SPO11 dimers are sufficient to catalyse DNA double-strand breaks in vitro

被引:2
作者
Oger, Cedric [1 ]
Bouuaert, Corentin Claeys [1 ]
机构
[1] Catholic Univ Louvain, Louvain Inst Biomol Sci & Technol, Louvain La Neuve, Belgium
基金
欧洲研究理事会;
关键词
MEIOTIC RECOMBINATION; TOPOISOMERASE-II; MEIOSIS; PRDM9; CONSERVATION; MECHANISM; BINDING; GENE;
D O I
10.1038/s41586-024-08574-8
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
SPO11 initiates meiotic recombination through the induction of programmed DNA double-strand breaks (DSBs)1,2, but this catalytic activity has never been reconstituted in vitro3,4. Here, using Mus musculus SPO11, we report a biochemical system that recapitulates all the hallmarks of meiotic DSB formation. We show that SPO11 catalyses break formation in the absence of any partners and remains covalently attached to the 5 ' broken strands. We find that target site selection by SPO11 is influenced by the sequence, bendability and topology of the DNA substrate, and provide evidence that SPO11 can reseal single-strand DNA breaks. In addition, we show that SPO11 is monomeric in solution and that cleavage requires dimerization for the reconstitution of two hybrid active sites. SPO11 and its partner TOP6BL form a 1:1 complex that catalyses DNA cleavage with an activity similar to that of SPO11 alone. However, this complex binds DNA ends with higher affinity, suggesting a potential role after cleavage. We propose a model in which additional partners of SPO11 required for DSB formation in vivo assemble biomolecular condensates that recruit SPO11-TOP6BL, enabling dimerization and cleavage. Our work establishes SPO11 dimerization as the fundamental mechanism that controls the induction of meiotic DSBs.
引用
收藏
页码:792 / 799
页数:23
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