Stalled replication forks generate a distinct mutational signature in yeast

被引:21
作者
Larsen, Nicolai B. [1 ,2 ,3 ]
Liberti, Sascha E. [1 ,2 ]
Vogel, Ivan [1 ]
Jorgensen, Signe W. [1 ,2 ]
Hickson, Ian D. [1 ,2 ]
Mankouri, Hocine W. [1 ,2 ]
机构
[1] Univ Copenhagen, Dept Cellular & Mol Med, Ctr Chromosome Stabil, DK-2200 Copenhagen, Denmark
[2] Univ Copenhagen, Dept Cellular & Mol Med, Ctr Hlth Aging, DK-2200 Copenhagen, Denmark
[3] Univ Copenhagen, Fac Hlth & Med Sci, Novo Nordisk Fdn, Ctr Prot Res, DK-2200 Copenhagen, Denmark
基金
欧洲研究理事会; 新加坡国家研究基金会;
关键词
RecQ helicase; DNA replication stress; genome stability; mutagenesis; recombination; HOMOLOGOUS RECOMBINATION; SACCHAROMYCES-CEREVISIAE; SOMATIC MUTATIONS; STRESS; GENES; SGS1; MECHANISMS; SCREEN; CANCER; BLM;
D O I
10.1073/pnas.1706640114
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
Proliferating cells acquire genome alterations during the act of DNA replication. This leads to mutation accumulation and somatic cell mosaicism in multicellular organisms, and is also implicated as an underlying cause of aging and tumorigenesis. The molecular mechanisms of DNA replication-associated genome rearrangements are poorly understood, largely due to methodological difficulties in analyzing specific replication forks in vivo. To provide an insight into this process, we analyzed themutagenic consequences of replication fork stalling at a single, site-specific replication barrier (the Escherichia coli Tus/Ter complex) engineered into the yeast genome. We demonstrate that transient stalling at this barrier induces a distinct pattern of genome rearrangements in the newly replicated region behind the stalled fork, which primarily consist of localized losses and duplications of DNA sequences. These genetic alterations arise through the aberrant repair of a single-stranded DNA gap, in a process that is dependent on Exo1- and Shu1-dependent homologous recombination repair (HRR). Furthermore, aberrant processing of HRR intermediates, and elevated HRR-associated mutagenesis, is detectable in a yeast model of the human cancer predisposition disorder, Bloom's syndrome. Our data reveal a mechanism by which cellular responses to stalled replication forks can actively generate genomic alterations and genetic diversity in normal proliferating cells.
引用
收藏
页码:9665 / 9670
页数:6
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