ATP hydrolysis catalyzed by human replication factor C requires participation of multiple subunits

被引:41
作者
Cai, JS
Yao, NN
Gibbs, E
Finkelstein, J
Phillips, B
O'Donnell, M
Hurwitz, J
机构
[1] Mem Sloan Kettering Canc Ctr, William Randolph Hearst Lab Radiat Biol, Program Mol Biol, New York, NY 10021 USA
[2] Howard Hughes Med Inst, New York, NY 10021 USA
[3] Rockefeller Univ, New York, NY 10021 USA
关键词
D O I
10.1073/pnas.95.20.11607
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Human replication factor C (hRFC) is a five-subunit protein complex (p140, p40, p38, p37, and p36) that acts to catalytically load proliferating cell nuclear antigen onto DNA, where it recruits DNA polymerase delta or epsilon to the primer terminus at the expense of ATP, leading to processive DNA synthesis. We have previously shown that a subcomplex of hRFC consisting of three subunits (p40, p37, and p36) contained DNA-dependent ATPase activity. However, it is not clear which subunit(s) hydrolyzes ATP, as all five subunits include potential ATP binding sites. In this report,,ve introduced point mutations in the putative ATP-binding sequences of each hRFC subunit and examined the properties of the resulting mutant hRFC complex and the ATPase activity of the hRFC or the p40 p37 p36 complex. A mutation in any one of the ATP binding sites of the p36, p37, p40, or p140 subunits markedly reduced replication activity of the hRFC complex and the ATPase activity of the hRFC or the p40 p37 p36 complex. A mutation in the ATP binding site of the p38 subunit did not alter the replication activity of hRFC, These findings indicate that the replication activity of hRFC is dependent on efficient ATP hydrolysis contributed to by the action of four hRFC subunits.
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页码:11607 / 11612
页数:6
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