Journal article
Analysis of the Human Replication Protein A:Rad52 Complex: Evidence for Crosstalk Between RPA32, RPA70, Rad52 and DNA
Journal of molecular biology, Vol.321(1), pp.133-148
2002
DOI: 10.1016/S0022-2836(02)00541-7
PMID: 12139939
Abstract
The eukaryotic single-stranded DNA-binding protein, replication protein A (RPA), is essential for DNA replication, and plays important roles in DNA repair and DNA recombination. Rad52 and RPA, along with other members of the Rad52 epistasis group of genes, repair double-stranded DNA breaks (DSBs). Two repair pathways involve RPA and Rad52, homologous recombination and single-strand annealing. Two binding sites for Rad52 have been identified on RPA. They include the previously identified C-terminal domain (CTD) of RPA32 (residues 224–271) and the newly identified domain containing residues 169–326 of RPA70. A region on Rad52, which includes residues 218–303, binds RPA70 as well as RPA32. The N-terminal region of RPA32 does not appear to play a role in the formation of the RPA:Rad52 complex. It appears that the RPA32CTD can substitute for RPA70 in binding Rad52. Sequence homology between RPA32 and RPA70 was used to identify a putative Rad52-binding site on RPA70 that is located near DNA-binding domains A and B. Rad52 binding to RPA increases ssDNA affinity significantly. Mutations in DBD-D on RPA32 show that this domain is primarily responsible for the ssDNA binding enhancement. RPA binding to Rad52 inhibits the higher-order self-association of Rad52 rings. Implications for these results for the “hand-off” mechanism between protein–protein partners, including Rad51, in homologous recombination and single-strand annealing are discussed.
Details
- Title: Subtitle
- Analysis of the Human Replication Protein A:Rad52 Complex: Evidence for Crosstalk Between RPA32, RPA70, Rad52 and DNA
- Creators
- Doba Jackson - Department of Chemistry, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606-3390, USAKajari Dhar - Department of Biochemistry, University of Iowa College of Medicine, 51 Newton Road, Iowa City, IA 52242-1109, USAJames K Wahl - Department of Biology, University of Toledo, Toledo, OH 43606-3390, USAMarc S Wold - Department of Biochemistry, University of Iowa College of Medicine, 51 Newton Road, Iowa City, IA 52242-1109, USAGloria E.O Borgstahl - Department of Chemistry, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606-3390, USA
- Resource Type
- Journal article
- Publication Details
- Journal of molecular biology, Vol.321(1), pp.133-148
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/S0022-2836(02)00541-7
- PMID
- 12139939
- ISSN
- 0022-2836
- eISSN
- 1089-8638
- Language
- English
- Date published
- 2002
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984025282302771
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