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Journal article
Human Rad52 binds and wraps single-stranded DNA and mediates annealing via two hRad52-ssDNA complexes
Nucleic acids research, v 38(9), pp 2917-2930
May 2010
PMID: 20081207
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Rad52 promotes the annealing of complementary strands of DNA bound by replication protein A (RPA) during discrete repair pathways. Here, we used a fluorescence resonance energy transfer (FRET) between two fluorescent dyes incorporated into DNA substrates to probe the mechanism by which human Rad52 (hRad52) interacts with and mediates annealing of ssDNA-hRPA complexes. Human Rad52 bound ssDNA or ssDNA-hRPA complex in two, concentration-dependent modes. At low hRad52 concentrations, ssDNA was wrapped around the circumference of the protein ring, while at higher protein concentrations, ssDNA was stretched between multiple hRad52 rings. Annealing by hRad52 occurred most efficiently when each complementary DNA strand or each ssDNA-hRPA complex was bound by hRad52 in a wrapped configuration, suggesting homology search and annealing occur via two hRad52-ssDNA complexes. In contrast to the wild type protein, hRad52(RQK/AAA) and hRad52(1-212) mutants with impaired ability to bind hRPA protein competed with hRPA for binding to ssDNA and failed to counteract hRPA-mediated duplex destabilization highlighting the importance of hRad52-hRPA interactions in promoting efficient DNA annealing.
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Details
- Title
- Human Rad52 binds and wraps single-stranded DNA and mediates annealing via two hRad52-ssDNA complexes
- Creators
- Jill M Grimme - Engineer Research and Development CenterMasayoshi Honda - University of Illinois Urbana-ChampaignRebecca Wright - University of Illinois Urbana-ChampaignYusuke Okuno - University of Illinois Urbana-ChampaignEli Rothenberg - Howard Hughes Medical InstituteAlexander V Mazin - Drexel UniversityTaekjip Ha - University of Illinois Urbana-ChampaignMaria Spies - Howard Hughes Medical Institute
- Publication Details
- Nucleic acids research, v 38(9), pp 2917-2930
- Publisher
- Oxford University Press
- Grant note
- GM065367 / NIGMS NIH HHS CA100839 / NCI NIH HHS R01 GM065367 / NIGMS NIH HHS MH084119 / NIMH NIH HHS Howard Hughes Medical Institute
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000277994600021
- Scopus ID
- 2-s2.0-77953280876
- Other Identifier
- 991019168481304721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology