title: A stable complex between integrase and viral DNA ends mediates human immunodeficiency virus integration in vitro.
abstract: Retroviral replication depends on integration of the viral genome into a chromosome of the host cell. The steps in this process are orchestrated in vivo by a large nucleoprotein complex and are catalyzed by the retroviral enzyme integrase. Several biochemical properties of the in vivo nucleoprotein complex were reproduced in vitro with purified integrase of human immunodeficiency virus type 1 and model viral DNA substrates. A stable complex between integrase and viral DNA was detected as an early intermediate in the integration reaction. After formation of this initial complex, the enzyme processively catalyzed the 3' end processing and strand transfer steps in the reaction. Complexes containing only purified integrase and the model viral DNA end were stable under a variety of conditions and efficiently used nonviral DNA molecules as integration targets. These complexes required a divalent cation for their formation, and their stability was highly dependent on the 5'-terminal dinucleotide of the viral DNA, for which no functional role has previously been defined. Thus, interactions between integrase and the extreme ends of the viral DNA molecule may be sufficient to account for the stability of the in vivo integration complex.
authors.affiliation: Department of Biochemistry, Stanford University, CA 94305-5428.
authors.forename: V
authors.initials: V
authors.lastname: Ellison
authors.forename: P O
authors.initials: PO
authors.lastname: Brown
mesh_terms: D001483:Base Sequence; D002413:Cations, Divalent; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D019426:Integrases; D008969:Molecular Sequence Data; D017384:Sequence Deletion; D016662:Virus Integration
chemical_list: D002413:Cations, Divalent; D004279:DNA, Viral; D004254:DNA Nucleotidyltransferases; D019426:Integrases