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The DNA binding preference of RAD52 and RAD59 proteins: implications for RAD52 and RAD59 protein function in homologous recombination.
J Biol Chem. 2006 Dec 29; 281(52):40001-9.JB

Abstract

We examined the double-stranded DNA (dsDNA) binding preference of the Saccharomyces cerevisiae Rad52 protein and its homologue, the Rad59 protein. In nuclease protection assays both proteins protected an internal sequence and the dsDNA ends equally well. Similarly, using electrophoretic mobility shift assays, we found the affinity of both Rad52 and Rad59 proteins for DNA ends to be comparable with their affinity for internal sequences. The protein-DNA complexes were also directly visualized using atomic force microscopy. Both proteins formed discrete complexes, which were primarily found (90-94%) at internal dsDNA sites. We also measured the DNA end binding behavior of human Rad52 protein and found a slight preference for dsDNA ends. Thus, these proteins have no strong preference for dsDNA ends over internal sites, which is inconsistent with their function at a step of dsDNA break repair that precedes DNA processing. Therefore, we conclude that S. cerevisiae Rad52 and Rad59 proteins and their eukaryotic counterparts function by binding to single-stranded DNA formed as intermediates of recombination rather than by binding to the unprocessed DNA double-strand break.

Authors+Show Affiliations

Section of Microbiology, Center for Genetics and Development, University of California, Davis, California 95616-8665, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

17040915

Citation

Wu, Yun, et al. "The DNA Binding Preference of RAD52 and RAD59 Proteins: Implications for RAD52 and RAD59 Protein Function in Homologous Recombination." The Journal of Biological Chemistry, vol. 281, no. 52, 2006, pp. 40001-9.
Wu Y, Siino JS, Sugiyama T, et al. The DNA binding preference of RAD52 and RAD59 proteins: implications for RAD52 and RAD59 protein function in homologous recombination. J Biol Chem. 2006;281(52):40001-9.
Wu, Y., Siino, J. S., Sugiyama, T., & Kowalczykowski, S. C. (2006). The DNA binding preference of RAD52 and RAD59 proteins: implications for RAD52 and RAD59 protein function in homologous recombination. The Journal of Biological Chemistry, 281(52), 40001-9.
Wu Y, et al. The DNA Binding Preference of RAD52 and RAD59 Proteins: Implications for RAD52 and RAD59 Protein Function in Homologous Recombination. J Biol Chem. 2006 Dec 29;281(52):40001-9. PubMed PMID: 17040915.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The DNA binding preference of RAD52 and RAD59 proteins: implications for RAD52 and RAD59 protein function in homologous recombination. AU - Wu,Yun, AU - Siino,Joseph S, AU - Sugiyama,Tomohiko, AU - Kowalczykowski,Stephen C, Y1 - 2006/10/12/ PY - 2006/10/17/pubmed PY - 2007/2/13/medline PY - 2006/10/17/entrez SP - 40001 EP - 9 JF - The Journal of biological chemistry JO - J Biol Chem VL - 281 IS - 52 N2 - We examined the double-stranded DNA (dsDNA) binding preference of the Saccharomyces cerevisiae Rad52 protein and its homologue, the Rad59 protein. In nuclease protection assays both proteins protected an internal sequence and the dsDNA ends equally well. Similarly, using electrophoretic mobility shift assays, we found the affinity of both Rad52 and Rad59 proteins for DNA ends to be comparable with their affinity for internal sequences. The protein-DNA complexes were also directly visualized using atomic force microscopy. Both proteins formed discrete complexes, which were primarily found (90-94%) at internal dsDNA sites. We also measured the DNA end binding behavior of human Rad52 protein and found a slight preference for dsDNA ends. Thus, these proteins have no strong preference for dsDNA ends over internal sites, which is inconsistent with their function at a step of dsDNA break repair that precedes DNA processing. Therefore, we conclude that S. cerevisiae Rad52 and Rad59 proteins and their eukaryotic counterparts function by binding to single-stranded DNA formed as intermediates of recombination rather than by binding to the unprocessed DNA double-strand break. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/17040915/The_DNA_binding_preference_of_RAD52_and_RAD59_proteins:_implications_for_RAD52_and_RAD59_protein_function_in_homologous_recombination_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(20)76834-1 DB - PRIME DP - Unbound Medicine ER -