title: Complementation between HIV integrase proteins mutated in different domains.
abstract: HIV integrase (IN) cleaves two nucleotides off the 3' end of viral DNA and integrates viral DNA into target DNA. Previously, three functional domains in the HIV IN protein have been identified: (i) the central catalytic domain, (ii) the C-terminal DNA binding domain, and (iii) the N-terminal region, which is also necessary for activity. We have now investigated whether IN proteins mutated in different domains can complement each other. Mutant D116I does not contain an intact active site, but does bind DNA, whereas the C-terminal deletion mutant C delta 73 does not bind DNA, but does have an intact active site. Neither mutant protein mediates site-specific cleavage or integration. However, a mixture of both proteins is active, suggesting that IN functions as an oligomer, and that two subunits can have different functions; one subunit binds the (viral) DNA and another subunit provides the active site. We found three classes of mutants, corresponding to the three domains mentioned above. Mutants from different classes, but not from the same class, can complement each other. However, complementation is most efficient when the N- and C-termini are present on the same molecule.
authors.affiliation: Division of Molecular Biology, Netherlands Cancer Institute, Amsterdam.
authors.forename: D C
authors.initials: DC
authors.lastname: van Gent
authors.forename: C
authors.initials: C
authors.lastname: Vink
authors.forename: A A
authors.initials: AA
authors.lastname: Groeneger
authors.forename: R H
authors.initials: RH
authors.lastname: Plasterk
mesh_terms: D001665:Binding Sites / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D005816:Genetic Complementation Test; D015497:HIV-1 / Q000201:enzymology; D015498:HIV-2 / Q000201:enzymology; D019426:Integrases; D009154:Mutation; D012191:Retroviridae Proteins / Q000235:genetics*; D017384:Sequence Deletion
chemical_list: D004279:DNA, Viral; D012191:Retroviridae Proteins; D004254:DNA Nucleotidyltransferases; D019426:Integrases