Detection and characterization of a functional complex of human immunodeficiency virus type 1 integrase and its DNA substrate by UV cross-linking.
To analyze the association of the human immunodeficiency virus type 1 integrase (IN) protein with DNA substrates, we applied a shortwave UV cross-linking method to the in vitro integration system. Three photoadduct bands were detected on sodium dodecyl sulfate-polyacrylamide gels. The appearances of these photoadducts were examined with various substrates and under several incubation conditions. Our data suggest that one of these photoadducts is derived from the sequence- and strand-specific bound state of the early phase of the integration reaction before the 3' processing reaction. We also show that most of the photoadduct complexes are competent for integration even after UV irradiation.
Yoshinaga
T
T
Shionogi Institute for Medical Science, Osaka, Japan.
Kimura-Ohtani
Y
Y
Fujiwara
T
T
D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005033:Ethylmaleimide / Q000737:chemistry; D015497:HIV-1 / Q000201:enzymology*; D019426:Integrases; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010777:Photochemistry; D013329:Structure-Activity Relationship; D013997:Time Factors; D014466:Ultraviolet Rays; D016662:Virus Integration
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D005033:Ethylmaleimide