A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis

A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis Rita Tewari, Ryuji Yanase, Molly Hair, Mohammad Zeeshan, David J. P. Ferguson, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8180258/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Meiosis is a conserved yet evolutionarily varied process underpinning sexual reproduction in eukaryotes. In the malaria parasite Plasmodium , meiosis is unconventional: it occurs immediately after fertilisation (post-zygotic) and must be coordinated with the transformation of the zygote into a motile ookinete. The mechanisms synchronising these meiotic and morphogenetic programmes remain unknow. Here, we identify the Plasmodium berghei NIMA-related kinase, NEK4 as a key regulator that couples meiotic initiation with zygote morphogenesis. Using ultrastructure expansion microscopy, we show that NEK4 accumulates at the microtubule-organising centre (MTOC) and the apical polar complex (APC) shortly after fertilisation, preceding the assembly of perinuclear and cortical microtubules. We reveal that Plasmodium zygotes undergo a nuclear migration driven by the MTOC, analogous to the meiotic nuclear movement in fission yeast. Deletion of nek4 results in complete developmental arrest: MTOC duplication and microtubule formation are blocked, chromatin remains uncondensed, and nuclear migration and cell polarity fail to establish. Transcriptomic and phosphoproteomic analyses reveal that NEK4 absence causes a collapse in transcriptional and phosphoregulatory networks governing meiosis and cytoskeletal organisation, leading to reduced expression and phosphorylation of important players, including HOP1, REC8, and AP2-O. These findings establish NEK4 as a key regulator driving meiotic entry and zygote maturation. Biological sciences/Microbiology/Parasitology/Parasite biology Biological sciences/Cell biology/Cell division/Meiosis Biological sciences/Cell biology/Cytoskeleton/Microtubules Plasmodium meiosis NIMA-related kinase malaria zygote ookinete MTOC microtubules chromosome Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Supplementarytables114.xlsx Supplementary tables 1-14 NEK4SupplementaryFigurescompressed.pdf Supplementary figures 1-5 SupplementaryVideo2.mp4 Supplementary Video 2 SupplementaryVideo1.mp4 Supplementary Video 1 SupplementaryVideo4.mp4 Supplementary Video 4 SupplementaryVideo3.mp4 Supplementary Video 3 Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8180258","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":555958085,"identity":"d95dba22-fa93-45c0-976f-902eb20aa65f","order_by":0,"name":"Rita Tewari","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0003-3943-1847","institution":"University of Nottingham","correspondingAuthor":true,"prefix":"","firstName":"Rita","middleName":"","lastName":"Tewari","suffix":""},{"id":555958086,"identity":"4b0acdab-8ed3-405c-883c-a7ce29c2cd46","order_by":1,"name":"Ryuji Yanase","email":"","orcid":"https://orcid.org/0000-0001-9419-398X","institution":"University of Nottingham","correspondingAuthor":false,"prefix":"","firstName":"Ryuji","middleName":"","lastName":"Yanase","suffix":""},{"id":555958087,"identity":"014e848a-d459-4d74-b11b-42d2882716a0","order_by":2,"name":"Molly Hair","email":"","orcid":"","institution":"Oxford Brookes University","correspondingAuthor":false,"prefix":"","firstName":"Molly","middleName":"","lastName":"Hair","suffix":""},{"id":555958088,"identity":"3c73cfe1-e7ad-48ec-a440-b768feb119ac","order_by":3,"name":"Mohammad Zeeshan","email":"","orcid":"https://orcid.org/0000-0002-6185-403X","institution":"International Centre for Genetic Engineering and Biotechnology (ICGEB)","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Zeeshan","suffix":""},{"id":555958089,"identity":"64d7ee4e-6f4d-4a89-bd99-6143442708ed","order_by":4,"name":"David J. 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meiosis","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Plasmodium, meiosis, NIMA-related kinase, malaria, zygote, ookinete, MTOC, microtubules, chromosome","lastPublishedDoi":"10.21203/rs.3.rs-8180258/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8180258/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Meiosis is a conserved yet evolutionarily varied process underpinning sexual reproduction in eukaryotes. In the malaria parasite \u003ci\u003ePlasmodium\u003c/i\u003e, meiosis is unconventional: it occurs immediately after fertilisation (post-zygotic) and must be coordinated with the transformation of the zygote into a motile ookinete. The mechanisms synchronising these meiotic and morphogenetic programmes remain unknow. Here, we identify the \u003ci\u003ePlasmodium berghei\u003c/i\u003e NIMA-related kinase, NEK4 as a key regulator that couples meiotic initiation with zygote morphogenesis. Using ultrastructure expansion microscopy, we show that NEK4 accumulates at the microtubule-organising centre (MTOC) and the apical polar complex (APC) shortly after fertilisation, preceding the assembly of perinuclear and cortical microtubules. We reveal that \u003ci\u003ePlasmodium\u003c/i\u003e zygotes undergo a nuclear migration driven by the MTOC, analogous to the meiotic nuclear movement in fission yeast. Deletion of \u003ci\u003enek4\u003c/i\u003e results in complete developmental arrest: MTOC duplication and microtubule formation are blocked, chromatin remains uncondensed, and nuclear migration and cell polarity fail to establish. Transcriptomic and phosphoproteomic analyses reveal that NEK4 absence causes a collapse in transcriptional and phosphoregulatory networks governing meiosis and cytoskeletal organisation, leading to reduced expression and phosphorylation of important players, including HOP1, REC8, and AP2-O. These findings establish NEK4 as a key regulator driving meiotic entry and zygote maturation.","manuscriptTitle":"A divergent Plasmodium NEK4 acts as a key regulator driving the early events of meiosis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-09 04:02:14","doi":"10.21203/rs.3.rs-8180258/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"4930e2db-1ded-410f-ba0f-c8c128546a04","owner":[],"postedDate":"December 9th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":59175399,"name":"Biological sciences/Microbiology/Parasitology/Parasite biology"},{"id":59175400,"name":"Biological sciences/Cell biology/Cell division/Meiosis"},{"id":59175401,"name":"Biological sciences/Cell biology/Cytoskeleton/Microtubules"}],"tags":[],"updatedAt":"2026-05-05T09:15:38+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-09 04:02:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8180258","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8180258","identity":"rs-8180258","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}