Nucleic Acids Research Advance Access originally published online on September 3, 2009
Nucleic Acids Research 2009 37(19):6429-6438; doi:10.1093/nar/gkp709
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Nucleic Acids Research, 2009, Vol. 37, No. 19 6429-6438
Published by Oxford University Press 2009.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Genome Integrity, Repair and Replication |
Rad10 exhibits lesion-dependent genetic requirements for recruitment to DNA double-strand breaks in Saccharomyces cerevisiae
1Department of Chemistry and Biochemistry, California State University, Northridge, CA 91330-8262, 2Department of Genetics & Development, Columbia University Medical Center, New York, NY 10032-2704, USA, 3Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, DK-2200 Copenhagen N, Denmark and 4Laboratory of Molecular Pathology, Department of Pathology, University of Texas, Southwestern Medical Center at Dallas, Dallas, TX 75390-9072, USA
*To whom correspondence should be addressed. Tel: +1 818 677 4503; Fax: +1 818 677 4378; Email: paula.fischhaber{at}csun.edu
Received December 12, 2008. Revised August 6, 2009. Accepted August 9, 2009.
In the yeast Saccharomyces cerevisiae, the Rad1–Rad10 protein complex participates in nucleotide excision repair (NER) and homologous recombination (HR). During HR, the Rad1–Rad10 endonuclease cleaves 3' branches of DNA and aberrant 3' DNA ends that are refractory to other 3' processing enzymes. Here we show that yeast strains expressing fluorescently labeled Rad10 protein (Rad10-YFP) form foci in response to double-strand breaks (DSBs) induced by a site-specific restriction enzyme, I-SceI or by ionizing radiation (IR). Additionally, for endonuclease-induced DSBs, Rad10-YFP localization to DSB sites depends on both RAD51 and RAD52, but not MRE11 while IR-induced breaks do not require RAD51. Finally, Rad10-YFP colocalizes with Rad51-CFP and with Rad52-CFP at DSB sites, indicating a temporal overlap of Rad52, Rad51 and Rad10 functions at DSBs. These observations are consistent with a putative role of Rad10 protein in excising overhanging DNA ends after homology searching and refine the potential role(s) of the Rad1–Rad10 complex in DSB repair in yeast.