A direct interaction of JAM-C with the tight junction scaffold protein ZO-2

A direct interaction of JAM-C with the tight junction scaffold protein ZO-2 | 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 A direct interaction of JAM-C with the tight junction scaffold protein ZO-2 Annika Schulte, Mariel F. Schwietzer, Frauke Brinkmann, Valentin Teuber, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9013617/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Tight junctions are sites of cell-cell contacts at the apical region of epithelial intercellular junctions that are involved in barrier formation, cellular signaling and cell-cell adhesion. Tight Junctions are formed by integral membrane proteins associated with cytoplasmic scaffolding and adapter proteins through which they are linked to the underlying actomyosin and microtubule cytoskeletons. Here, we have addressed the interaction of the Junctional Adhesion Molecule (JAM)-C with the zonula adherens (ZO) protein ZO-2. Using a combination of cell-based recruitment assays and biochemical in vitro experiments, we find that JAM-C and ZO-2 directly interact in a PDZ domain-dependent manner. Notably, the interaction requires PDZ domain 3 as well as the SH3 domain of ZO-2 indicating that ZO-2 forms a functional supramodule to interact with JAM-C. We also find that JAM-C recruits cingulin to cell-cell contacts, and that JAM-C localization at tight junctions is significantly upregulated in the absence of JAM-A. Our findings have implications on importants aspects of tight junction biology including mechanosensing and liquid-liquid phase separation. Biological sciences/Biochemistry Biological sciences/Biophysics Biological sciences/Cell biology JAM-A JAM-C PDZ domain tight junction ZO-2 Full Text Additional Declarations No competing interests reported. Supplementary Files JAMCZO2SupplMat1.docx JCZO2SourceFileKEfinal.xlsx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 30 Mar, 2026 Reviews received at journal 28 Mar, 2026 Reviews received at journal 18 Mar, 2026 Reviewers agreed at journal 17 Mar, 2026 Reviewers agreed at journal 17 Mar, 2026 Reviewers invited by journal 17 Mar, 2026 Editor assigned by journal 16 Mar, 2026 Editor invited by journal 16 Mar, 2026 Submission checks completed at journal 12 Mar, 2026 First submitted to journal 12 Mar, 2026 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-9013617","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":607893908,"identity":"a6ce84da-92ed-4933-bc8d-abc56498b240","order_by":0,"name":"Annika Schulte","email":"","orcid":"","institution":"ZMBE; University Münster","correspondingAuthor":false,"prefix":"","firstName":"Annika","middleName":"","lastName":"Schulte","suffix":""},{"id":607893910,"identity":"fcdefa06-fed7-465d-adfc-c8cc53a675c9","order_by":1,"name":"Mariel F. 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