Conserved residues Pro-109 and Asp-116 are required for interaction of the human immunodeficiency virus type 1 integrase protein with its viral DNA substrate.
The human immunodeficiency virus type 1 integrase protein can be specifically cross-linked to viral long terminal repeat substrate oligonucleotides in vitro by using UV light. Site-directed mutagenesis and deletion analyses were used to define the domains involved in the interaction of integrase with the viral DNA substrate. Our results showed that mutation of conserved residues Pro-109 and Asp-116, which are found to be critical for the endonuclease and integration activities of IN protein, abolished the ability of the protein to cross-link to its DNA substrate. Furthermore, deletion analysis experiments showed that removal of 39 amino acids from the amino terminus and deletion of 15 amino acids from the carboxyl terminus abolished DNA cross-linking.
Pharmaceutical Research New Technologies, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
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Drelich
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Haenggi
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Mous
D000595:Amino Acid Sequence; D001224:Aspartic Acid; D001483:Base Sequence; D017124:Conserved Sequence; D003432:Cross-Linking Reagents; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019426:Integrases; D009052:Leukemia Virus, Murine / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D011392:Proline; D012091:Repetitive Sequences, Nucleic Acid; D017384:Sequence Deletion; D013379:Substrate Specificity; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D001224:Aspartic Acid; D011392:Proline; D004254:DNA Nucleotidyltransferases; D019426:Integrases