Escherichia coli Rep DNA helicase and error-prone DNA polymerase IV interact physically and functionally.

Escherichia coli Rep DNA helicase and error-prone DNA polymerase IV interact physically and functionally.
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DOI:
10.1111/j.1365-2958.2011.07590.x
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发表时间:
2011-04
影响因子:
3.6
通讯作者:
Foster PL
Foster PL
中科院分区:
生物学2区
文献类型:
--
作者:
Sladewski TE;Hetrick KM;Foster PL

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大肠杆菌DNA聚合酶IV由dinB基因编码,是特殊DNA聚合酶Y家族的成员。Pol IV能够合成过去的DNA损伤,并可能有助于重新启动停滞的复制分叉。然而,Pol IV是容易出错的,有助于DNA损伤诱导和应激诱导(适应性)突变。在这里,我们证明了Pol IV在体外与Rep DNA解旋酶相互作用,并且这种相互作用增强了Rep解旋酶的活性。此外,Pol IV聚合酶活性是通过与Rep相互作用而激活的,而Pol IV β钳结合基序似乎是这种激活所必需的。然而,Rep的解旋酶活性和结合DNA的能力都不需要它来刺激Pol IV的聚合酶活性。Rep和Pol IV之间的相互作用在体内具有重要的生物学意义,因为Rep增强了Pol IV在静止期细胞中的诱变活性。这些数据表明Rep在Pol iv依赖性适应性突变中的新作用。这种功能性相互作用也为细胞如何控制或靶向Pol IV的致突变活性提供了新的见解。
Escherichia coli DNA polymerase IV, encoded by the dinB gene, is a member of the Y family of specialized DNA polymerases. Pol IV is capable of synthesizing past DNA lesions and may help to restart stalled replication forks. However, Pol IV is error-prone, contributing to both DNA damage-induced and stress-induced (adaptive) mutations. Here we demonstrate that Pol IV interacts in vitro with Rep DNA helicase and that this interaction enhances Rep’s helicase activity. In addition, Pol IV polymerase activity is stimulated by interacting with Rep, and Pol IV β clamp-binding motif appears to be required for this stimulation. However, neither Rep’s helicase activity nor its ability to bind DNA is required for it to stimulate Pol IV’s polymerase activity. The interaction between Rep and Pol IV is biologically significant in vivo as Rep enhances Pol IV’s mutagenic activity in stationary-phase cells. These data indicate a new role for Rep in contributing to Pol IV-dependent adaptive mutation. This functional interaction also provides new insight into how the cell might control or target Pol IV’s mutagenic activity.
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