{"doi":"10.1038/s41586-025-08601-2","title":"Reconstitution of SPO11-dependent double-strand break formation","abstract":"<jats:title>Abstract</jats:title>\n                  <jats:p>\n                    Meiotic recombination starts with SPO11 generation of DNA double-strand breaks (DSBs)\n                    <jats:sup>1</jats:sup>\n                    . SPO11 is critical for meiosis in most species, but it generates dangerous DSBs with mutagenic\n                    <jats:sup>2</jats:sup>\n                    and gametocidal\n                    <jats:sup>3</jats:sup>\n                    potential. Cells must therefore utilize the beneficial functions of SPO11 while minimizing its risks\n                    <jats:sup>4</jats:sup>\n                    —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 cleavage\n                    <jats:sup>5</jats:sup>\n                    . 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.\n                  </jats:p>","journal":"Nature","year":2025,"id":646690,"datarank":0.5101796072493234,"base_score":3.4011973816621555,"endowment":3.4011973816621555,"self_citation_contribution":0.5101796072493234,"citation_network_contribution":0.0,"self_endowment_contribution":0.5101796072493234,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":29,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1684578,"name":"Lyuqin Zheng","orcid":null,"position":1,"is_corresponding":false},{"id":1162415,"name":"Meret Arter","orcid":"0000-0002-1734-4622","position":2,"is_corresponding":false},{"id":497871,"name":"Kaixian Liu","orcid":"0000-0001-9711-6509","position":3,"is_corresponding":false},{"id":266093,"name":"Shintaro Yamada","orcid":"0000-0002-4882-0871","position":4,"is_corresponding":false},{"id":1338701,"name":"David Ontoso","orcid":"0000-0003-3831-178X","position":5,"is_corresponding":false},{"id":1084787,"name":"Soonjoung Kim","orcid":"0009-0003-0029-4715","position":6,"is_corresponding":false},{"id":254578,"name":"Scott Keeney","orcid":"0000-0002-1283-6417","position":7,"is_corresponding":false},{"id":806406,"name":"Zhi Zheng","orcid":"0000-0003-0867-7334","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Reconstitution of SPO11-dependent double-strand break formation","abstract":"<jats:title>Abstract</jats:title>\n                  <jats:p>\n                    Meiotic recombination starts with SPO11 generation of DNA double-strand breaks (DSBs)\n                    <jats:sup>1</jats:sup>\n                    . SPO11 is critical for meiosis in most species, but it generates dangerous DSBs with mutagenic\n                    <jats:sup>2</jats:sup>\n                    and gametocidal\n                    <jats:sup>3</jats:sup>\n                    potential. Cells must therefore utilize the beneficial functions of SPO11 while minimizing its risks\n                    <jats:sup>4</jats:sup>\n                    —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 cleavage\n                    <jats:sup>5</jats:sup>\n                    . 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.\n                  </jats:p>","is_dataset_classified":null,"base_score":3.4011973816621555,"endowment":3.4011973816621555,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"39972129","pmcid":"PMC11922745","openalex_id":"https://openalex.org/W4407722125","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"R35 GM118092","title":null},{"funder_name":"NCI NIH HHS","grant_id":"P30 CA008748","title":null},{"funder_name":"NICHD NIH HHS","grant_id":"R01 HD110120","title":null},{"funder_name":"National Institutes of Health","grant_id":"2P30CA008748-43","title":"MOUSE GENETICS"},{"funder_name":"National Institutes of Health","grant_id":"5R01HD110120-04","title":"Structural and functional principles underlying germline genome transmission"},{"funder_name":"National Institutes of Health","grant_id":"2R35GM118092-06","title":"Mechanism and regulation of meiotic recombination"}],"total_grants":6,"fwci":11.0982,"citation_percentile":0.98998301,"influential_citations":0,"citation_trend":[{"year":2022,"count":1},{"year":2025,"count":13},{"year":2026,"count":15}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.1038/s41586-025-08601-2","host_type":"journal"},{"url":"https://doi.org/10.1038/s41586-025-08601-2","host_type":"publisher"},{"url":"https://www.nature.com/articles/s41586-025-08601-2.pdf","host_type":"publisher"},{"url":"https://www.nature.com/articles/s41586-025-08601-2","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/39972129","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/11922745","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC11922745","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC11922745?pdf=render","host_type":"Europe_PMC"},{"url":"https://doi.org/10.1101/2024.11.20.624382","host_type":""},{"url":"https://pubmed.ncbi.nlm.nih.gov/39605552","host_type":""},{"url":"http://dx.doi.org/10.1101/2024.11.20.624382","host_type":""},{"url":"http://dx.doi.org/10.1038/s41586-025-08601-2","host_type":""}],"fields_of_study":["DNA Repair Mechanisms","DNA and Nucleic Acid Chemistry","Carcinogens and Genotoxicity Assessment","0301 basic medicine","03 medical and health sciences"],"mesh_terms":["Meiotic Recombination Protein SPO11","Animals","DNA","Endodeoxyribonucleases","Meiosis","Models, Molecular","Protein Binding","Recombinant Proteins","Substrate Specificity","Catalytic Domain","Mice","DNA Cleavage","DNA Breaks, Double-Stranded","Protein Multimerization"],"keywords":["Cleavage (geology)","DNA","Recombinant DNA","Homologous recombination","Biophysics","Holliday junction","Chemistry","Cell biology","In vitro","Divalent","Biology","Biochemistry","Gene","Models, Molecular","Endodeoxyribonucleases","Article","Mice","Meiosis","Catalytic Domain","Animals","DNA Breaks, Double-Stranded","Meiotic Recombination Protein SPO11","Protein Multimerization","DNA Cleavage","Protein Binding"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life in Land"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"geo"},{"name":"uniprot"},{"name":"pdb"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-09T14:29:33.961680Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}