TY - JOUR
T1 - PCNA Is Required for Initiation of Recombination-Associated DNA Synthesis by DNA Polymerase δ
AU - Li, Xuan
AU - Stith, Carrie M.
AU - Burgers, Peter M.
AU - Heyer, Wolf Dietrich
N1 - Funding Information:
We thank Steve Kowalczykowski, Neil Hunter, and Amitabh Nimonkar as well as members of the Heyer laboratory (Shannon Ceballos, Kirk Ehmsen, Clare Fasching, Ryan Janke, Jie Liu, Damon Meyer, Erin Schwartz, Jessica Sneeden, William Wright, and Xiao-Ping Zhang) for discussion and comments on the manuscript. We thank Rita Alexeeva for help with DNA preparations, as well as Jie Liu, Kirk Ehmsen, and Xiao-Ping Zhang for help with protein preparations. This study was supported by National Institutes of Health (grant GM32431 to P.M.B. and grants CA92276 and GM58015 to W.D.H.).
PY - 2009/11/25
Y1 - 2009/11/25
N2 - 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.
AB - 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.
KW - DNA
UR - http://www.scopus.com/inward/record.url?scp=70449672959&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2009.09.036
DO - 10.1016/j.molcel.2009.09.036
M3 - Article
C2 - 19941829
AN - SCOPUS:70449672959
SN - 1097-2765
VL - 36
SP - 704
EP - 713
JO - Molecular cell
JF - Molecular cell
IS - 4
ER -