title: HIV-1 integrase complexes with DNA dissociate in the presence of short oligonucleotides conjugated to acridine.
abstract: The human immunodeficiency virus type 1 (HIV-1) integrase is an essential enzyme in the life cycle of the virus and is therefore an attractive target for the development of new antiviral drugs. Among them, inhibitors which are capable of targeting the preassembled integrase/DNA complex are of particular interest, because they could suppress integrase activity in the context of the HIV-1 preintegration complex. Here, we study the mechanism of action of 11-mer oligonucleotides, which are efficient inhibitors of the catalytic activity of integrase, provided that they are conjugated to a hydrophobic compound, acridine. To understand the mechanism of the conjugate inhibitory action, we used a steady-state fluorescence anisotropy assay, which allowed us to study the stability of the integrase/DNA complex in various conditions. We found that oligonucleotide-acridine conjugates induced the efficient dissociation of preassembled integrase/DNA complexes. The simultaneous presence of both acridine and an oligonucleotidic moiety is required for the inhibitory activity of conjugates. However, the dissociation effect is not dependent on the oligonucleotide sequence. Finally, our results suggest that the conjugates bind directly to integrase within its complex with DNA at a site different from the viral DNA binding site.
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119992 Moscow, Russia.
authors.forename: Marina
authors.initials: M
authors.lastname: Pinskaya
authors.forename: Elena
authors.initials: E
authors.lastname: Romanova
authors.forename: Evgenii
authors.initials: E
authors.lastname: Volkov
authors.forename: Eric
authors.initials: E
authors.lastname: Deprez
authors.forename: Hervé
authors.initials: H
authors.lastname: Leh
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Brochon
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouscadet
authors.forename: Marina
authors.initials: M
authors.lastname: Gottikh
mesh_terms: D000166:Acridines / Q000737:chemistry / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D019427:HIV Integrase / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects
chemical_list: D000166:Acridines; D019428:HIV Integrase Inhibitors; D009841:Oligonucleotides; D006493:Heparin; D004247:DNA; D019427:HIV Integrase