Role for the Mammalian Swi5-Sfr1 Complex in DNA Strand Break Repair through Homologous Recombination

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Abstract
In fission yeast, the Swi5-Sfr1 complex plays an important role in homologous recombination (HR), a pathway crucial for the maintenance of genomic integrity. Here we identify and characterize mammalian Swi5 and Sfr1 homologues. Mouse Swi5 and Sfr1 are nuclear proteins that form a complex in vivo and in vitro. Swi5 interacts in vitro with Rad51, the DNA strand-exchange protein which functions during HR. By generating Swi5−/− and Sfr1−/− embryonic stem cell lines, we found that both proteins are mutually interdependent for their stability. Importantly, the Swi5-Sfr1 complex plays a role in HR when Rad51 function is perturbed in vivo by expression of a BRC peptide from BRCA2. Swi5−/− and Sfr1−/− cells are selectively sensitive to agents that cause DNA strand breaks, in particular ionizing radiation, camptothecin, and the Parp inhibitor olaparib. Consistent with a role in HR, sister chromatid exchange induced by Parp inhibition is attenuated in Swi5−/− and Sfr1−/− cells, and chromosome aberrations are increased. Thus, Swi5-Sfr1 is a newly identified complex required for genomic integrity in mammalian cells with a specific role in the repair of DNA strand breaks. Our genome constantly undergoes DNA damage as a result of agents in the environment, as well as from metabolic processes. One method of repairing DNA damage is homologous recombination (HR), in which genetic information from a duplicate sequence (the sister chromatid) is copied into the damaged site in DNA. In model organisms (the yeasts), a protein complex termed Swi5-Sfr1 functions in DNA damage repair by HR. In this study, we characterize mouse homologues of this complex. We find that mouse cells lacking this complex are sensitive to DNA damaging agents, in particular, those that cause breaks in DNA strands and that serve as cancer chemotherapeutics. These cells also have increased numbers of chromosome aberrations when exposed to DNA damaging agents. Moreover, HR is decreased in Swi5 and Sfr1 mutant cells under conditions where the cell is challenged. Together, these results demonstrate a requirement for the Swi5-Sfr1 protein complex in maintaining genomic integrity in mammalian cells.

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