Role of teh Rad52 Amino-terminal DNA Binding Activity in DNA Strand Capture in Homologous Recombination

Idina Shi, Swee Chuang Lim Hallwyl, Changhyun Seong, Uffe Hasbro Mortensen, Rodney Rothstein, Patrick Sung

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Saccharomyces cerevisiae Rad52 protein promotes homologous recombination by nucleating the Rad51 recombinase onto replication protein A-coated single-stranded DNA strands and also by directly annealing such strands. We show that the purified rad52-R70A mutant protein, with a compromised amino-terminal DNA binding domain, is capable of Rad51 delivery to DNA but is deficient in DNA annealing. Results from chromatin immunoprecipitation experiments find that rad52-R70A associates with DNA double-strand breaks and promotes recruitment of Rad51 as efficiently as wild-type Rad52. Analysis of gene conversion intermediates reveals that rad52-R70A cells can mediate DNA strand invasion but are unable to complete the recombination event. These results provide evidence that DNA binding by the evolutionarily conserved amino terminus of Rad52 is needed for the capture of the second DNA end during homologous recombination.
    Original languageEnglish
    JournalJournal of Biological Chemistry
    Volume284
    Issue number48
    Pages (from-to)33275-33284
    ISSN0021-9258
    DOIs
    Publication statusPublished - 2009

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