title: Rad10 exhibits lesion-dependent genetic requirements for recruitment to DNA double-strand breaks in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Department of Chemistry and Biochemistry, California State University, Northridge, CA 91330-8262, USA.
authors.forename: Destaye M
authors.initials: DM
authors.lastname: Moore
authors.forename: Justin
authors.initials: J
authors.lastname: Karlin
authors.forename: Sergio
authors.initials: S
authors.lastname: González-Barrera
authors.forename: Armen
authors.initials: A
authors.lastname: Mardiros
authors.forename: Michael
authors.initials: M
authors.lastname: Lisby
authors.forename: Ana
authors.initials: A
authors.lastname: Doughty
authors.forename: Jennifer
authors.initials: J
authors.lastname: Gilley
authors.forename: Rodney
authors.initials: R
authors.lastname: Rothstein
authors.forename: Errol C
authors.initials: EC
authors.lastname: Friedberg
authors.forename: Paula L
authors.initials: PL
authors.lastname: Fischhaber
mesh_terms: D053903:DNA Breaks, Double-Stranded; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C064162:RAD10 protein, S cerevisiae; D015719:Single-Strand Specific DNA and RNA Endonucleases