A new protein complex promoting the assembly of Rad51 filaments.

A new protein complex promoting the assembly of Rad51 filaments.
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DOI:
10.1038/ncomms2678
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发表时间:
2013
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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在同源重组过程中,真核生物的RecA同源物Rad51在单链DNA上组装成核蛋白丝,催化与同源模板的同源配对和DNA链交换。Rad51核蛋白丝是高度动态的,并通过包括Rad51介质在内的各种辅助蛋白的协调作用进行调节。在这里,我们确定了一个新的Rad51中介复合物。PCSS复合体包括出芽酵母Psy3、Csm2、Shu1和Shu2蛋白,结合重组位点,是Rad51减数分裂过程中组装和功能所必需的。在异四聚体中,Psy3-Csm2构成一个具有dna结合活性的核心亚复合物。在体外,纯化的Psy3-Csm2可以独立于核苷酸辅助因子稳定rad51 -单链DNA复合体。我们的高分辨率晶体结构推断了Rad51稳定的机制,揭示了Psy3-Csm2是Rad51-二聚体的结构模拟物,Rad51-灯丝的基本单元。总之,这些结果揭示了这类Rad51介质的一种新的分子机制,其中包括人类Rad51同源物。
During homologous recombination, eukaryotic RecA homologue Rad51 assembles into a nucleoprotein filament on single-stranded DNA to catalyse homologous pairing and DNA-strand exchange with a homologous template. Rad51 nucleoprotein filaments are highly dynamic and regulated via the coordinated actions of various accessory proteins including Rad51 mediators. Here, we identify a new Rad51 mediator complex. The PCSS complex, comprising budding yeast Psy3, Csm2, Shu1 and Shu2 proteins, binds to recombination sites and is required for Rad51 assembly and function during meiosis. Within the heterotetramer, Psy3-Csm2 constitutes a core sub-complex with DNA-binding activity. In vitro, purified Psy3-Csm2 stabilizes the Rad51–single-stranded DNA complex independently of nucleotide cofactor. The mechanism of Rad51 stabilization is inferred by our high-resolution crystal structure, which reveals Psy3-Csm2 to be a structural mimic of the Rad51-dimer, a fundamental unit of the Rad51-filament. Together, these results reveal a novel molecular mechanism for this class of Rad51-mediators, which includes the human Rad51 paralogues.
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