Visualization of local DNA unwinding by Mre11/Rad50/Nbs1 using single-molecule FRET

被引：51

作者：

Cannon, Brian ^{[1
]}

Kuhnlein, Jeffrey ^{[2
,3
]}

Yang, Soo-Hyun ^{[2
,3
]}

Cheng, Anita ^{[4
]}

Schindler, Detlev ^{[5
]}

Stark, Jeremy M. ^{[4
]}

Russell, Rick ^{[1
]}

Paull, Tanya T. ^{[2
,3
]}

机构：

[1] Univ Texas Austin, Dept Chem & Biochem, Austin, TX 78712 USA

[2] Univ Texas Austin, Inst Cellular & Mol Biol, Dept Mol Genet & Microbiol, Austin, TX 78712 USA

[3] Univ Texas Austin, Howard Hughes Med Inst, Austin, TX 78712 USA

[4] City Hope Natl Med Ctr, Beckman Res Inst, Dept Radiat Biol, Duarte, CA 91010 USA

[5] Univ Wurzburg, Dept Human Genet, Biozentrum, D-97074 Wurzburg, Germany

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2013年 / 110卷 / 47期

基金：

美国国家卫生研究院;

关键词：

DNA structure; DNA-protein interaction; DOUBLE-STRAND BREAK; ATM KINASE-ACTIVITY; SACCHAROMYCES-CEREVISIAE; END-RESECTION; REPAIR PROTEINS; MRE11; COMPLEX; RAD50; BINDING; NBS1; MRE11-RAD50-XRS2;

D O I：

10.1073/pnas.1309816110

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：

The Mre11/Rad50/Nbs1 (MRN) complex initiates and coordinates DNA repair and signaling events at double-strand breaks. The interaction between MRN and DNA ends is critical for the recruitment of DNA-processing enzymes, end tethering, and activation of the ATM protein kinase. Here we visualized MRN binding to duplex DNA molecules using single-molecule FRET, and found that MRN unwinds 15-20 base pairs at the end of the duplex, holding the branched structure open for minutes at a time in an ATP-dependent reaction. A Rad50 catalytic domain mutant that is specifically deficient in this ATP-dependent opening is impaired in DNA end resection in vitro and in resection-dependent repair of breaks in human cells, demonstrating the importance of MRN-generated single strands in the repair of DNA breaks.

引用

页码：18868 / 18873

页数：6

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