title: Disulfide-linked integrase oligomers involving C280 residues are formed in vitro and in vivo but are not essential for human immunodeficiency virus replication.
abstract: The human immunodeficiency virus type 1 integrase (IN) forms an oligomer that integrates both ends of the viral DNA. The nature of the active oligomer is unclear. Recombinant IN obtained under reducing conditions is always in the form of noncovalent oligomers. However, disulfide-linked oligomers of IN were recently observed within viral particles. We show that IN produced from a baculovirus expression system can form disulfide-linked oligomers. We investigated which residues are responsible for the disulfide bridges and the relationship between the ability to form covalent dimers and IN activity. Only the mutation of residue C280 was sufficient to prevent the formation of intermolecular disulfide bridges in oligomers of recombinant IN. IN activity was studied under and versus nonreducing conditions: the formation of disulfide bridges was not required for the in vitro activities of the enzyme. Moreover, the covalent dimer does not dissociate into individual protomers on disulfide bridge reduction. Instead, IN undergoes a spontaneous multimerization process that yields a homogenous noncovalent tetramer. The C280S mutation also completely abolished the formation of disulfide bonds in the context of the viral particle. Finally, the replication of the mutant virus was investigated in replicating and arrested cells. The infectivity of the virus was not affected by the C280S IN mutation in either dividing or nondividing cells. The disulfide-linked form of the IN oligomers observed in the viral particles is thus not required for viral replication.
authors.affiliation: CNRS UMR8532, LBPA, Ecole Normale Supérieure de Cachan, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Bischerour
authors.forename: Hervé
authors.initials: H
authors.lastname: Leh
authors.forename: Eric
authors.initials: E
authors.lastname: Deprez
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Brochon
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouscadet
mesh_terms: D003545:Cysteine; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry*; D019427:HIV Integrase / Q000737:chemistry* / Q000502:physiology; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D014771:Virion / Q000737:chemistry; D014779:Virus Replication
chemical_list: D004220:Disulfides; D019427:HIV Integrase; D003545:Cysteine