西亚试剂：PCNA Is Required for Initiation of Recombination-Associated

PCNA Is Required for Initiation of Recombination-Associated DNA Synthesis by DNA Polymerase δ

Xuan Li1, 4, Carrie M. Stith3, Peter M. Burgers3 and Wolf-Dietrich Heyer1, 2, , 

1 Department of Microbiology, University of California, Davis, Davis, CA 95616-8665, USA
2 Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA 95616-8665, USA
3 Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110, USA

Genetic recombination ensures proper chromosome segregation during meiosis and is essential for genome stability and tumor suppression. DNA synthesis after Rad51-mediated DNA strand invasion is a crucial step during recombination. PCNA is known as the processivity clamp for DNA polymerases. Here, we report the surprising observation that PCNA is specifically required to initiate recombination-associated DNA synthesis in the extension of the 3′ end of the invading strand in a D loop. We show using a reconstituted system of yeast Rad51, Rad54, RPA, PCNA, RFC, and DNA polymerase δ that loading of PCNA by RFC targets DNA polymerase δ to the D loop formed by Rad51 protein, allowing efficient utilization of the invading 3′ end and processive DNA synthesis. We conclude that PCNA has a specific role in the initiation of recombination-associated DNA synthesis and that DNA polymerase δ promotes recombination-associated DNA synthesis.

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