title: Assembly and catalysis of concerted two-end integration events by Moloney murine leukemia virus integrase.
abstract: Retroviral integration results in the stable and coordinated insertion of the two termini of the linear viral DNA into the host genome. An in vitro concerted two-end integration reaction catalyzed by the Moloney murine leukemia virus (M-MuLV) integrase (IN) was used to investigate the binding and coordination of the two viral DNA ends. Comparison of the two-end integration and strand transfer assays indicates that zinc is required for efficient concerted integration utilizing plasmid DNA as target. Complementation assays using a pair of nonoverlapping integrase domains, consisting of the HHCC domain and the core/C-terminal region, yielded products containing the correct 4-base target site duplication. The efficiency of the coordinated two-end integration varied depending on the order of addition of the individual protein and DNA components in the complementation assay. Two-end integration was most efficient when the long terminal repeat (LTR) was premixed with either the target DNA or the HHCC domain. The preference for two-end integration through preincubation of the HHCC finger with the viral DNA supports the role of this domain in the recognition and/or positioning of the LTR.
authors.affiliation: Department of Biochemistry, University of Medicine and Dentistry of New Jersey-- Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.forename: M J
authors.initials: MJ
authors.lastname: Roth
mesh_terms: D001665:Binding Sites; D004279:DNA, Viral / Q000378:metabolism; D019426:Integrases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008979:Moloney murine leukemia virus / Q000201:enzymology*; D009841:Oligonucleotides / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D020079:Terminal Repeat Sequences; D016662:Virus Integration; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D004279:DNA, Viral; D009841:Oligonucleotides; D011994:Recombinant Proteins; D019426:Integrases; D015032:Zinc