title: ATP stimulates the binding of simian virus 40 (SV40) large tumor antigen to the SV40 origin of replication.
abstract: Simian virus 40 (SV40) large tumor antigen (T antigen) binds to two contiguous sites at the SV40 origin of replication. Of these two sites, I and II, only site II is critical for replication. We have studied the interaction between T antigen and these sites by two methods--nitrocellulose filter binding and DNase I protection. We show that T antigen binds with high occupancy to site I at 0 degrees C, 25 degrees C, and 37 degrees C but to site II only at 0 degrees C and 25 degrees C. At 37 degrees C, the temperature essential for the initiation of SV40 DNA replication in vitro, ATP is required for the interaction of T antigen and site II. ATP can be replaced efficiently by adenosine 5'-[beta,gamma-imido]triphosphate and ADP, suggesting that hydrolysis of the nucleotide is not essential for the binding of T antigen to site II. The binding to the region critical for replication can occur in the presence of a variety of nucleoside triphosphates; dATP supports binding at a concentration 1/30th that of ATP, while dGTP and rGTP were inactive at all concentrations tested.
authors.affiliation: Graduate Program in Molecular Biology, Sloan-Kettering Cancer Center, New York, NY 10021.
authors.forename: J A
authors.initials: JA
authors.lastname: Borowiec
authors.forename: J
authors.initials: J
authors.lastname: Hurwitz
mesh_terms: D000227:Adenine Nucleotides / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D000952:Antigens, Polyomavirus Transforming; D002460:Cell Line; D004261:DNA Replication / Q000187:drug effects*; D004279:DNA, Viral / Q000187:drug effects / Q000235:genetics*; D003854:Deoxyribonucleotides / Q000494:pharmacology; D005814:Genes, Viral; D007700:Kinetics; D013539:Simian virus 40 / Q000187:drug effects / Q000235:genetics*; D013329:Structure-Activity Relationship
chemical_list: D000227:Adenine Nucleotides; D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D003854:Deoxyribonucleotides; D000255:Adenosine Triphosphate