Opposing roles of centrosomal and non-centrosomal branched F-actin networks in primary ciliogenesis

Opposing roles of centrosomal and non-centrosomal branched F-actin networks in primary ciliogenesis | 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 Opposing roles of centrosomal and non-centrosomal branched F-actin networks in primary ciliogenesis Pengli Zheng, Jianguo Chen, Junlin Teng, Yunjie Liu, Qiuchen Gu, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9467244/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 The primary cilium serves as a crucial signaling hub that integrates extracellular cues to coordinate development and maintain homeostasis, and its formation (ciliogenesis) is strictly regulated. Given the complex and spatially distinct organization of the actin filament cytoskeleton, the precise contributions of branched F-actin to ciliogenesis remain poorly understood. Here, we show that centrosomal branched actin filaments (c-BFA) facilitate ciliogenesis by directing the ARP2/3 nucleation‑promoting factor VCA to the centrosome. We identify LRCH2 as an actin‑binding protein that stabilizes c-BFA, thereby promoting ciliary vesicle docking at the mother centriole and enhancing ciliogenesis. In contrast, CCDC22, a known early endosome‑localized protein, regulates endosomal vesicle sorting by stabilizing non‑centrosomal branched actin filaments (nc-BFA), which consequently suppresses ciliary vesicle recruitment to the centrosome and inhibits ciliogenesis. Collectively, LRCH2 and CCDC22 antagonistically regulate branched actin networks at the centrosome and in the cytoplasm, thereby maintaining ciliogenesis homeostasis. Conditional knockout of Ccdc22 in mouse cerebellar granule cell precursors results in an increased proportion of ciliated cells and impairs balance function. Our findings highlight the distinct and opposing roles of c-BFA and nc-BFA in primary ciliogenesis. Biological sciences/Cell biology/Cytoskeleton/Cilia Biological sciences/Cell biology/Cytoskeleton/Actin ciliogenesis branched F-actin centrosome CCDC22 LRCH2 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryMovie1OpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.avi LRCH2 binds to and stablizes F-actin. SupplementaryOpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.pdf Supplementary figures SupplementaryMovie2OpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.mp4 Impaired balance beam performance in GCP-specific Ccdc22 knockout mice. 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-9467244","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":631143500,"identity":"1cde2e27-1e90-415b-8b09-4a7ff8579f51","order_by":0,"name":"Pengli Zheng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2ElEQVRIiWNgGAWjYDACCTBpIcfAA6LZiNciYUy6lsQGorXIz24+9vDrDon07TxnDBg+lB1m4J/dgF8L45xj6cayZyRyd/b2GDDOOHeYQeLOAfxamCVyzKQl2yRyN5znMWDmbTvMYCCRgF8Lm0T+N5CWdAOQlr/EaOGRyGGT/NgmkWBwtseAmZEYLRISaWbSjGckDHf2HCs42HMunUfiBgEt8jOSn0n+3GEjb86TvPHBjzJrOf4ZBLSAADNvAwODAZBxAORSwuqBgPEnVMsoGAWjYBSMAqwAANOyPShMs2wXAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3797-4023","institution":"Peking University","correspondingAuthor":true,"prefix":"","firstName":"Pengli","middleName":"","lastName":"Zheng","suffix":""},{"id":631143501,"identity":"70b2f06a-d556-4138-b095-9ea76ffc51a0","order_by":1,"name":"Jianguo Chen","email":"","orcid":"","institution":"Peking University","correspondingAuthor":false,"prefix":"","firstName":"Jianguo","middleName":"","lastName":"Chen","suffix":""},{"id":631143502,"identity":"f84c7a3d-9c29-4028-8838-5fc44605b6f1","order_by":2,"name":"Junlin Teng","email":"","orcid":"https://orcid.org/0000-0002-7611-0631","institution":"Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education and State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University","correspondingAuthor":false,"prefix":"","firstName":"Junlin","middleName":"","lastName":"Teng","suffix":""},{"id":631143503,"identity":"0e3a2b0f-e8a0-4ab0-861a-9ae964462484","order_by":3,"name":"Yunjie Liu","email":"","orcid":"","institution":"Peking University","correspondingAuthor":false,"prefix":"","firstName":"Yunjie","middleName":"","lastName":"Liu","suffix":""},{"id":631143504,"identity":"621accab-0e21-436b-a370-a842427d6de1","order_by":4,"name":"Qiuchen Gu","email":"","orcid":"","institution":"Peking University","correspondingAuthor":false,"prefix":"","firstName":"Qiuchen","middleName":"","lastName":"Gu","suffix":""},{"id":631143505,"identity":"626fb143-78db-4fb4-bfcc-708638cbe10d","order_by":5,"name":"Ning Huang","email":"","orcid":"https://orcid.org/0000-0002-8233-2572","institution":"Xi'an Jiaotong University","correspondingAuthor":false,"prefix":"","firstName":"Ning","middleName":"","lastName":"Huang","suffix":""}],"badges":[],"createdAt":"2026-04-20 05:40:51","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9467244/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9467244/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108183834,"identity":"1b28403e-37d3-4e9b-9e42-026e9eaffa43","added_by":"auto","created_at":"2026-04-30 09:02:55","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1591728,"visible":true,"origin":"","legend":"Article File","description":"","filename":"ManuscriptOpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9467244/v1_covered_42c9dd14-e4bc-4745-bb99-e651e9f268ee.pdf"},{"id":108067735,"identity":"f40dbbee-1b2f-4fdf-9bc8-d9f1db99a4c4","added_by":"auto","created_at":"2026-04-29 05:07:54","extension":"avi","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":559466,"visible":true,"origin":"","legend":"LRCH2 binds to and stablizes F-actin.","description":"","filename":"SupplementaryMovie1OpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.avi","url":"https://assets-eu.researchsquare.com/files/rs-9467244/v1/0ffc678e12ed80b7a4360770.avi"},{"id":108182003,"identity":"497ed2af-b208-43ff-b7d5-157f370e3c22","added_by":"auto","created_at":"2026-04-30 08:59:04","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":3174727,"visible":true,"origin":"","legend":"Supplementary figures","description":"","filename":"SupplementaryOpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9467244/v1/02e5f7a5dc08683ae7b3c4aa.pdf"},{"id":108067737,"identity":"0aa4fb92-cf1e-48b8-a299-05161b40a6d8","added_by":"auto","created_at":"2026-04-29 05:07:54","extension":"mp4","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":5738201,"visible":true,"origin":"","legend":"Impaired balance beam performance in GCP-specific Ccdc22 knockout mice.","description":"","filename":"SupplementaryMovie2OpposingRolesofCentrosomalandNonCentrosomalBranchedFactinNetworksinPrimaryCiliogenesis.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9467244/v1/ab8cbdfc30a1cd6643bfb236.mp4"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Opposing roles of centrosomal and non-centrosomal branched F-actin networks in primary ciliogenesis","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"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":"ciliogenesis, branched F-actin, centrosome, CCDC22, LRCH2","lastPublishedDoi":"10.21203/rs.3.rs-9467244/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9467244/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The primary cilium serves as a crucial signaling hub that integrates extracellular cues to coordinate development and maintain homeostasis, and its formation (ciliogenesis) is strictly regulated. Given the complex and spatially distinct organization of the actin filament cytoskeleton, the precise contributions of branched F-actin to ciliogenesis remain poorly understood. Here, we show that centrosomal branched actin filaments (c-BFA) facilitate ciliogenesis by directing the ARP2/3 nucleation‑promoting factor VCA to the centrosome. We identify LRCH2 as an actin‑binding protein that stabilizes c-BFA, thereby promoting ciliary vesicle docking at the mother centriole and enhancing ciliogenesis. In contrast, CCDC22, a known early endosome‑localized protein, regulates endosomal vesicle sorting by stabilizing non‑centrosomal branched actin filaments (nc-BFA), which consequently suppresses ciliary vesicle recruitment to the centrosome and inhibits ciliogenesis. Collectively, LRCH2 and CCDC22 antagonistically regulate branched actin networks at the centrosome and in the cytoplasm, thereby maintaining ciliogenesis homeostasis. Conditional knockout of Ccdc22 in mouse cerebellar granule cell precursors results in an increased proportion of ciliated cells and impairs balance function. Our findings highlight the distinct and opposing roles of c-BFA and nc-BFA in primary ciliogenesis.","manuscriptTitle":"Opposing roles of centrosomal and non-centrosomal branched F-actin networks in primary ciliogenesis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-29 05:07:50","doi":"10.21203/rs.3.rs-9467244/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":"b218a01e-0613-4c39-9c64-7fc9295d30ae","owner":[],"postedDate":"April 29th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":67176520,"name":"Biological sciences/Cell biology/Cytoskeleton/Cilia"},{"id":67176521,"name":"Biological sciences/Cell biology/Cytoskeleton/Actin"}],"tags":[],"updatedAt":"2026-04-29T05:07:50+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-29 05:07:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9467244","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9467244","identity":"rs-9467244","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}