The yeast recombinational repair protein Rad59 interacts with Rad52 and stimulates single-strand annealing

Genetics. 2001 Oct;159(2):515-25. doi: 10.1093/genetics/159.2.515.

Abstract

The yeast RAD52 gene is essential for homology-dependent repair of DNA double-strand breaks. In vitro, Rad52 binds to single- and double-stranded DNA and promotes annealing of complementary single-stranded DNA. Genetic studies indicate that the Rad52 and Rad59 proteins act in the same recombination pathway either as a complex or through overlapping functions. Here we demonstrate physical interaction between Rad52 and Rad59 using the yeast two-hybrid system and co-immunoprecipitation from yeast extracts. Purified Rad59 efficiently anneals complementary oligonucleotides and is able to overcome the inhibition to annealing imposed by replication protein A (RPA). Although Rad59 has strand-annealing activity by itself in vitro, this activity is insufficient to promote strand annealing in vivo in the absence of Rad52. The rfa1-D288Y allele partially suppresses the in vivo strand-annealing defect of rad52 mutants, but this is independent of RAD59. These results suggest that in vivo Rad59 is unable to compete with RPA for single-stranded DNA and therefore is unable to promote single-strand annealing. Instead, Rad59 appears to augment the activity of Rad52 in strand annealing.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • DNA Primers
  • DNA Repair / genetics*
  • DNA, Fungal / genetics*
  • DNA, Single-Stranded / genetics*
  • DNA-Binding Proteins / metabolism*
  • Fungal Proteins / metabolism*
  • Mutation
  • Precipitin Tests
  • Protein Binding
  • Rad52 DNA Repair and Recombination Protein
  • Recombination, Genetic*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins*
  • Sequence Deletion

Substances

  • DNA Primers
  • DNA, Fungal
  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Fungal Proteins
  • RAD52 protein, S cerevisiae
  • RAD59 protein, S cerevisiae
  • Rad52 DNA Repair and Recombination Protein
  • Saccharomyces cerevisiae Proteins