CHAMP1 premature termination codon mutations found in individuals with intellectual disability cause a homologous recombination defect through haploinsufficiency

CHAMP1 premature termination codon mutations found in individuals with intellectual disability cause a homologous recombination defect through haploinsufficiency | 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 CHAMP1 premature termination codon mutations found in individuals with intellectual disability cause a homologous recombination defect through haploinsufficiency Yujiro Yoshizaki, Yunosuke Ouchi, Dicky Kurniawan, Eisuke Yumoto, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4346414/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Dec, 2024 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract CHAMP1 (chromosome alignment-maintaining phosphoprotein 1) plays a role in the repair of DNA double-strand breaks (DSBs) by homologous recombination (HR). The CHAMP1 gene is one of the genes mutated in individuals with intellectual disability. The majority of the mutations are premature termination codon (PTC) mutations, while missense mutations have also been reported. How these mutations affect the functions of CHAMP1 has not been clarified yet. Here we investigated the effects of the CHAMP1 mutations on HR. In B lymphocytes and fibroblasts derived from individuals with CHAMP1 PTC mutations, truncated CHAMP1 proteins of the expected sizes were detected. When DSBs were induced in fibroblasts with PTC mutations, a defect in HR was detected. U2OS cells expressing the CHAMP1 mutants did not show an HR defect in the presence of endogenous wild-type (WT) CHAMP1, whereas they were unable to restore HR activity when WT CHAMP1 was depleted, suggesting that the PTC mutations are loss-of-function mutations. On the other hand, the CHAMP1 mutants with missense mutations restored HR activity when WT CHAMP1 was depleted. In DLD-1 cells, heterozygous depletion of CHAMP1 resulted in an HR defect, indicating haploinsufficiency. These results suggest that CHAMP1 PTC mutations cause an HR defect through a haploinsufficient mechanism, while CHAMP1 missense mutations do not affect the HR function of CHAMP1. CHAMP1 DNA double strand breaks homologous recombination intellectual disability haploinsufficiency camptothecin Full Text Additional Declarations No competing interests reported. Supplementary Files 20240507SupplementaryInformation.pdf Cite Share Download PDF Status: Published Journal Publication published 30 Dec, 2024 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 10 Jun, 2024 Reviews received at journal 07 Jun, 2024 Reviewers agreed at journal 27 May, 2024 Reviews received at journal 27 May, 2024 Reviewers agreed at journal 17 May, 2024 Reviewers invited by journal 08 May, 2024 Editor assigned by journal 08 May, 2024 Editor invited by journal 08 May, 2024 Submission checks completed at journal 08 May, 2024 First submitted to journal 30 Apr, 2024 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-4346414","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":301894786,"identity":"f749b992-9e44-435e-b573-13fb2aa53001","order_by":0,"name":"Yujiro Yoshizaki","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Yujiro","middleName":"","lastName":"Yoshizaki","suffix":""},{"id":301894787,"identity":"567dc5f1-af57-4ce1-a0bc-7437fa36e9bb","order_by":1,"name":"Yunosuke Ouchi","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Yunosuke","middleName":"","lastName":"Ouchi","suffix":""},{"id":301894788,"identity":"21b8286a-97be-4444-bda7-88b8065acbb8","order_by":2,"name":"Dicky Kurniawan","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Dicky","middleName":"","lastName":"Kurniawan","suffix":""},{"id":301894789,"identity":"c94196f4-752a-489a-96e9-a6121e55c672","order_by":3,"name":"Eisuke Yumoto","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Eisuke","middleName":"","lastName":"Yumoto","suffix":""},{"id":301894790,"identity":"3d18beeb-5a7c-46e1-9db7-ef8289d3f4f6","order_by":4,"name":"Yuki Yoneyama","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Yuki","middleName":"","lastName":"Yoneyama","suffix":""},{"id":301894791,"identity":"8c2d1ccd-c0f7-453c-aa7b-f389edd0487a","order_by":5,"name":"Faiza Ramadhani Rizqullah","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Faiza","middleName":"Ramadhani","lastName":"Rizqullah","suffix":""},{"id":301894792,"identity":"1d118fff-747f-4e1a-9653-9035c93f6c6b","order_by":6,"name":"Hiyori Sato","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Hiyori","middleName":"","lastName":"Sato","suffix":""},{"id":301894793,"identity":"59212ad9-ec8c-4260-aa0b-271450e2a4f2","order_by":7,"name":"Mirjam Hanako Sarholz","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Mirjam","middleName":"Hanako","lastName":"Sarholz","suffix":""},{"id":301894794,"identity":"f73a6ed1-80d8-4865-a2f3-4fa2aa21e564","order_by":8,"name":"Toyoaki Natsume","email":"","orcid":"","institution":"National Institute of Genetics","correspondingAuthor":false,"prefix":"","firstName":"Toyoaki","middleName":"","lastName":"Natsume","suffix":""},{"id":301894795,"identity":"bc3fcc9d-8edc-4f5a-8101-b51afc60330b","order_by":9,"name":"Masato T. Kanemaki","email":"","orcid":"","institution":"National Institute of Genetics","correspondingAuthor":false,"prefix":"","firstName":"Masato","middleName":"T.","lastName":"Kanemaki","suffix":""},{"id":301894796,"identity":"74767f37-ad97-4471-a39a-810a238c3ff3","order_by":10,"name":"Masanori Ikeda","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Masanori","middleName":"","lastName":"Ikeda","suffix":""},{"id":301894797,"identity":"d756dbdd-a44f-4c13-a511-89b9a9d4da5b","order_by":11,"name":"Ayako Ui","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Ayako","middleName":"","lastName":"Ui","suffix":""},{"id":301894798,"identity":"51baa352-67fc-4df2-b192-1e032a708741","order_by":12,"name":"Kenji Iemura","email":"","orcid":"","institution":"Tohoku University","correspondingAuthor":false,"prefix":"","firstName":"Kenji","middleName":"","lastName":"Iemura","suffix":""},{"id":301894799,"identity":"03c1dbab-8b20-408c-9cc1-b0f16262c70f","order_by":13,"name":"Kozo Tanaka","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABFklEQVRIie2QsWrDMBBAzwjSRUXrGUPzCzJZg79FQuAupRS6BAJNoOAs/oAWTPsLzR+4COxFjtdCFndPIWOnUiXEJYNdyFaKHgiO497d6QAcjj8I/kR09ybjXYocUvkvimgVE+OJynmij5Vu/MXivfE+o2uW6gY2g/qOBfd6RCEagld1jglorjgIdYtVwr2MrtHPCqUoqHAOK9GlXKAoEASR8xo4obhG/nY10hSILTe8W5GJVWbyuT7bEspXrTLrVQJUA6to+VKldorI94pdTPcqfloQLuJSLo25ec1y5T8+xCrMeBkmPX/B0l5sO57KJ3O5bD6+IsZQadxMpkOG3Rfb0zY7qrArDdD0Gn2w9GTF4XA4/iXfrEtbtR2nyV8AAAAASUVORK5CYII=","orcid":"","institution":"Tohoku University","correspondingAuthor":true,"prefix":"","firstName":"Kozo","middleName":"","lastName":"Tanaka","suffix":""}],"badges":[],"createdAt":"2024-04-30 05:31:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4346414/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4346414/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-024-83435-y","type":"published","date":"2024-12-30T15:56:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":73093486,"identity":"1cce35a6-f362-470e-a938-d5d50367c59a","added_by":"auto","created_at":"2025-01-06 16:20:49","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2763884,"visible":true,"origin":"","legend":"","description":"","filename":"240507ManuscriptYoshizakietal.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4346414/v1_covered_ad44aa3f-db4d-45ba-ae10-45910d52d256.pdf"},{"id":56481222,"identity":"da84f94f-0253-4cc0-8913-9ce673333319","added_by":"auto","created_at":"2024-05-14 18:31:28","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":9958293,"visible":true,"origin":"","legend":"","description":"","filename":"20240507SupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4346414/v1/71dc1b06ee148d84d6d9e7cc.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"CHAMP1 premature termination codon mutations found in individuals with intellectual disability cause a homologous recombination defect through haploinsufficiency","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"CHAMP1, DNA double strand breaks, homologous recombination, intellectual disability, haploinsufficiency, camptothecin","lastPublishedDoi":"10.21203/rs.3.rs-4346414/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4346414/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"CHAMP1 (chromosome alignment-maintaining phosphoprotein 1) plays a role in the repair of DNA double-strand breaks (DSBs) by homologous recombination (HR). The CHAMP1 gene is one of the genes mutated in individuals with intellectual disability. The majority of the mutations are premature termination codon (PTC) mutations, while missense mutations have also been reported. How these mutations affect the functions of CHAMP1 has not been clarified yet. Here we investigated the effects of the CHAMP1 mutations on HR. In B lymphocytes and fibroblasts derived from individuals with CHAMP1 PTC mutations, truncated CHAMP1 proteins of the expected sizes were detected. When DSBs were induced in fibroblasts with PTC mutations, a defect in HR was detected. U2OS cells expressing the CHAMP1 mutants did not show an HR defect in the presence of endogenous wild-type (WT) CHAMP1, whereas they were unable to restore HR activity when WT CHAMP1 was depleted, suggesting that the PTC mutations are loss-of-function mutations. On the other hand, the CHAMP1 mutants with missense mutations restored HR activity when WT CHAMP1 was depleted. In DLD-1 cells, heterozygous depletion of CHAMP1 resulted in an HR defect, indicating haploinsufficiency. These results suggest that CHAMP1 PTC mutations cause an HR defect through a haploinsufficient mechanism, while CHAMP1 missense mutations do not affect the HR function of CHAMP1.","manuscriptTitle":"CHAMP1 premature termination codon mutations found in individuals with intellectual disability cause a homologous recombination defect through haploinsufficiency","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-14 18:31:23","doi":"10.21203/rs.3.rs-4346414/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-06-10T08:26:47+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-06-07T10:07:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"197385863483441454629523121504367625441","date":"2024-05-27T21:59:11+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-05-27T08:33:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"52511675313122848274027049528676638079","date":"2024-05-17T08:30:22+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-05-08T12:08:42+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-05-08T12:02:40+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-05-08T11:58:34+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-05-08T04:47:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-04-30T05:30:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4e546e36-c375-401b-8411-10ce40681c20","owner":[],"postedDate":"May 14th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-01-06T16:04:54+00:00","versionOfRecord":{"articleIdentity":"rs-4346414","link":"https://doi.org/10.1038/s41598-024-83435-y","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2024-12-30 15:56:56","publishedOnDateReadable":"December 30th, 2024"},"versionCreatedAt":"2024-05-14 18:31:23","video":"","vorDoi":"10.1038/s41598-024-83435-y","vorDoiUrl":"https://doi.org/10.1038/s41598-024-83435-y","workflowStages":[]},"version":"v1","identity":"rs-4346414","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4346414","identity":"rs-4346414","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}