ATP stimulates the binding of simian virus 40 (SV40) large tumor antigen to the SV40 origin of replication.
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.
Borowiec
J A
JA
Graduate Program in Molecular Biology, Sloan-Kettering Cancer Center, New York, NY 10021.
Hurwitz
J
J
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
D000227:Adenine Nucleotides; D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D003854:Deoxyribonucleotides; D000255:Adenosine Triphosphate