Fork sensing and strand switching control antagonistic activities of RecQ helicases.

Fork sensing and strand switching control antagonistic activities of RecQ helicases.
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DOI:
10.1038/ncomms3024
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发表时间:
2013
影响因子:
16.6
通讯作者:
Seidel R
Seidel R
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Klaue D;Kobbe D;Kemmerich F;Kozikowska A;Puchta H;Seidel R

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RecQ解旋酶在复制过程中维持基因组稳定性和通过同源重组控制双链断裂修复中具有重要作用。关于高等真核生物中发现的不同RecQ解旋酶如何实现其专门和部分相反的功能,人们知之甚少。在这里,我们调查的RecQ解旋酶从拟南芥,AtRECQ2和AtRECQ3的DNA解旋在单分子水平上使用磁镊。虽然AtRECQ2主要以高度重复的方式解旋分叉的DNA底物,但AtRECQ3更喜欢倒带,即关闭预先打开的DNA叉。对于这两种酶,这一过程是由频繁的链开关和主动传感解旋叉控制。链朝向解旋或朝向重绕的相对程度决定了酶的主要方向。我们的研究结果提供了一个简单的解释如何不同的生物活性可以实现相当相似的RecQ家族成员。 RecQ解旋酶是在DNA修复级联中维持基因组稳定性中发挥核心作用的酶。Klaue等人表明,来自拟南芥的RecQ2和RecQ3使用频繁的链转换机制,分别通过解旋和重绕分叉的DNA底物来加工DNA。
RecQ helicases have essential roles in maintaining genome stability during replication and in controlling double-strand break repair by homologous recombination. Little is known about how the different RecQ helicases found in higher eukaryotes achieve their specialized and partially opposing functions. Here, we investigate the DNA unwinding of RecQ helicases from Arabidopsis thaliana, AtRECQ2 and AtRECQ3 at the single-molecule level using magnetic tweezers. Although AtRECQ2 predominantly unwinds forked DNA substrates in a highly repetitive fashion, AtRECQ3 prefers to rewind, that is, to close preopened DNA forks. For both enzymes, this process is controlled by frequent strand switches and active sensing of the unwinding fork. The relative extent of the strand switches towards unwinding or towards rewinding determines the predominant direction of the enzyme. Our results provide a simple explanation for how different biological activities can be achieved by rather similar members of the RecQ family. RecQ helicases are enzymes that play a central role in maintaining genome stability in the DNA repair cascade. Klaue et al. show that RecQ2 and RecQ3 from Arabidopsis thaliana process DNA by, respectively, unwinding and rewinding forked DNA substrates, using a frequent strand switching mechanism.
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