title: Structural basis for HIV-1 DNA integration in the human genome, role of the LEDGF/P75 cofactor.
abstract: Integration of the human immunodeficiency virus (HIV-1) cDNA into the human genome is catalysed by integrase. Several studies have shown the importance of the interaction of cellular cofactors with integrase for viral integration and infectivity. In this study, we produced a stable and functional complex between the wild-type full-length integrase (IN) and the cellular cofactor LEDGF/p75 that shows enhanced in vitro integration activity compared with the integrase alone. Mass spectrometry analysis and the fitting of known atomic structures in cryo negatively stain electron microscopy (EM) maps revealed that the functional unit comprises two asymmetric integrase dimers and two LEDGF/p75 molecules. In the presence of DNA, EM revealed the DNA-binding sites and indicated that, in each asymmetric dimer, one integrase molecule performs the catalytic reaction, whereas the other one positions the viral DNA in the active site of the opposite dimer. The positions of the target and viral DNAs for the 3' processing and integration reaction shed light on the integration mechanism, a process with wide implications for the understanding of viral-induced pathologies.
authors.affiliation: IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Département de Biologie et de Génomique Structurales, UDS, CNRS, INSERM, Illkirch, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Michel
authors.forename: Corinne
authors.initials: C
authors.lastname: Crucifix
authors.forename: Florence
authors.initials: F
authors.lastname: Granger
authors.forename: Sylvia
authors.initials: S
authors.lastname: Eiler
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouscadet
authors.forename: Sergei
authors.initials: S
authors.lastname: Korolev
authors.forename: Julia
authors.initials: J
authors.lastname: Agapkina
authors.forename: Rustam
authors.initials: R
authors.lastname: Ziganshin
authors.forename: Marina
authors.initials: M
authors.lastname: Gottikh
authors.forename: Alexis
authors.initials: A
authors.lastname: Nazabal
authors.forename: Stéphane
authors.initials: S
authors.lastname: Emiliani
authors.forename: Richard
authors.initials: R
authors.lastname: Benarous
authors.forename: Dino
authors.initials: D
authors.lastname: Moras
authors.forename: Patrick
authors.initials: P
authors.lastname: Schultz
authors.forename: Marc
authors.initials: M
authors.lastname: Ruff
mesh_terms: D020285:Cryoelectron Microscopy; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015894:Genome, Human; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D008958:Models, Molecular; D011487:Protein Conformation; D016662:Virus Integration; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D036341:Intercellular Signaling Peptides and Proteins; C119380:lens epithelium-derived growth factor; D019427:HIV Integrase