title: Rad51 ATP binding but not hydrolysis is required to recruit Rad10 in synthesis-dependent strand annealing sites inS. cerevisiae.
abstract: Several modes of eukaryotic of DNA double strand break repair (DSBR) depend on synapsis of complementary DNA. The Rad51 ATPase, theS. cerevisiaehomolog ofE. coliRecA, plays a key role in this process by catalyzing homology searching and strand exchange between an invading DNA strand and a repair template (e.g. sister chromatid or homologous chromosome). Synthesis dependent strand annealing (SDSA), a mode of DSBR, requires Rad51. Another repair enzyme, the Rad1-Rad10 endonuclease, acts in the final stages of SDSA, hydrolyzing 3' overhanging single-stranded DNA. Here we showin vivoby fluorescence microscopy that the ATP binding function of yeast Rad51 is required to recruit Rad10 SDSA sites indicating that Rad51 pre-synaptic filament formation must occur prior to the recruitment of Rad1-Rad10. Our data also show that Rad51 ATPase activity, an important step in Rad51 filament disassembly, is not absolutely required in order to recruit Rad1-Rad10 to DSB sites.
authors.affiliation: Department of Chemistry and Biochemistry, California State University Northridge, Northridge, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Karlin
authors.affiliation: Department of Chemistry and Biochemistry, California State University Northridge, Northridge, USA.
authors.forename: Paula L
authors.initials: PL
authors.lastname: Fischhaber
keywords: Double Strand Break Repair; Rad1; Rad10; Rad51; Synthesis Dependent Strand Annealing; Yeast