Phosphorylation-mediated conformational switch regulates human SLFN11

Phosphorylation-mediated conformational switch regulates human SLFN11 | 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 Phosphorylation-mediated conformational switch regulates human SLFN11 Katja Lammens, Michael Kugler, Felix Metzner, Karl-Peter Hopfner This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4164465/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 Dec, 2024 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Human Schlafen 11 (SLFN11) serves as a predictive biomarker in cancer as it sensitizes cells to DNA damaging agents by irreversibly blocking stalled replication forks. SLFN11 further acts as an interferon-inducible antiviral restriction factor that targets translation in a codon-usage-dependent manner. However, the regulation of the SLFN11 functions and enzymatic activities remains enigmatic. Here, we present a structural and biochemical explanation for the regulation of SLFN11 by phosphorylation. A cryo-electron microscopy (cryo-EM) structure of the SLFN11 phosphomimetic mutant S753D revealed that it adopts a monomeric conformation, allowing ATP binding, yet loses its ability to bind single-stranded DNA (ssDNA). Cryo-EM structures of SLFN11 bound to tRNA-Leu and tRNA-Met give insights into tRNA binding and cleavage, as well as its regulation by phosphorylation at S219 and T230. Thus, the phosphorylation site S753 serves as a conformational switch, regulating SLFN11 dimerization, as well as ATP and ssDNA binding, while S219 and T230 regulate tRNA recognition and nuclease activity. Biological sciences/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Cancer/Tumour biomarkers Biological sciences/Immunology/Infectious diseases/HIV infections Biological sciences/Immunology/Innate immunity/Pattern recognition receptors Biological sciences/Biochemistry/Enzyme mechanisms Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SISLFN11S753D.pdf ReportingSummary.pdf Reporting Summary Cite Share Download PDF Status: Published Journal Publication published 03 Dec, 2024 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. 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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-4164465","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":295554513,"identity":"ae63e422-70b2-4d77-8167-dabb38c90b2f","order_by":0,"name":"Katja 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SLFN11 further acts as an interferon-inducible antiviral restriction factor that targets translation in a codon-usage-dependent manner. However, the regulation of the SLFN11 functions and enzymatic activities remains enigmatic. Here, we present a structural and biochemical explanation for the regulation of SLFN11 by phosphorylation. A cryo-electron microscopy (cryo-EM) structure of the SLFN11 phosphomimetic mutant S753D revealed that it adopts a monomeric conformation, allowing ATP binding, yet loses its ability to bind single-stranded DNA (ssDNA). Cryo-EM structures of SLFN11 bound to tRNA-Leu and tRNA-Met give insights into tRNA binding and cleavage, as well as its regulation by phosphorylation at S219 and T230. 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