Synthesis of DNA containing the simian virus 40 origin of replication by the combined action of DNA polymerases alpha and delta.
Proliferating-cell nuclear antigen (PCNA) mediates the replication of simian virus 40 (SV40) DNA by reversing the effects of a protein that inhibits the elongation reaction. Two other protein fractions, activator I and activator II, were also shown to play important roles in this process. We report that activator II isolated from HeLa cell extracts is a PCNA-dependent DNA polymerase delta that is required for efficient replication of DNA containing the SV40 origin of replication. PCNA-dependent DNA polymerase delta on a DNA singly primed phi X174 single-stranded circular DNA template required PCNA, a complex of the elongation inhibitor and activator I, and the single-stranded DNA-binding protein essential for SV40 DNA replication. DNA polymerase delta, in contrast to DNA polymerase alpha, hardly used RNA-primed DNA templates. These results indicate that both DNA polymerase alpha and delta are involved in SV40 DNA replication in vitro and their activity depends on PCNA, the elongation inhibitor, and activator I.
Lee
S H
SH
Graduate Program in Molecular Biology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Eki
T
T
Hurwitz
J
J
D000951:Antigens, Neoplasm; D003600:Cytosol / Q000378:metabolism; D004257:DNA Polymerase II / Q000378:metabolism*; D004258:DNA Polymerase III; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009687:Nuclear Proteins / Q000502:physiology; D018809:Proliferating Cell Nuclear Antigen; D013539:Simian virus 40 / Q000235:genetics*; D013698:Templates, Genetic
D000951:Antigens, Neoplasm; D004279:DNA, Viral; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D004257:DNA Polymerase II; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase