RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity

RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity | 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 RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity Ganesh Nagaraju, Satyaranjan Sahoo, Tarun Nagraj, Debanjali Bhattacharya, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5319894/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Fanconi anemia (FA) is characterized by bone marrow failure, congenital abnormalities, and cancer predisposition. Mutations in RAD51 paralogs have been identified in FA-like disorder and cancers. Although the role of RAD51 paralogs is well established in homologous recombination-mediated DNA repair, little is known about their role during replication stress responses. Here, we report that the RAD51C/XRCC3 (CX3) complex of RAD51 paralogs participates in the FA-pathway of R-loop tolerance mechanism. CX3 complex suppresses R-loops, transcription-replication collisions (TRC), and associated genome instability in physiological and replication stress conditions. Mechanistically, the CX3 complex physically interacts with FANCM and facilitates its recruitment to the R-loop sites to promote its resolution. Notably, cells expressing RAD51C R258H pathological mutant exhibit defective interaction with FANCM and display inefficient R-loop processing. This work identifies a novel role of the CX3 complex in preventing R-loop-induced genome instability by regulating FANCM-mediated R-loop resolution. Biological sciences/Molecular biology/DNA damage and repair Biological sciences/Cell biology/Mechanisms of disease Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Posted 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-5319894","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":372271289,"identity":"0324e95f-5dff-4955-bdde-1122796b2dda","order_by":0,"name":"Ganesh Nagaraju","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAw0lEQVRIiWNgGAWjYPCCA3JszFAmG7FajEnXkthAtIvM+Rc/e/Dhz530Pnb2hx8YauwY+KQJ6Lac8czccGbbs9w2Zh5jCYZjyQxsMgfwazG4ccBMmrfhMEgL0BdsBxjYJBIIaTn+TfrPn8PpbMzszxgY/hGj5XyPmTQD2+EEYIiZMTC2EWULT5lkb9thQ7BfEvuSeYiw5fg2iR9/DsvL9x9/+OHDNzs5+RkEtDCgmAlk8xBQDwT8BwirGQWjYBSMghEOADtgPM8qtl/tAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3386-7095","institution":"Indian Institute of Science Bangalore","correspondingAuthor":true,"prefix":"","firstName":"Ganesh","middleName":"","lastName":"Nagaraju","suffix":""},{"id":372271290,"identity":"cc4d36a9-479d-4bd6-bb72-02a362d04305","order_by":1,"name":"Satyaranjan Sahoo","email":"","orcid":"https://orcid.org/0000-0002-1868-758X","institution":"Indian Institute of Science Bangalore","correspondingAuthor":false,"prefix":"","firstName":"Satyaranjan","middleName":"","lastName":"Sahoo","suffix":""},{"id":372271291,"identity":"d9d25dd6-fbae-4a4c-9b67-04dbf7f01e53","order_by":2,"name":"Tarun Nagraj","email":"","orcid":"","institution":"Indian Institute of Science Bangalore","correspondingAuthor":false,"prefix":"","firstName":"Tarun","middleName":"","lastName":"Nagraj","suffix":""},{"id":372271292,"identity":"ff89d782-a5b3-40ad-8458-94c85701286d","order_by":3,"name":"Debanjali Bhattacharya","email":"","orcid":"","institution":"Indian Institute of Science Bangalore","correspondingAuthor":false,"prefix":"","firstName":"Debanjali","middleName":"","lastName":"Bhattacharya","suffix":""},{"id":372271293,"identity":"5364003b-0149-450c-8d22-e4669308e785","order_by":4,"name":"Kumar Somyajit","email":"","orcid":"https://orcid.org/0000-0003-2496-7155","institution":"University of Southern Denmark","correspondingAuthor":false,"prefix":"","firstName":"Kumar","middleName":"","lastName":"Somyajit","suffix":""}],"badges":[],"createdAt":"2024-10-23 14:50:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5319894/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5319894/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":70421050,"identity":"8d1c8d3b-6f8c-4161-b2cd-ef4e4fddfc14","added_by":"auto","created_at":"2024-12-03 04:22:53","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7388000,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptMainFigSupplFig.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5319894/v1_covered_fec09c7b-14f7-4817-9ffb-63f230bec82d.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-5319894/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5319894/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Fanconi anemia (FA) is characterized by bone marrow failure, congenital abnormalities, and cancer predisposition. Mutations in RAD51 paralogs have been identified in FA-like disorder and cancers. Although the role of RAD51 paralogs is well established in homologous recombination-mediated DNA repair, little is known about their role during replication stress responses. Here, we report that the RAD51C/XRCC3 (CX3) complex of RAD51 paralogs participates in the FA-pathway of R-loop tolerance mechanism. CX3 complex suppresses R-loops, transcription-replication collisions (TRC), and associated genome instability in physiological and replication stress conditions. Mechanistically, the CX3 complex physically interacts with FANCM and facilitates its recruitment to the R-loop sites to promote its resolution. Notably, cells expressing RAD51C R258H pathological mutant exhibit defective interaction with FANCM and display inefficient R-loop processing. This work identifies a novel role of the CX3 complex in preventing R-loop-induced genome instability by regulating FANCM-mediated R-loop resolution.","manuscriptTitle":"RAD51C-XRCC3 complex regulates FANCM-mediated R-loop resolution to safeguard genome integrity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-03 04:14:38","doi":"10.21203/rs.3.rs-5319894/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6a97ac30-48b7-4e5e-8b0d-90d24f05df61","owner":[],"postedDate":"December 3rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":39621905,"name":"Biological sciences/Molecular biology/DNA damage and repair"},{"id":39621906,"name":"Biological sciences/Cell biology/Mechanisms of disease"}],"tags":[],"updatedAt":"2024-12-03T04:14:38+00:00","versionOfRecord":[],"versionCreatedAt":"2024-12-03 04:14:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5319894","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5319894","identity":"rs-5319894","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}