The yeast recombinational repair protein Rad59 interacts with Rad52 and stimulates single-strand annealing.
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.
Mesh Terms:
Base Sequence, DNA Primers, DNA Repair, DNA, Fungal, DNA, Single-Stranded, DNA-Binding Proteins, Fungal Proteins, Mutation, Precipitin Tests, Protein Binding, Rad52 DNA Repair and Recombination Protein, Recombination, Genetic, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Sequence Deletion
Base Sequence, DNA Primers, DNA Repair, DNA, Fungal, DNA, Single-Stranded, DNA-Binding Proteins, Fungal Proteins, Mutation, Precipitin Tests, Protein Binding, Rad52 DNA Repair and Recombination Protein, Recombination, Genetic, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Sequence Deletion
Genetics
Date: Oct. 01, 2001
PubMed ID: 11606529
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