The budding yeast Mei5-Sae3 complex interacts with Rad51 and preferentially binds a DNA fork structure

Amanda F. Say, Le Anna L. Ledford, Deepti Sharma, Akhilesh K. Singh, Wing Kit Leung, Hilarie A. Sehorn, Hideo Tsubouchi, Patrick Sung, Michael G. Sehorn

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Meiotic homologous recombination in Saccharomyces cerevisiae involves formation of nucleoprotein filaments of Rad51 and Dmc1 that mediate DNA strand exchange between homologous chromosomes. The Mei5-Sae3 protein complex functions as a recombination mediator to promote nucleation of the Dmc1 recombinase onto replication protein A-coated single-stranded DNA. Here, we have expressed and purified the Mei5 protein, Sae3 protein and the Mei5-Sae3 complex for biochemical studies. We show the Mei5-Sae3 complex preferentially binds a fork-like DNA substrate to 3′ overhanging DNA, single-stranded DNA or double-stranded DNA. We demonstrate that Mei5 confers DNA binding activity to the Mei5-Sae3 complex. We determined Mei5-Sae3 interacts with the Rad51 recombinase through the N-terminal domain of Mei5. Unlike Rad52, Mei5-Sae3 lacks recombination mediator activity for Rad51. Importantly, we find that the Mei5-Sae3 complex does not harbor single-strand DNA annealing activity. These properties of the Mei5-Sae3 complex distinguishes it from the Rad52 protein, which serves as the mediator of Rad51 and is involved in the single-strand DNA annealing pathway of homologous recombination.

Original languageEnglish (US)
Pages (from-to)586-594
Number of pages9
JournalDNA Repair
Volume10
Issue number6
DOIs
StatePublished - Jun 10 2011
Externally publishedYes

Keywords

  • Mei5-Sae3
  • Meiotic recombination
  • Rad51
  • Recombination mediator

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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