Mechanisms of Tuning Tension at the Centromeric Nucleosome

Mechanisms of Tuning Tension at the Centromeric Nucleosome | 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 Research Article Mechanisms of Tuning Tension at the Centromeric Nucleosome Aryan Kokkanti, Daniel Kolbin, Kerry Bloom This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9478713/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 6 You are reading this latest preprint version Abstract The centromere is chromosomal locus that provides the site of attachment for kinetochore microtubules. In budding yeast, the centromere is defined as a 120-bp segment containing three conserved DNA elements (CDEs) I, II and III. The centromere is wrapped around a histone octamer containing a centromere-specific histone H3 variant (Cse4, CENP-A). We determine that the centromere VIII 118-bp conserved sequence is insufficient for centromere function. The addition of at least 19 bp of DNA flanking centromere DNA element III (CDEIII) of the endogenous sequence was necessary for full segregation function. Using a dicentric chromosome competition assay to mechanically stress two centromeres on a single chromosome, we find an essential role for excess, unbound Cse4/CENP-A. These data suggest that centromere DNA to CENP-A nucleosome binding is multivalent and dynamic with histone dissociation facilitated by the local concentration of unbound Cse4/CENP-A. Competition between bound and unbound Cse4 drives transient cycles of centromere nucleosome assembly and disassembly, which induce DNA length extension and retraction in concert with a dynamically growing and shortening kinetochore-microtubule plus-end. Our studies reveal how histone exchange within the centromere nucleosome may regulate centromere DNA extension at the nanometer scale, coupling it to the commensurate nanoscale dynamics of microtubule plus ends. Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryDataFile.xlsx SupplementaryTableFigs.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 30 Apr, 2026 Reviewers agreed at journal 29 Apr, 2026 Reviewers invited by journal 29 Apr, 2026 Editor assigned by journal 28 Apr, 2026 Submission checks completed at journal 23 Apr, 2026 First submitted to journal 21 Apr, 2026 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-9478713","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":632559065,"identity":"9a01c80d-13ee-4295-bd1d-1a382a9fe029","order_by":0,"name":"Aryan Kokkanti","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Aryan","middleName":"","lastName":"Kokkanti","suffix":""},{"id":632559066,"identity":"1a0c43db-cb70-4afe-bd31-d5b4d045e989","order_by":1,"name":"Daniel Kolbin","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtUlEQVRIiWNgGAWjYDCCMwyMhxkMGOSgXGaitDCAtBiTqoWBIbGBaC18Zw4fOFxQcCd9w40cwwcMFdYwvbiB5Nm2hMMzDJ7lArUYGzCcSSesxeA8j8FhIMrdcDvHTIKx7TDxWtINwFr+EaPlbA9YSwJESwMRWiTPHAP55bDhzPvPig0SjqUbE9TCdyb54OOCP4flgUG38cGHGmtZglqQAIcBQwIJykGA/QGJGkbBKBgFo2CkAAAUYkaRejHINAAAAABJRU5ErkJggg==","orcid":"","institution":"University of Wyoming","correspondingAuthor":true,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Kolbin","suffix":""},{"id":632559067,"identity":"ec87efb9-8edc-42b9-aa80-82e679c704c3","order_by":2,"name":"Kerry Bloom","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Kerry","middleName":"","lastName":"Bloom","suffix":""}],"badges":[],"createdAt":"2026-04-21 04:53:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9478713/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9478713/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108810035,"identity":"45403714-b846-4b3a-aaef-f18002866210","added_by":"auto","created_at":"2026-05-08 15:57:03","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1624737,"visible":true,"origin":"","legend":"","description":"","filename":"MainTextver14.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9478713/v1_covered_01762ed2-8623-4ce8-b65d-532fce81a24a.pdf"},{"id":108693899,"identity":"c34f10f3-b245-4679-bd9b-c2ea0b62408d","added_by":"auto","created_at":"2026-05-07 11:28:18","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":36670,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryDataFile.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-9478713/v1/238b65a9749af1e9b042dfce.xlsx"},{"id":108806371,"identity":"9f0a6d3b-e9a9-424f-ba07-c7b315bc8e78","added_by":"auto","created_at":"2026-05-08 15:28:24","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":2021542,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryTableFigs.docx","url":"https://assets-eu.researchsquare.com/files/rs-9478713/v1/763be86186ae669a9c0726fe.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Mechanisms of Tuning Tension at the Centromeric Nucleosome","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"chromosome-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"chrs","sideBox":"Learn more about [Chromosome Research](http://link.springer.com/journal/10577)","snPcode":"10577","submissionUrl":"https://submission.nature.com/new-submission/10577/3","title":"Chromosome Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-9478713/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9478713/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe centromere is chromosomal locus that provides the site of attachment for kinetochore microtubules. 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