title: In vitro functional analyses of the human immunodeficiency virus type 1 (HIV-1) integrase mutants give new insights into the intasome assembly.
abstract: A functional study of mutants of the human immunodeficiency virus type 1 (HIV-1) integrase (IN) was conducted with the support of a recently proposed HIV-1 intasome model. Firstly, we investigated the predicted position of the C-terminal domain (CTD) and the flexibility of the alpha-6 helix by mutating the residue Ile-203. This had no impact on the 3'-processing reaction but reduced the strand transfer reaction and the formation of tetramers. Secondly, the residues Ile-141 of the catalytic loop and Glu-246 of the CTD are predicted to bind the Td-3 base of the viral DNA maintaining it in a "flipped out" position and stabilizing the catalytic core domain (CCD)-CTD interface. Our data showed that the Ile-141/Td-3 interaction was important for the strand transfer activity and the oligomerization of IN. Interestingly, mutating the Glu-246 residue by an alanine enhanced half- and full-site integrations, suggesting that this residue may not be optimized for integration.
authors.affiliation: Laboratoire Rétrovirus et Pathologie Comparée, UMR 754, Institut National de la Recherche Agronomique, Université de Lyon, Université Lyon 1, Lyon, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Cellier
authors.forename: Karen
authors.initials: K
authors.lastname: Moreau
authors.forename: Kathy
authors.initials: K
authors.lastname: Gallay
authors.forename: Allison
authors.initials: A
authors.lastname: Ballandras
authors.forename: Patrice
authors.initials: P
authors.lastname: Gouet
authors.forename: Corinne
authors.initials: C
authors.lastname: Ronfort
mesh_terms: D019943:Amino Acid Substitution; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000378:metabolism; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D020125:Mutation, Missense; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1