An adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress

An adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress | 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 An adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress Zhi-Hui Zhu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9289811/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 The nucleolus is a membrane-less organelle formed through molecular mechanism of liquid-liquid phase separation of its components from the surrounding nucleoplasm. Nucleolar stress in cancer or tumor cells had been well studied, but the dynamic reorganization process and its molecular and cellular mechanisms involved in phase transition of nucleoli in mouse embryonic stem cells had not been elucidated. Herein, we generate a nucleoli reorganization model in mESCs via a small chemical CX induced inhibitory rRNA biogenesis. We depicted critical nucleolar components repartition upon stress, companied with phase transition in granular component (GC). We suggest the segregation of dense fibrillary component (DFC) is responsible for more liquid-like phase transition in GC, except for the suspicion of lower cellular salt concentration and/or accumulated rRNA induced by CX, which is verified by GC protein NPM1 phase diagram in vitro. Administration of HEX conducts nucleoli an alternative phase transition under stress. Non-physiological GC protein accumulation and interfered hydrophobic microenvironment by HEX contribute its assembles curing and over-condensation in vitro, leading to solid-like phase transition of GC in HEX-plus CX mESCs. Stem Cell & Developmental Cell Biology Mouse embryonic stem cells nucleolar segregation rRNA biosynthesis CX5461 1’6-hexianediol LLPS granular components NPM1 Full Text Additional Declarations The authors declare no competing interests. Supplementary Files 20260401SupplementalMaterialsby.docx An adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress 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. 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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-9289811","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":615838067,"identity":"643bebc4-b189-4cb4-9e9b-bd9b9ac2e252","order_by":0,"name":"Zhi-Hui Zhu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7klEQVRIiWNgGAWjYDACCQYGZgaDAwxs8ocPPvhgwCADFDMgUosEW7LhDAMGHiK1MBwAMnjMpIHqCWuRn91j/Lmg4E5in3SDsbFNgR0Pv0TyBoYfFdtwamGcc8bAeIbBs8Q2mQOJj3MMknkkZ6QVMPacuY1TC7MESJnB4cQ2hoTDxjkGB3gMbuQYMDO24dbCBtRyGKIlsU3aghgtPBI5hs1gLRLJbNIMxGiRkEgrZgZqMW7jOcZs2APyS8+zgoP4/CI/I3nzZ54/h2Xnt/d/fPDjj50cP3vyxgc/KnBrwQ4OkKh+FIyCUTAKRgEaAACrClHAhR0vHwAAAABJRU5ErkJggg==","orcid":"","institution":"college of animal sciences, Zhejiang University","correspondingAuthor":true,"prefix":"","firstName":"Zhi-Hui","middleName":"","lastName":"Zhu","suffix":""}],"badges":[],"createdAt":"2026-04-01 09:37:39","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-9289811/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9289811/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106094183,"identity":"5d3aaddc-127e-4a3e-aeec-2f25b65a340a","added_by":"auto","created_at":"2026-04-03 11:41:30","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2011843,"visible":true,"origin":"","legend":"","description":"","filename":"202600401MSrevision1stby.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9289811/v1_covered_8076e099-b36e-464f-bc4d-d06cc864ee8c.pdf"},{"id":106022565,"identity":"d9f10a04-7e88-4a6d-a029-8b19c1e342cf","added_by":"auto","created_at":"2026-04-02 14:02:31","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":483251,"visible":true,"origin":"","legend":"\u003cp\u003eAn adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress\u003c/p\u003e","description":"","filename":"20260401SupplementalMaterialsby.docx","url":"https://assets-eu.researchsquare.com/files/rs-9289811/v1/2ce78b399b65bb745535181e.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eAn adaptive phase transition in GC is responsible for nucleoli reorganization upon nucleolar stress\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"college of animal sciences, Zhejiang University","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":"Mouse embryonic stem cells, nucleolar segregation, rRNA biosynthesis, CX5461, 1’6-hexianediol, LLPS granular components, NPM1","lastPublishedDoi":"10.21203/rs.3.rs-9289811/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9289811/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe nucleolus is a membrane-less organelle formed through molecular mechanism of liquid-liquid phase separation of its components from the surrounding nucleoplasm. 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