Protein dynamics during presynaptic-complex assembly on individual single-stranded DNA molecules

Bryan Gibb, Ling F. Ye, Youngho Kwon, Hengyao Niu, Patrick Sung, Eric C. Greene

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

Homologous recombination is a conserved pathway for repairing double-stranded breaks, which are processed to yield single-stranded DNA overhangs that serve as platforms for presynaptic-complex assembly. Here we use single-molecule imaging to reveal the interplay between Saccharomyces cerevisiae RPA, Rad52 and Rad51 during presynaptic-complex assembly. We show that Rad52 binds RPA-ssDNA and suppresses RPA turnover, highlighting an unanticipated regulatory influence on protein dynamics. Rad51 binding extends the ssDNA, and Rad52-RPA clusters remain interspersed along the presynaptic complex. These clusters promote additional binding of RPA and Rad52. Our work illustrates the spatial and temporal progression of the association of RPA and Rad52 with the presynaptic complex and reveals a new RPA-Rad52-Rad51-ssDNA intermediate, with implications for how the activities of Rad52 and RPA are coordinated with Rad51 during the later stages of recombination.

Original languageEnglish (US)
Pages (from-to)893-900
Number of pages8
JournalNature Structural and Molecular Biology
Volume21
Issue number10
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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