Reconstitution of SPO11-dependent double-strand break formation

被引：3

作者：

Zheng, Zhi ^{[1
,2
]}

Zheng, Lyuqin ^{[2
]}

Arter, Meret ^{[2
]}

Liu, Kaixian ^{[2
]}

Yamada, Shintaro ^{[2
,3
,4
]}

Ontoso, David ^{[2
,5
]}

Kim, Soonjoung ^{[2
,6
]}

Keeney, Scott ^{[1
,2
,5
]}

机构：

[1] Mem Sloan Kettering Canc Ctr, Louis V Gerstner Jr Grad Sch Biomed Sci, New York, NY 10021 USA

[2] Mem Sloan Kettering Canc Ctr, Mol Biol Program, New York, NY 10065 USA

[3] Kyoto Univ, HAKUBI Ctr Adv Res, Grad Sch Med, Kyoto, Japan

[4] Kyoto Univ, Grad Sch Med, Dept Aging Sci & Med, Kyoto, Japan

[5] Mem Sloan Kettering Canc Ctr, Howard Hughes Med Inst, New York, NY 10065 USA

[6] Yonsei Univ, Coll Med, Inst Immunol & Immunol Dis, Dept Microbiol & Immunol, Seoul, South Korea

来源：

NATURE | 2025年

关键词：

DNA TOPOISOMERASE-VI; MEIOTIC RECOMBINATION; MEIOSIS; SPO11; INITIATION; COMPLEXES; DYNAMICS; DISTINCT;

D O I：

10.1038/s41586-025-08601-2

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：

Meiotic recombination starts with SPO11 generation of DNA double-strand breaks (DSBs)1. SPO11 is critical for meiosis in most species, but it generates dangerous DSBs with mutagenic2 and gametocidal3 potential. Cells must therefore utilize the beneficial functions of SPO11 while minimizing its risks4-how they do so remains poorly understood. Here we report reconstitution of DNA cleavage in vitro with purified recombinant mouse SPO11 bound to TOP6BL. SPO11-TOP6BL complexes are monomeric (1:1) in solution and bind tightly to DNA, but dimeric (2:2) assemblies cleave DNA to form covalent 5 ' attachments that require SPO11 active-site residues, divalent metal ions and SPO11 dimerization. SPO11 can also reseal DNA that it has nicked. Structure modelling with AlphaFold 3 suggests that DNA is bent prior to cleavage5. In vitro cleavage displays a sequence bias that partially explains DSB site preferences in vivo. Cleavage is inefficient on complex DNA substrates, partly because SPO11 is readily trapped in DSB-incompetent (presumably monomeric) binding states that exchange slowly. However, cleavage is improved with substrates that favour dimer assembly or by artificially dimerizing SPO11. Our results inform a model in which intrinsically weak dimerization restrains SPO11 activity in vivo, making it exquisitely dependent on accessory proteins that focus and control DSB formation.

引用

页码：784 / 791

页数：25

共 66 条

[1] Lam I., Keeney S., Mechanism and regulation of meiotic recombination initiation, Cold Spring Harb. Perspect. Biol, 7, (2014)
[2] Hinch R., Donnelly P., Hinch A.G., Meiotic DNA breaks drive multifaceted mutagenesis in the human germ line, Science, 382, (2023)
[3] Handel M.A., Schimenti J.C., Genetics of mammalian meiosis: regulation, dynamics and impact on fertility, Nat. Rev. Genet, 11, pp. 124-136, (2010)
[4] Keeney S., Lange J., Mohibullah N., Self-organization of meiotic recombination initiation: general principles and molecular pathways, Annu. Rev. Genet, 48, pp. 187-214, (2014)
[5] Abramson J., Et al., Accurate structure prediction of biomolecular interactions with AlphaFold 3, Nature, 630, pp. 493-500, (2024)
[6] Hunter N., Meiotic recombination: the essence of heredity, Cold Spring Harb. Perspect. Biol, 7, pp. 361-395, (2015)
[7] Zickler D., Kleckner N., Meiosis: dances between homologs, Annu. Rev. Genet, 57, pp. 1-63, (2023)
[8] Jiao Y., Et al., A TOP6BL mutation abolishes meiotic DNA double-strand break formation and causes human infertility, Sci. Bull, 65, pp. 2120-2129, (2020)
[9] Robert T., Et al., The TopoVIB-like protein family is required for meiotic DNA double-strand break formation, Science, 351, pp. 943-949, (2016)
[10] Baudat F., Manova K., Yuen J.P., Jasin M., Keeney S., Chromosome synapsis defects and sexually dimorphic meiotic progression in mice lacking Spo11, Mol. Cell, 6, pp. 989-998, (2000)

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