Structural and functional characterization of the Spo11 core complex

IF 16.8 1区 生物学
Corentin Claeys Bouuaert, Sam E. Tischfield, Stephen Pu, Eleni P. Mimitou, E. Arias-Palomo, J. Berger, S. Keeney
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引用次数: 27

Abstract

Spo11, which makes DNA double-strand breaks (DSBs) that are essential for meiotic recombination, has long been recalcitrant to biochemical study. We provide molecular analysis of Saccharomyces cerevisiae Spo11 purified with partners Rec102, Rec104 and Ski8. Rec102 and Rec104 jointly resemble the B subunit of archaeal topoisomerase VI, with Rec104 occupying a position similar to the Top6B GHKL-type ATPase domain. Unexpectedly, the Spo11 complex is monomeric (1:1:1:1 stoichiometry), consistent with dimerization controlling DSB formation. Reconstitution of DNA binding reveals topoisomerase-like preferences for duplex–duplex junctions and bent DNA. Spo11 also binds noncovalently but with high affinity to DNA ends mimicking cleavage products, suggesting a mechanism to cap DSB ends. Mutations that reduce DNA binding in vitro attenuate DSB formation, alter DSB processing and reshape the DSB landscape in vivo. Our data reveal structural and functional similarities between the Spo11 core complex and Topo VI, but also highlight differences reflecting their distinct biological roles. Biochemical and structural characterization of the meiotic DSB core complex of budding yeast reveals molecular architecture and DNA-binding properties similar to those of ancestral Topo VI.
Spo11核配合物的结构与功能表征
Spo11产生DNA双链断裂(DSBs),这对减数分裂重组至关重要,长期以来一直难以进行生化研究。我们提供了与伴侣Rec102、Rec104和Ski8纯化的酿酒酵母Spo11的分子分析。Rec102和Rec104共同类似于古菌拓扑异构酶VI的B亚基,Rec104占据类似于Top6B GHKL型ATP酶结构域的位置。出乎意料的是,Spo11络合物是单体的(化学计量为1:1:1:1),与控制DSB形成的二聚化一致。DNA结合的重组揭示了拓扑异构酶对双链连接和弯曲DNA的偏好。Spo11也以非共价结合,但与模拟切割产物的DNA末端具有高亲和力,这表明了一种覆盖DSB末端的机制。在体外减少DNA结合的突变减弱了DSB的形成,改变了DSB加工,并在体内重塑了DSB景观。我们的数据揭示了Spo11核心复合体和Topo VI在结构和功能上的相似性,但也突出了反映其不同生物学作用的差异。出芽酵母减数分裂DSB核心复合物的生化和结构表征揭示了与祖先Topo VI相似的分子结构和DNA结合特性。
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来源期刊
Nature Structural &Molecular Biology
Nature Structural &Molecular Biology 生物-生化与分子生物学
自引率
1.80%
发文量
160
期刊介绍: Nature Structural & Molecular Biology is a monthly journal that focuses on the functional and mechanistic understanding of how molecular components in a biological process work together. It serves as an integrated forum for structural and molecular studies. The journal places a strong emphasis on the functional and mechanistic understanding of how molecular components in a biological process work together. Some specific areas of interest include the structure and function of proteins, nucleic acids, and other macromolecules, DNA replication, repair and recombination, transcription, regulation of transcription and translation, protein folding, processing and degradation, signal transduction, and intracellular signaling.
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