{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T18:14:29Z","timestamp":1778004869235,"version":"3.51.4"},"update-to":[{"DOI":"10.1371\/journal.ppat.1014181","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T00:00:00Z","timestamp":1777939200000}}],"reference-count":32,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"DOI":"10.13039\/100006492","name":"Division of Intramural Research, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["Not applicable"],"award-info":[{"award-number":["Not applicable"]}],"id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    Genetic exchange in\n                    <jats:italic>Leishmania<\/jats:italic>\n                    is established, yet the molecular mechanisms enabling hybrid formation in sand flies remain poorly defined. In\n                    <jats:italic>Leishmania<\/jats:italic>\n                    , as in plants and several protists, two paralogs of the conserved meiotic endonuclease SPO11 are present, but their contribution to hybridization is unknown. Here, we dissect the roles of SPO11\u20131 and SPO11\u20132 during in vivo sand fly infections using targeted gene deletions, catalytically-dead mutants, expression analyses, and genome-wide characterization of hybrid progeny. We show that both SPO11 paralogs are essential for efficient hybridization: deletion of either paralog in both parents abolishes hybrid recovery, and catalytic inactivation fails to rescue mating. When only one parent lacks SPO11\u20131 or SPO11\u20132, hybrid formation is reduced in a strain-dependent manner, revealing asymmetric requirements for each paralog. Genomic analysis of hybrids from SPO11-deficient crosses reveals polyploidy and altered parental genome contributions, including unbalanced and near-balanced triploid configurations, indicating disrupted reductional processes. Together, these results establish SPO11-dependent DNA break formation as a core requirement for\n                    <jats:italic>Leishmania<\/jats:italic>\n                    hybridization and define distinct, strain-specific roles for the two SPO11 paralogs.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1014181","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T17:52:13Z","timestamp":1777485133000},"page":"e1014181","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Cryptic sex in Leishmania depends on SPO11 paralogs"],"prefix":"10.1371","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8983-3487","authenticated-orcid":true,"given":"Carolina M. C.","family":"Catta-Preta","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vitor Luiz","family":"da Silva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudio","family":"Meneses","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kashinath","family":"Ghosh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David L.","family":"Sacks","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"issue":"3","key":"ppat.1014181.ref001","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/S0092-8674(00)81876-0","article-title":"Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family","volume":"88","author":"S Keeney","year":"1997","journal-title":"Cell"},{"issue":"2","key":"ppat.1014181.ref002","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1093\/genetics\/110.2.187","article-title":"The role of the SPO11 gene in meiotic recombination in yeast","volume":"110","author":"S Klapholz","year":"1985","journal-title":"Genetics"},{"issue":"18","key":"ppat.1014181.ref003","doi-asserted-by":"crossref","first-page":"2932","DOI":"10.1101\/gad.12.18.2932","article-title":"mei-W68 in Drosophila melanogaster encodes a Spo11 homolog: evidence that the mechanism for initiating meiotic recombination is conserved","volume":"12","author":"KS McKim","year":"1998","journal-title":"Genes Dev"},{"issue":"3","key":"ppat.1014181.ref004","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/S0092-8674(00)81481-6","article-title":"Meiotic recombination in C. elegans initiates by a conserved mechanism and is dispensable for homologous chromosome synapsis","volume":"94","author":"AF Dernburg","year":"1998","journal-title":"Cell"},{"issue":"3","key":"ppat.1014181.ref005","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1093\/emboj\/20.3.589","article-title":"AtSPO11-1 is necessary for efficient meiotic recombination in plants","volume":"20","author":"M Grelon","year":"2001","journal-title":"EMBO J"},{"issue":"5","key":"ppat.1014181.ref006","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1016\/S1097-2765(00)00097-6","article-title":"The mouse Spo11 gene is required for meiotic chromosome synapsis","volume":"6","author":"PJ Romanienko","year":"2000","journal-title":"Mol Cell"},{"issue":"12","key":"ppat.1014181.ref007","doi-asserted-by":"crossref","first-page":"2827","DOI":"10.1093\/molbev\/msm217","article-title":"Protist homologs of the meiotic Spo11 gene and topoisomerase VI reveal an evolutionary history of gene duplication and lineage-specific loss","volume":"24","author":"S-B Malik","year":"2007","journal-title":"Mol Biol Evol"},{"issue":"2","key":"ppat.1014181.ref008","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1111\/j.1365-313X.2006.02867.x","article-title":"Arabidopsis SPO11-2 functions with SPO11-1 in meiotic recombination","volume":"48","author":"NJ Stacey","year":"2006","journal-title":"Plant J"},{"issue":"10","key":"ppat.1014181.ref009","doi-asserted-by":"crossref","first-page":"3090","DOI":"10.1105\/tpc.107.054817","article-title":"The catalytically active tyrosine residues of both SPO11-1 and SPO11-2 are required for meiotic double-strand break induction in Arabidopsis","volume":"19","author":"F Hartung","year":"2007","journal-title":"Plant Cell"},{"key":"ppat.1014181.ref010","article-title":"Sexual reproduction and genetic exchange in parasitic protists","author":"GD Weedall","year":"2015","journal-title":"Parasitology"},{"issue":"3","key":"ppat.1014181.ref011","doi-asserted-by":"crossref","DOI":"10.1128\/spectrum.04980-22","article-title":"Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation","volume":"11","author":"P Singh","year":"2023","journal-title":"Microbiol Spectr"},{"issue":"9","key":"ppat.1014181.ref012","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0074465","article-title":"Homologous recombination occurs in Entamoeba and is enhanced during growth stress and stage conversion","volume":"8","author":"N Singh","year":"2013","journal-title":"PLoS One"},{"issue":"2","key":"ppat.1014181.ref013","first-page":"375","article-title":"Amoebozoans Are Secretly but Ancestrally Sexual: Evidence for Sex Genes and Potential Novel Crossover Pathways in Diverse Groups of Amoebae","volume":"9","author":"YI Tekle","year":"2017","journal-title":"Genome Biol Evol"},{"issue":"5869","key":"ppat.1014181.ref014","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.1126\/science.1153752","article-title":"Evidence for karyogamy and exchange of genetic material in the binucleate intestinal parasite Giardia intestinalis","volume":"319","author":"MK Poxleitner","year":"2008","journal-title":"Science"},{"issue":"9","key":"ppat.1014181.ref015","doi-asserted-by":"crossref","first-page":"3671","DOI":"10.1073\/pnas.1019423108","article-title":"Identification of the meiotic life cycle stage of Trypanosoma brucei in the tsetse fly","volume":"108","author":"L Peacock","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"1","key":"ppat.1014181.ref016","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1146\/annurev-micro-050724-091231","article-title":"Genetic Exchange in Leishmania: Understanding the Cryptic Sexual Cycle","volume":"79","author":"CMC Catta-Preta","year":"2025","journal-title":"Annu Rev Microbiol"},{"issue":"5924","key":"ppat.1014181.ref017","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1126\/science.1169464","article-title":"Demonstration of genetic exchange during cyclical development of Leishmania in the sand fly vector","volume":"324","author":"NS Akopyants","year":"2009","journal-title":"Science"},{"issue":"5","key":"ppat.1014181.ref018","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0019851","article-title":"Visualisation of Leishmania donovani fluorescent hybrids during early stage development in the sand fly vector","volume":"6","author":"J Sadlova","year":"2011","journal-title":"PLoS One"},{"issue":"52","key":"ppat.1014181.ref019","article-title":"Single-cell atlas of Leishmania development in sandflies reveals the heterogeneity of transmitted parasites and their role in infection","volume":"121","author":"CMC Catta-Preta","year":"2024","journal-title":"Proc Natl Acad Sci U S 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Sci"},{"key":"ppat.1014181.ref029","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.73488","article-title":"Stress conditions promote Leishmania hybridization in vitro marked by expression of the ancestral gamete fusogen HAP2 as revealed by single-cell RNA-seq","volume":"11","author":"I Louradour","year":"2022","journal-title":"Elife"},{"issue":"1","key":"ppat.1014181.ref030","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1093\/nar\/gkab1203","article-title":"High throughput single-cell genome sequencing gives insights into the generation and evolution of mosaic aneuploidy in Leishmania donovani","volume":"50","author":"GH Negreira","year":"2022","journal-title":"Nucleic Acids Res"},{"key":"ppat.1014181.ref031","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1051\/parasite\/2017041","article-title":"Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae)","volume":"24","author":"P Lawyer","year":"2017","journal-title":"Parasite"},{"issue":"5","key":"ppat.1014181.ref032","doi-asserted-by":"crossref","first-page":"170095","DOI":"10.1098\/rsos.170095","article-title":"A CRISPR Cas9 high-throughput genome editing toolkit for kinetoplastids","volume":"4","author":"T Beneke","year":"2017","journal-title":"R Soc Open Sci"}],"updated-by":[{"DOI":"10.1371\/journal.ppat.1014181","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T00:00:00Z","timestamp":1777939200000}}],"container-title":["PLOS Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1014181","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T17:46:07Z","timestamp":1778003167000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1014181"}},"subtitle":[],"editor":[{"given":"\u00c1lvaro","family":"Acosta-Serrano","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,4,29]]},"references-count":32,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4,29]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.ppat.1014181","relation":{},"ISSN":["1553-7374"],"issn-type":[{"value":"1553-7374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,29]]}}}