Homotypic and heterotypic protein associations control Rad51 function in double-strand break repair.
Rad51 is essential for efficient repair of DNA double-strand breaks (DSBs) and recombination in Saccharomyces cerevisiae. Here, we examine Rad51 protein-protein interactions and their biological significance. GAL4 two-hybrid fusion analysis demonstrated that the amino-terminal region of Rad51 mediates both a strong Rad51:Rad51 self-association and a Rad51:Rad52 interaction. Several Rad51 variants ... were characterized that imparted DSB repair defects; these defects appear to result from Rad51 protein-protein interactions. First, a rad51 allele bearing a missense mutation in the consensus ATP-binding sequence disrupted DSB repair in wild-type yeast. The effect of this allele was dependent on the presence of wild-type Rad51 because MMS sensitivity of rad51 delta strains were not increased by its expression. Second, we identified a highly conserved RAD51 homolog from Kluyveromyces lactis (KlRAD51) that only partially complemented rad51 delta strains and impaired DSB repair in wild-type S. cerevisiae. Third, fusions of Gal4 domains to Rad51 disrupted DSB repair in a manner that required the presence of either Rad51 or Rad52. Because K. lactis RAD51 and RAD52 did not complement a S. cerevisiae rad51 delta rad52 delta strain, Rad51-Rad52 functions appear to be mediated through additional components. Thus, multiple types of Rad51 protein interactions, including self-association, appear to be important for DSB repair.
Mesh Terms:
Alleles, Amino Acid Sequence, Base Sequence, Cloning, Molecular, DNA Primers, DNA Repair, DNA-Binding Proteins, Fungal Proteins, Genes, Fungal, Kinetics, Kluyveromyces, Molecular Sequence Data, Phenotype, Polymerase Chain Reaction, Protein Multimerization, Rad51 Recombinase, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Transcription Factors, Two-Hybrid System Techniques
Alleles, Amino Acid Sequence, Base Sequence, Cloning, Molecular, DNA Primers, DNA Repair, DNA-Binding Proteins, Fungal Proteins, Genes, Fungal, Kinetics, Kluyveromyces, Molecular Sequence Data, Phenotype, Polymerase Chain Reaction, Protein Multimerization, Rad51 Recombinase, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Transcription Factors, Two-Hybrid System Techniques
Genes Dev.
Date: Nov. 01, 1994
PubMed ID: 7958917
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