Role of the Srs2–Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae

Shirin S. Jenkins, Steven Gore, Xiaoge Guo, Jie Liu, Christopher Ede, Xavier Veaute, Sue Jinks-Robertson, Stephen C Kowalczykowski, Wolf-Dietrich Heyer

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Saccharomyces cerevisiae Srs2, in addition to its well-documented antirecombination activity, has been proposed to play a role in promoting synthesis-dependent strand annealing (SDSA). Here we report the identification and characterization of an SRS2 mutant with a single amino acid substitution (srs2-F891A) that specifically affects the Srs2 pro-SDSA function. This residue is located within the Srs2–Rad51 interaction domain and embedded within a protein sequence resembling a BRC repeat motif. The srs2-F891A mutation leads to a complete loss of interaction with Rad51 as measured through yeast two-hybrid analysis and a partial loss of interaction as determined through protein pull-down assays with purified Srs2, Srs2-F891A, and Rad51 proteins. Even though previous work has shown that internal deletions of the Srs2–Rad51 interaction domain block Srs2 antirecombination activity in vitro, the Srs2-F891A mutant protein, despite its weakened interaction with Rad51, exhibits no measurable defect in antirecombination activity in vitro or in vivo. Surprisingly, srs2-F891A shows a robust shift from noncrossover to crossover repair products in a plasmid-based gap repair assay, but not in an ectopic physical recombination assay. Our findings suggest that the Srs2 C-terminal Rad51 interaction domain is more complex than previously thought, containing multiple interaction sites with unique effects on Srs2 activity.

Original languageEnglish (US)
Pages (from-to)1133-1145
Number of pages13
JournalGenetics
Volume212
Issue number4
DOIs
StatePublished - Jan 1 2019

Keywords

  • Crossover control
  • DNA repair
  • Genome stability
  • Helicase
  • Protein interaction
  • Recombination

ASJC Scopus subject areas

  • Genetics

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    Jenkins, S. S., Gore, S., Guo, X., Liu, J., Ede, C., Veaute, X., Jinks-Robertson, S., Kowalczykowski, S. C., & Heyer, W-D. (2019). Role of the Srs2–Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae. Genetics, 212(4), 1133-1145. https://doi.org/10.1534/genetics.119.302337