Journal article
Binding properties of replication protein A from human and yeast cells
Molecular and cellular biology, Vol.12(7), pp.3050-3059
07/1992
DOI: 10.1128/mcb.12.7.3050-3059.1992
PMCID: PMC364519
PMID: 1320195
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.
Details
- Title: Subtitle
- Binding properties of replication protein A from human and yeast cells
- Creators
- Changsoo Kim - Department of Biochemistry, University of Iowa, Iowa City 52242R O Snyder - Department of Biochemistry, University of Iowa, Iowa City 52242Marc S. Wold - Department of Biochemistry, University of Iowa, Iowa City 52242
- Resource Type
- Journal article
- Publication Details
- Molecular and cellular biology, Vol.12(7), pp.3050-3059
- DOI
- 10.1128/mcb.12.7.3050-3059.1992
- PMID
- 1320195
- PMCID
- PMC364519
- ISSN
- 0270-7306
- eISSN
- 1098-5549
- Language
- English
- Date published
- 07/1992
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984025394902771
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