Targeted stress granule regulation by engineering a non-catalytic O-GlcNAc transferase

Targeted stress granule regulation by engineering a non-catalytic O-GlcNAc transferase | 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 Targeted stress granule regulation by engineering a non-catalytic O-GlcNAc transferase Yun Ge, Na Wang, Fanjia Hou, Sihui Ma, Silan Liu, Haimei Wei, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6750567/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Jan, 2026 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Stress granules (SGs) are dynamic ribonucleoprotein condensates whose assembly is driven by multivalent interactions and liquid-liquid phase separation (LLPS) of their component proteins and mRNAs. Related to multiple diseases, SGs are an ideal prototype to study cellular condensates. Understanding their regulatory factors and developing modulation methods are critical to therapeutic development. O-GlcNAc transferase (OGT) has been implicated in SG regulation, while its function beyond O-GlcNAcylation remains unknown. Here, we identified OGT to suppress the LLPS of G3BP1, a key SG protein, and thereby SG assembly. Unexpectedly, this suppression is independent of the OGT enzymatic activity. Also, we repurposed OGT into an SG modulator by generating a fusion protein consisting of its N-catalytic domain and poorly understood intervening domain (NI), coupled with induced proximity modules, such as a nanobody. We demonstrate that this inhibitory effect is achieved via targeted protein immobilization, which rigidifies G3BP1 after prolonged stress. This modular, genetically encoded tool recognizes the domain organization of G3BP1, thus is generalizable to another four proteins featuring similar architecture, suppressing condensate formation with mobility reduction. We also applied this strategy for SG perturbations to reveal SG functions on cellular activities. Our work provides a novel strategy for general SG regulation by interfering material properties of critical SG proteins and offers insights into the cryptic non-catalytic function of OGT. Biological sciences/Chemical biology/Glycobiology Biological sciences/Chemical biology/Synthetic biology Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Published Journal Publication published 07 Jan, 2026 Read the published version in Nature Communications → 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-6750567","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":469498405,"identity":"117d9250-02af-458d-b000-069d0ed5366f","order_by":0,"name":"Yun 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