Zinc-redox crosstalk regulates proteostasis in the endoplasmic reticulum

Zinc-redox crosstalk regulates proteostasis in the endoplasmic reticulum | 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 Zinc-redox crosstalk regulates proteostasis in the endoplasmic reticulum Kenji Inaba, Yuta Amagai, Chihiro Arai, Wakana Yamamoto, Satoshi Watanabe, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7251469/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Zinc homeostasis is crucial for various biological processes, including gene regulation, signal transduction, and proteostasis. ZIP7 is a membrane transporter that exports zinc ions (Zn 2+ ) from the lumen of the endoplasmic reticulum (ER) to the cytosol, and its dysfunction causes ER stress, although the underlying mechanism remains unclear. Here, we show that ZIP7 inhibition increases the labile Zn 2+ concentration in the ER to micromolar levels, approximately 10 6 times higher than its steady-state level. Such abnormally high Zn 2+ concentrations disrupt the function and trafficking of the Zn 2+ -dependent chaperone ERp44 at the ER-Golgi interface. In vitro assays using recombinant proteins demonstrated that Zn 2+ inhibits the Ero1α-PDI oxidative system, and that ERp44 enhances this inhibitory effect. Consequently, the ER redox environment becomes more reducing, severely impairing the oxidative folding of key membrane receptors such as Notch1 and EGFR. These findings reveal the essential role of zinc homeostasis in redox-dependent proteostasis within the ER. Biological sciences/Cell biology/Organelles/Endoplasmic reticulum Biological sciences/Biochemistry/Protein folding/Endoplasmic reticulum EGF receptor ERp44 Notch receptor Oxidative protein folding Proteostasis Redox Zinc ZIP7 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files VideoS1.mp4 Supplementary movie 1 VideoS2.mp4 Supplementary movie 2 VideoS3.mp4 Supplementary movie 3 VideoS4.mp4 Supplementary movie 4 VideoS5.mp4 Supplementary movie 5 AmagaiEtAlZIP7redoxSupplement.docx Supplementary information Cite Share Download PDF Status: Under Review 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-7251469","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":498701674,"identity":"91de66f7-c516-4034-8fc7-aded107945ab","order_by":0,"name":"Kenji 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ZIP7","lastPublishedDoi":"10.21203/rs.3.rs-7251469/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7251469/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Zinc homeostasis is crucial for various biological processes, including gene regulation, signal transduction, and proteostasis. ZIP7 is a membrane transporter that exports zinc ions (Zn\u003csup\u003e2+\u003c/sup\u003e) from the lumen of the endoplasmic reticulum (ER) to the cytosol, and its dysfunction causes ER stress, although the underlying mechanism remains unclear. Here, we show that ZIP7 inhibition increases the labile Zn\u003csup\u003e2+\u003c/sup\u003e concentration in the ER to micromolar levels, approximately 10\u003csup\u003e6\u003c/sup\u003e times higher than its steady-state level. Such abnormally high Zn\u003csup\u003e2+\u003c/sup\u003e concentrations disrupt the function and trafficking of the Zn\u003csup\u003e2+\u003c/sup\u003e-dependent chaperone ERp44 at the ER-Golgi interface. In vitro assays using recombinant proteins demonstrated that Zn\u003csup\u003e2+\u003c/sup\u003e inhibits the Ero1α-PDI oxidative system, and that ERp44 enhances this inhibitory effect. Consequently, the ER redox environment becomes more reducing, severely impairing the oxidative folding of key membrane receptors such as Notch1 and EGFR. 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