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PMID: 1346062 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex.

Molecular and cellular biology ·Vol. 12 ·No. 1 ·1992-01-00 ·Pages 155-63

Fien K, Stillman B

Abstract

A number of proteins have been isolated from human cells on the basis of their ability to support DNA replication in vitro of the simian virus 40 (SV40) origin of DNA replication. One such protein, replication factor C (RFC), functions with the proliferating cell nuclear antigen (PCNA), replication protein A (RPA), and DNA polymerase delta to synthesize the leading strand at a replication fork. To determine whether these proteins perform similar roles during replication of DNA from origins in cellular chromosomes, we have begun to characterize functionally homologous proteins from the yeast Saccharomyces cerevisiae. RFC from S. cerevisiae was purified by its ability to stimulate yeast DNA polymerase delta on a primed single-stranded DNA template in the presence of yeast PCNA and RPA. Like its human-cell counterpart, RFC from S. cerevisiae (scRFC) has an associated DNA-activated ATPase activity as well as a primer-template, structure-specific DNA binding activity. By analogy with the phage T4 and SV40 DNA replication in vitro systems, the yeast RFC, PCNA, RPA, and DNA polymerase delta activities function together as a leading-strand DNA replication complex. Now that RFC from S. cerevisiae has been purified, all seven cellular factors previously shown to be required for SV40 DNA replication in vitro have been identified in S. cerevisiae.

MeSH Terms
Adenosine Triphosphatases/metabolism Base Sequence DNA Polymerase III/metabolism DNA Replication DNA, Fungal/biosynthesis DNA-Binding Proteins/isolation & purification,metabolism Electrophoresis, Polyacrylamide Gel Fungal Proteins/isolation & purification,metabolism Molecular Sequence Data Nuclear Proteins/metabolism Proliferating Cell Nuclear Antigen Saccharomyces cerevisiae/genetics
Chemicals
DNA, Fungal DNA-Binding Proteins Fungal Proteins Nuclear Proteins Proliferating Cell Nuclear Antigen DNA Polymerase III Adenosine Triphosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fien K
Cold Spring Harbor Laboratory, New York 11724.
Stillman B
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54 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.1.155-163.1992
Published
1992-01-00
Pages
155-63
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364079
Subset
IM
Grants
NIAID NIH HHS · AI20460 · United States
External Links
PubMed source DOI: 10.1128/mcb.12.1.155-163.1992 PMC Full Text View journal details
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