DNA Replication-Coupled PCNA Mono-Ubiquitination and Polymerase Switching in a Human In Vitro System

被引：35
|
作者：
Masuda, Yuji [1 ]
Piao, Jinlian [1 ]
Kamiya, Kenji [1 ]
机构：
[1] Hiroshima Univ, Res Inst Radiat Biol & Med, Dept Expt Oncol, Minami Ku, Hiroshima 7348553, Japan
来源：
JOURNAL OF MOLECULAR BIOLOGY | 2010年 / 396卷 / 03期
基金：
日本学术振兴会;
关键词：
DNA polymerase switching; DNA replication; PCNA mono-ubiquitination; translesion DNA synthesis; CELL NUCLEAR ANTIGEN; TRANSLESION SYNTHESIS; BINDING DOMAINS; SCHIZOSACCHAROMYCES-POMBE; SACCHAROMYCES-CEREVISIAE; MONOUBIQUITINATED PCNA; GENOMIC INSTABILITY; COMPLEX-FORMATION; REPAIR PROTEINS; SLIDING CLAMP;
D O I：
10.1016/j.jmb.2010.01.003
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：
Translesion DNA synthesis is a mechanism of DNA damage tolerance, and mono-ubiquitination of proliferating cell nuclear antigen (PCNA) is considered to play a key role in regulating the switch from replicative to translesion DNA polymerases (pols). In this study, we analyzed effects of a replicative pol delta on PCNA mono-ubiquitination with the ubiquitin-conjugating enzyme and ligase UBE2A/HHR6A/RAD6A-RAD18. The results revealed that PCNA interacting with pol 8 is a better target for ubiquitination, and PCNA mono-ubiquitination could be coupled with DNA replication. Consequently, we could reconstitute replication-coupled switching between pol delta and a translesion pol, pol eta, on an ultraviolet-light-irradiated template. With this system, we obtained direct evidence that polymerase switching reactions are stimulated by mono-ubiquitination of PCNA, depending on a function of the ubiquitin binding zinc finger domain of pol eta. This study provides a framework for detailed analyses of molecular mechanisms of human pol switching and regulation of translesion DNA synthesis. (C) 2010 Elsevier Ltd. All rights reserved.
引用
收藏
页码：487 / 500
页数：14
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