Interaction of AD2+D2 protein and simian virus 40 large T antigen with the large tumor antigen binding site I.
In a lytic infection of a permissive host by SV40, the large tumor antigen (T antigen), which is a product of early transcription of the SV40 A gene, has been previously shown to autoregulate its own transcription by binding to SV40 DNA. The DNA region to which T antigen binds most tightly was synthesized and subsequently introduced into the bacterial plasmid pUC8. The interaction of SV40 T antigen with the DNA duplexes, derived from both chemical synthesis and the recombinant plasmid, were examined by using nitrocellulose filter binding assays. An SV40-adenovirus hybrid protein, AD2+D2 protein, was also tested. The SV40 T antigen was found to bind more tightly than the hybrid protein. Kinetic assays demonstrated that the association rates for the two proteins with the DNA binding site were equivalent; however, once formed, the T antigen-DNA complex dissociated more slowly than the AD2+D2 protein-DNA complex.
D000957:Antigens, Viral, Tumor / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D004274:DNA, Recombinant / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D007700:Kinetics; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D010766:Phosphorylation; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology
D000957:Antigens, Viral, Tumor; D004274:DNA, Recombinant; D004279:DNA, Viral; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides