Substrate-selective repair and restart of replication forks by DNA translocases.

Substrate-selective repair and restart of replication forks by DNA translocases.
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DOI:
10.1016/j.celrep.2013.05.002
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发表时间:
2013-06-27
期刊:
影响因子:
8.8
通讯作者:
Cortez D
Cortez D
中科院分区:
生物学1区
文献类型:
--
作者:
Bétous R;Couch FB;Mason AC;Eichman BF;Manosas M;Cortez D

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停滞的复制分叉是遗传不稳定的根源。多种叉子重塑酶被招募来停止叉子,但它们是如何促进叉子重新启动的,人们知之甚少。通过集合生化分析和单分子磁镊子研究,我们发现SMARCAL1分支迁移和DNA退火活动是由单链DNA结合蛋白RPA指导的,以选择性地回归由前导链聚合酶阻塞引起的停滞复制叉,并恢复具有滞后链间隙的正常复制叉。我们揭示了RPA增强SMARCAL1底物偏好的分子机制。大肠杆菌RecG在存在大肠杆菌SSB时的作用与SMARCAL1相似,而高度相关的人类蛋白ZRANB3具有不同的底物偏好。我们的研究结果确定了SMARCAL1在叉子修复中的重要底物,表明RecG和SMARCAL1是功能同源物,并提供了这些DNA转位修复叉子的综合模型。
Stalled replication forks are sources of genetic instability. Multiple fork remodeling enzymes are recruited to stalled forks, but how they work to promote fork restart is poorly understood. By combining ensemble biochemical assays and single molecule studies with magnetic tweezers, we show that SMARCAL1 branch migration and DNA annealing activities are directed by the single-stranded DNA binding protein RPA to selectively regress stalled replication forks caused by blockage to the leading-strand polymerase and to restore normal replication forks with a lagging-strand gap. We unveil the molecular mechanisms by which RPA enforces SMARCAL1 substrate preference. E. coli RecG acts similarly to SMARCAL1 in the presence of E. coli SSB, whereas the highly related human protein ZRANB3 has different substrate preferences. Our findings identify the important substrates of SMARCAL1 in fork repair, suggest that RecG and SMARCAL1 are functional orthologues, and provide a comprehensive model of fork repair by these DNA translocases.
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