Recruitment of the recombinational repair machinery to a DNA double-strand break in yeast

Branden Wolner, Stephen Van Komen, Patrick Sung, Craig L. Peterson

Research output: Contribution to journalArticle

159 Scopus citations

Abstract

Repair of DNA double-strand breaks (DSBs) by homologous recombination requires members of the RAD52 epistasis group. Here we use chromatin immunoprecipitation (ChIP) to examine the temporal order of recruitment of Rad51p, Rad52p, Rad54p, Rad55p, and RPA to a single, induced DSB in yeast. Our results suggest a sequential, interdependent assembly of Rad proteins adjacent to the DSB initiated by binding of Rad51p. ChIP time courses from various mutant strains and additional biochemical studies suggest that Rad52p, Rad55p, and Rad54p each help promote the formation and/or stabilization of the Rad51p nucleoprotein filament. We also find that all four Rad proteins associate with homologous donor sequences during strand invasion. These studies provide a near comprehensive view of the molecular events required for the in vivo assembly of a functional Rad51p presynaptic filament.

Original languageEnglish (US)
Pages (from-to)221-232
Number of pages12
JournalMolecular Cell
Volume12
Issue number1
DOIs
StatePublished - Jul 1 2003

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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