Human O-GlcNAcase catalytic-stalk dimer anchors flexible histone binding domains

Human O-GlcNAcase catalytic-stalk dimer anchors flexible histone binding domains | 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 Human O-GlcNAcase catalytic-stalk dimer anchors flexible histone binding domains John Hanover, Sarah Nyenhuis, Agata Steenackers, Jenny Hinshaw This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6197257/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Dec, 2025 Read the published version in Communications Chemistry → Version 1 posted You are reading this latest preprint version Abstract O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) perform essential functions in signaling, epigenetics, and transcription. Although thousands of proteins are specifically O-GlcNAc modified, the molecular features recognized by the enzymes of O-GlcNAc cycling remain poorly defined. Here we solved the structure of the long isoform of human OGA by cryo-electron microscopy providing a physiologically relevant platform to study the enzyme. The 3.63 Å structure of the dimeric catalytic-stalk dimer differs notably from existing crystal structures. Intriguingly, a low-resolution structure of the OGA-L exhibit densities corresponding to the C-terminal pseudo-HAT domains suggesting substantial flexion of these domains relative to the catalytic-stalk dimer. To explore the role of these domains we found that OGA-L binds to a small subset of the 384 modified histone tails on a commercial array. High specificity binding was observed with modified histone H3K36 peptides, and H4 acetylated peptides. Based on these findings, we propose OGA-L is poised to bind two modified histone tails of nucleosomes in open chromatin but excluded from repressive chromatin. Increased local concentration and activation of OGA-L coupled with its intrinsic conformational flexibility facilitates the removal of O-GlcNAc from target proteins in proximity such as the intrinsically disordered CTD domain of RNA Polymerase II. This model is consistent with OGA-L acting as a ‘reader’ of histone modifications linked to development, transcriptional activation, transposon silencing, and DNA damage repair. Biological sciences/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Biochemistry/Glycobiology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Video1.mp4 Video 1 Cite Share Download PDF Status: Published Journal Publication published 09 Dec, 2025 Read the published version in Communications Chemistry → 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-6197257","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":436565263,"identity":"4d3c5a57-3eaf-46f5-9385-e98df35eaa64","order_by":0,"name":"John Hanover","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCElEQVRIiWNgGAWjYBACxgYeBmaGigOMDewNDAwJbMwGQLE2IrScAWrhOUCkFgYGoBbGNqAWiQQgB6yFgQ2vBub23mOPC+fdkZ3vc4DtwYMya2MG6ea2Bwy/bBIbcDms51y68cxtz4w3Hm9gN0g4l27GIHOw3YCxLw23lhk5ZtK82w4nbuw5wCaR2HbYhgFISjD2HDbG6X2wljlALTMSSNLScDhxvgREixlYC8OPw3I4tfScMZPmOfbMeAPPwTYJoF+MQRolEhvScGoxbO8BaqkBhlh78zHJH2XWhv0S6c8kPvyx4cGppQHKMABGDZgBjpREPLEpD2c0oIj/wa1lFIyCUTAKRhwAAIbAWsogyaecAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3154-9878","institution":"Laboratory of Cell and Molecular Biology, NIDDK, National Institutes of Health","correspondingAuthor":true,"prefix":"","firstName":"John","middleName":"","lastName":"Hanover","suffix":""},{"id":436565264,"identity":"7442a33a-7168-4612-b3dc-92f6a8ec81ab","order_by":1,"name":"Sarah Nyenhuis","email":"","orcid":"https://orcid.org/0000-0002-3139-4423","institution":"National Institutes of Health","correspondingAuthor":false,"prefix":"","firstName":"Sarah","middleName":"","lastName":"Nyenhuis","suffix":""},{"id":436565265,"identity":"5f19de3c-d0c3-4659-a32a-3a901b7a8104","order_by":2,"name":"Agata Steenackers","email":"","orcid":"","institution":"Translational Genomics Center","correspondingAuthor":false,"prefix":"","firstName":"Agata","middleName":"","lastName":"Steenackers","suffix":""},{"id":436565266,"identity":"5f9b1620-c6aa-4a96-9733-9608228a9506","order_by":3,"name":"Jenny Hinshaw","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jenny","middleName":"","lastName":"Hinshaw","suffix":""}],"badges":[],"createdAt":"2025-03-10 16:45:35","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6197257/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6197257/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s42004-025-01813-7","type":"published","date":"2025-12-09T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":99677098,"identity":"8387c676-9d14-4c3f-9142-e2aa3289dbae","added_by":"auto","created_at":"2026-01-07 08:13:59","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":9223611,"visible":true,"origin":"","legend":"Article File","description":"","filename":"Revisedversion3102025withfigures.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6197257/v1_covered_9e46dc7c-fec9-485f-848d-97b0e91f6a9e.pdf"},{"id":79664010,"identity":"385d9b14-82e0-41f8-be00-d5985ffdb4c1","added_by":"auto","created_at":"2025-04-01 09:58:59","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":15750254,"visible":true,"origin":"","legend":"Video 1","description":"","filename":"Video1.mp4","url":"https://assets-eu.researchsquare.com/files/rs-6197257/v1/942459336bb61936566453d5.mp4"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Human O-GlcNAcase catalytic-stalk dimer anchors flexible histone binding domains","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"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-6197257/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6197257/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) perform essential functions in signaling, epigenetics, and transcription. 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