In Vivo Roles of Rad52, Rad54, and Rad55 Proteins in Rad51-Mediated Recombination

• Published in: Molecular cell Vol. 12; no. 1; pp. 209 - 219
• Main Authors: Sugawara, Neal, Wang, Xuan, Haber, James E.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2003
• Subjects: Cells, Cultured; Chromosome Pairing - genetics; DNA - biosynthesis; DNA - genetics; DNA Helicases; DNA Repair - genetics; DNA Repair Enzymes; DNA, Single-Stranded - genetics; DNA, Single-Stranded - metabolism; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Mutation - genetics; Rad51 Recombinase; Rad52 DNA Repair and Recombination Protein; Recombination, Genetic - genetics; Repressor Proteins - genetics; Repressor Proteins - metabolism; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism
• ISSN: 1097-2765; 1097-4164
• DOI: 10.1016/S1097-2765(03)00269-7

Repairing a double-strand break by homologous recombination requires binding of the strand exchange protein Rad51p to ssDNA, followed by synapsis with a homologous donor. Here we used chromatin immunoprecipitation to monitor the in vivo association of Saccharomyces cerevisiae Rad51p with both the cleaved MATa locus and the HMLα donor. Localization of Rad51p to MAT precedes its association with HML, providing evidence of the time needed for the Rad51 filament to search the genome for a homologous sequence. Rad51p binding to ssDNA requires Rad52p. The absence of Rad55p delays Rad51p binding to ssDNA and prevents strand invasion and localization of Rad51p to HMLα. Lack of Rad54p does not significantly impair Rad51p recruitment to MAT or its initial association with HMLα; however, Rad54p is required at or before the initiation of DNA synthesis after synapsis has occurred at the 3′ end of the invading strand.