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PMID: 1320195 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Binding properties of replication protein A from human and yeast cells.

Molecular and cellular biology ·Vol. 12 ·No. 7 ·1992-07-00 ·Pages 3050-9

Kim C, Snyder RO, Wold MS

Abstract

Replication protein A (RP-A; also known as replication factor A and human SSB), is a single-stranded DNA-binding protein that is required for simian virus 40 DNA replication in vitro. RP-A isolated from both human and yeast cells is a very stable complex composed of 3 subunits (70, 32, and 14 kDa). We have analyzed the DNA-binding properties of both human and yeast RP-A in order to gain a better understanding of their role(s) in DNA replication. Human RP-A has high affinity for single-stranded DNA and low affinity for RNA and double-stranded DNA. The apparent affinity constant of RP-A for single-stranded DNA is in the range of 10(9) M-1. RP-A has a binding site size of approximately 30 nucleotides and does not bind cooperatively. The binding of RP-A to single-stranded DNA is partially sequence dependent. The affinity of human RP-A for pyrimidines is approximately 50-fold higher than its affinity for purines. The binding properties of yeast RP-A are similar to those of the human protein. Both yeast and human RP-A bind preferentially to the pyrimidine-rich strand of a homologous origin of replication: the ARS307 or the simian virus 40 origin of replication, respectively. This asymmetric binding suggests that RP-A could play a direct role in the process of initiation of DNA replication.

MeSH Terms
Base Sequence Binding, Competitive DNA Replication DNA-Binding Proteins/metabolism Humans Molecular Sequence Data Oligodeoxyribonucleotides/metabolism Polyribonucleotides/metabolism Replication Protein A Saccharomyces cerevisiae/metabolism Simian virus 40/metabolism
Chemicals
DNA-Binding Proteins Oligodeoxyribonucleotides Polyribonucleotides RPA1 protein, human Replication Protein A oligo (dT)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kim C
Department of Biochemistry, University of Iowa, Iowa City 52242.
Snyder R O
Wold M S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.7.3050-3059.1992
Published
1992-07-00
Pages
3050-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364519
Subset
IM
Grants
NIGMS NIH HHS · 1F32 GM14185-01 · United States
NIGMS NIH HHS · GM44721 · United States
External Links
PubMed source DOI: 10.1128/mcb.12.7.3050-3059.1992 PMC Full Text View journal details
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