Unanchored K63-linked polyubiquitin chains: a novel second messenger involved in G protein-coupled receptors early signaling events

Unanchored K63-linked polyubiquitin chains: a novel second messenger involved in G protein-coupled receptors early signaling events | 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 Unanchored K63-linked polyubiquitin chains: a novel second messenger involved in G protein-coupled receptors early signaling events Marc Servant, Priscilla Doyon, Mohamad-Ali El-Mortada This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5891152/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 GPCR rapidly transduced extracellular cues via their ability to activate effector proteins leading to the production of a variety of well-characterized second messenger molecules. We describe here that the E3 ubiquitin ligase tumor necrosis factor receptor associated factor 6 (TRAF6) is required for the rapid and transient production of unanchored lysine (K)63-linked polyubiquitin chains in primary and transformed cells exposed to vasoactive agonists Angiotensin II (Ang II), Lysophosphatidic acid (LPA) and Thrombin (Thr). We further show that unanchored K63-linked polyubiquitin chains accumulate in Transforming Growth Factor β-activated kinase 1 (TAK1) and IkappaB kinase beta (IKKβ) immunocomplexes and are required for the T-loop phosphorylation of TAK1 leading to IKKβ, c- Jun N-terminal kinases (JNK1/2) and p38 mitogen-activated protein kinases (p38) phosphorylation and the induction of transcriptional events. Our results support a novel paradigm in field of GPCR signal transduction, identifying unanchored K63-linked polyubiquitin chains as novel second messenger molecules. Biological sciences/Cell biology/Cell signalling Biological sciences/Biochemistry/Enzyme mechanisms Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SuppFig1.pdf Supplementary Figure 1 Figsup2.pdf Supplementary Figure 2 Figsup3v2.pdf Supplementary Figure 3 Figsup4.pdf Supplementary Figure 4 Figsup5red.pdf Supplementary Figure 5 Figsup6.pdf Supplementary Figure 6 Figsup7.pdf Supplementary Figure 7 Figsup8red.pdf Supplementary Figure 8 Figsup9red.pdf Supplementary Figure 9 Figsup10.pdf Supplementary Figure 10 Figsup11red.pdf Supplementary Figure 11 Figsup12.pdf Supplementary Figure 12 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-5891152","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":413724783,"identity":"3553e9ee-8ac9-4fbd-8fa7-37c2785b2d3c","order_by":0,"name":"Marc 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