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PMID: 8097277 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Primer-DNA formation during simian virus 40 DNA replication in vitro.

Molecular and cellular biology ·Vol. 13 ·No. 5 ·1993-05-00 ·Pages 2882-90

Denis D, Bullock PA

Abstract

Studies of simian virus 40 (SV40) DNA replication in vitro have identified a small (approximately 30-nucleotide) RNA-DNA hybrid species termed primer-DNA. Initial experiments indicated that T antigen and the polymerase alpha-primase complex are required to form primer-DNA. Proliferating cell nuclear antigen, and presumably proliferating cell nuclear antigen-dependent polymerases, is not needed to form this species. Herein, we present an investigation of the stages at which primer-DNA functions during SV40 DNA replication in vitro. Hybridization studies indicate that primer-DNA is initially formed in the origin region and is subsequently synthesized in regions distal to the origin. At all time points, primer-DNA is synthesized from templates for lagging-strand DNA replication. These studies indicate that primer-DNA functions during both initiation and elongation stages of SV40 DNA synthesis. Results of additional experiments suggesting a precursor-product relationship between formation of primer-DNA and Okazaki fragments are presented.

MeSH Terms
Antigens, Neoplasm/metabolism Antigens, Polyomavirus Transforming/metabolism Base Sequence Cell-Free System DNA Primase DNA Replication DNA, Viral/biosynthesis,isolation & purification HeLa Cells Humans Molecular Sequence Data Nuclear Proteins/metabolism Oligodeoxyribonucleotides Proliferating Cell Nuclear Antigen RNA Nucleotidyltransferases/metabolism Restriction Mapping Simian virus 40/genetics Templates, Genetic
Chemicals
Antigens, Neoplasm Antigens, Polyomavirus Transforming DNA, Viral Nuclear Proteins Oligodeoxyribonucleotides Proliferating Cell Nuclear Antigen DNA Primase RNA Nucleotidyltransferases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Denis D
Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111.
Bullock P A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.5.2882-2890.1993
Published
1993-05-00
Pages
2882-90
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359681
Subset
IM
Grants
NIAID NIH HHS · R01 AI30032 · United States
External Links
PubMed source DOI: 10.1128/mcb.13.5.2882-2890.1993 PMC Full Text View journal details
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