Functional interactions among yeast Rad51 recombinase, Rad52 mediator, and replication protein A in DNA strand exchange.
Rad51-catalyzed DNA strand exchange is greatly enhanced by the single-stranded (ss) DNA binding factor RPA if the latter is introduced after Rad51 has already nucleated onto the initiating ssDNA substrate. Paradoxically, co-addition of RPA with Rad51 to the ssDNA to mimic the in vivo situation diminishes the level of strand exchange, revealing competition between RPA and Rad51 for binding sites on ssDNA. Rad52 promotes strand exchange but only when there is a need for Rad51 to compete with RPA for loading onto ssDNA. Rad52 is multimeric, binds ssDNA, and targets Rad51 to ssDNA. Maximal restoration of pairing and strand exchange requires amounts of Rad52 substoichiometric to Rad51 and involves a stable, equimolar complex between Rad51 and Rad52. The Rad51-Rad52 complex efficiently utilizes a ssDNA template saturated with RPA for homologous pairing but does not appear to be more active than Rad51 when an RPA-free ssDNA template is used. Rad52 does not substitute for RPA in the pairing and strand exchange reaction nor does it lower the dependence of the reaction on Rad51 or RPA.
Song
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B
Department of Molecular Medicine/Institute of Biotechnology, University of Texas Health Science Center, San Antonio, Texas 78245-3207, USA.
Sung
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D000251:Adenosine Triphosphatases / Q000378:metabolism; D010584:Bacteriophage phi X 174 / Q000235:genetics; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism; D011485:Protein Binding; D051135:Rad51 Recombinase; D051721:Rad52 DNA Repair and Recombination Protein; D011995:Recombination, Genetic / Q000235:genetics*; D051716:Replication Protein A; D015003:Yeasts
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D051721:Rad52 DNA Repair and Recombination Protein; D051716:Replication Protein A; D004247:DNA; D051135:Rad51 Recombinase; D000251:Adenosine Triphosphatases