Biochemical Characterization of Metnase's Endonuclease Activity and Its Role in NHEJ Repair

被引：38

作者：

Beck, Brian D. ^{[1
]}

Lee, Sung-Sook ^{[1
]}

Wiliamson, Elizabeth ^{[3
,4
]}

Hromas, Robert A. ^{[3
,4
]}

Lee, Suk-Hee ^{[1
,2
]}

机构：

[1] Indiana Univ, Sch Med, Dept Biochem & Mol Biol, Indianapolis, IN 46202 USA

[2] Indiana Univ, Sch Med, Simon Canc Ctr, Indianapolis, IN 46202 USA

[3] Univ Florida, Dept Med, Gainesville, FL 32610 USA

[4] Shands Hlth Care Syst, Gainesville, FL 32610 USA

来源：

BIOCHEMISTRY | 2011年 / 50卷 / 20期

关键词：

DOUBLE-STRAND BREAK; WERNER-SYNDROME PROTEIN; DNA-LIGASE IV; V(D)J RECOMBINATION; TRANSPOSASE DOMAIN; BINDING ACTIVITY; SETMAR PROTEIN; KINASE COMPLEX; KU COMPLEX; MECHANISM;

D O I：

10.1021/bi200333k

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Metnase (SETMAR) is a SET-transposase fusion protein that promotes nonhomologous end joining (NHEJ) repair in humans. Although both SET and the transposase domains were necessary for its function in DSB repair, it is not clear what specific role Metnase plays in the NHEJ. In this study, we show that Metnase possesses a unique endonuclease activity that preferentially acts on ssDNA and ssDNA-overhang of a partial duplex DNA. Cell extracts lacking Metnase poorly supported DNA end joining, and addition of wt-Metnase to cell extracts lacking Metnase markedly stimulated DNA end joining, while a mutant (D483A) lacking endonuclease activity did not. Given that Metnase overexpression enhanced DNA end processing in vitro, our finding suggests a role for Metnase's endonuclease activity in promoting the joining of noncompatible ends.

引用

页码：4360 / 4370

页数：11

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