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PMID: 2574863 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Requirement for two DNA polymerases in the replication of simian virus 40 DNA in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 24 ·1989-12-00 ·Pages 9742-6

Weinberg DH, Kelly TJ

Abstract

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.

MeSH Terms
Antibodies, Monoclonal Chromatography, Ion Exchange DNA Polymerase III DNA Primase DNA Replication DNA-Directed DNA Polymerase/isolation & purification,metabolism Deoxyguanine Nucleotides/pharmacology HeLa Cells/enzymology Humans Kinetics Nuclear Proteins/isolation & purification Nucleic Acid Synthesis Inhibitors Proliferating Cell Nuclear Antigen RNA Nucleotidyltransferases/isolation & purification,metabolism Simian virus 40/genetics
Chemicals
Antibodies, Monoclonal Deoxyguanine Nucleotides Nuclear Proteins Nucleic Acid Synthesis Inhibitors Proliferating Cell Nuclear Antigen N(2)-(4-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate DNA Primase RNA Nucleotidyltransferases DNA Polymerase III DNA-Directed DNA Polymerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Weinberg D H
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Kelly T J
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31 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.24.9742
Published
1989-12-00
Pages
9742-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298577
Subset
IM
Grants
NCI NIH HHS · 5 RO1 CA40414 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.24.9742 PMC Full Text View journal details
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