T
authors.lastname: Brady
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Dyer
authors.forename: Nirav
authors.initials: N
authors.lastname: Malani
authors.forename: Young
authors.initials: Y
authors.lastname: Hwang
authors.forename: Frances
authors.initials: F
authors.lastname: Male
authors.forename: Robert T
authors.initials: RT
authors.lastname: Nolte
authors.forename: Liping
authors.initials: L
authors.lastname: Wang
authors.forename: Emile
authors.initials: E
authors.lastname: Velthuisen
authors.forename: Jerry
authors.initials: J
authors.lastname: Jeffrey
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Van Duyne
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000494:Allosteric Regulation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D018360:Crystallography, X-Ray; D019427:HIV Integrase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D019428:HIV Integrase Inhibitors; C504649:PSIP1 protein, human; D014157:Transcription Factors; D019427:HIV Integrase