Requirement for two DNA polymerases in the replication of simian virus 40 DNA in vitro.
DNA polymerase alpha-primase has long been considered the primary, if not sole, replicative DNA polymerase in eukaryotic cells. However, recent experiments have provided indirect evidence that a second DNA polymerase may play a role in DNA replication. To identify cellular proteins necessary for DNA synthesis in mammalian cells, we have been studying the cell-free system developed for the replication of simian virus 40 DNA. In this report, we present direct evidence that a second DNA polymerase is required in addition to DNA polymerase alpha-primase complex to obtain efficient replication of simian virus 40 origin-containing DNA. This DNA polymerase activity is not affected by monoclonal antibodies that inhibit the activity of DNA polymerase alpha and is relatively resistant to the inhibitor [N2-(p-n-butylphenyl)-9-(2-deoxy-beta-D-ribofuranosyl)guanine 5'-triphosphate]. Moreover, the activity of the polymerase is highly dependent upon the accessory protein, proliferating-cell nuclear antigen. These characteristics are consistent with the hypothesis that this second DNA polymerase is DNA polymerase delta.
Weinberg
D H
DH
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Kelly
T J
TJ
D000911:Antibodies, Monoclonal; D002852:Chromatography, Ion Exchange; D004258:DNA Polymerase III; D019915:DNA Primase; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D009687:Nuclear Proteins / Q000302:isolation & purification; D019384:Nucleic Acid Synthesis Inhibitors; D018809:Proliferating Cell Nuclear Antigen; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D013539:Simian virus 40 / Q000235:genetics*
D000911:Antibodies, Monoclonal; D003848:Deoxyguanine Nucleotides; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D018809:Proliferating Cell Nuclear Antigen; C040364:N(2)-(4-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase