Analysis of substrate recognition by Mycobacterium protein-mannosyl transferase reveals WW-domain-like recognition in bacteria

Analysis of substrate recognition by Mycobacterium protein-mannosyl transferase reveals WW-domain-like recognition in bacteria | 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 Analysis of substrate recognition by Mycobacterium protein-mannosyl transferase reveals WW-domain-like recognition in bacteria Michel RIVIERE, Nicolas Géraud, Chloé Rivière, Camille Falcou, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5836107/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Aug, 2025 Read the published version in Communications Biology → Version 1 posted You are reading this latest preprint version Abstract We have previously demonstrated that protein-O-mannosylation (POM), a widespread post-translational glycosyl modification of proteins, is a key virulence factor of Mycobacterium tuberculosis (Mtb), the world’s deadliest infectious agent. Here, we report a detailed analysis of the structure-function relationship of MtPMT, the enzyme that catalyzes POM in Mtb. Using mutagenesis and in cellulo monitoring of POM activity, we demonstrate that, despite notable structural differences, MtPMT shares functional homologies with yeasts’ PMTs in the mechanism of the sugar transfer from lipidic donors. Furthermore, we provide evidence that the selectivity for proline-rich target glycosylation sites that differentiates MtPMT from its eukaryotic homologues, relies on a WW-like domain, which preferentially interacts with proline-rich acceptor substrate analogues. This first identification of a functional WW-like domain in a prokaryotic protein raises questions about its potential evolutionary linkage with eukaryotic WW modules and provides new insights into PMT’s acceptor-substrate recognition mechanism paving the way for the development selective inhibitors of MtPMT with potential therapeutic application against tuberculosis. Biological sciences/Molecular biology/Post-translational modifications/Glycosylation Biological sciences/Microbiology/Pathogens Full Text Additional Declarations There is NO Competing Interest. Supplementary Files TableS2Mtbmannoproteinsforfig4a.xlsx TableS5PhylogenySequencesforfig7.xlsx GeraudetalSupMaterial.pdf Cite Share Download PDF Status: Published Journal Publication published 07 Aug, 2025 Read the published version in Communications Biology → 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. 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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-5836107","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":406235337,"identity":"66e1d61c-c835-4182-b684-d836959ac6a4","order_by":0,"name":"Michel RIVIERE","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIie3RMQrCMBSA4SeFuATnlAqeQHguiiB6FUXQRdCxg0NDoV08gINHcUgpOBVdBZeK0Fk3BRFTK4qDzSqYf0gTyEdICqDT/WIGEIAJyLHgyGWLApMfW0nwRQYZifLPycizEJSk6htJfEQYl1ifH2x7U25YfgzB8juph6RRmyM0PdZza1G0o81FhCCSPEKJRRGQ0MAzubejuB2BcRQKcs2If+G3tSTDGISKQEqK3CtwR0jSRQUhdXOGLCWu6az6j7uIXLIJE3a2W1hxi/uTM2135Ivt4zzyjL2n6T9Sg49QvUWn0+n+rDv120ynHtUS3gAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-3788-0588","institution":"CNRS - 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