Ran Modulates Allosteric Crosstalk Between Importin β Surfaces

Ran Modulates Allosteric Crosstalk Between Importin β Surfaces | 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 Ran Modulates Allosteric Crosstalk Between Importin β Surfaces Gino Cingolani, Ying-Hui Ko, Fenglin Li, Stephanie Suinn, Junwei Li, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6449265/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 11 Dec, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract A cellular gradient of the GTPase Ran orchestrates the movement of import and export complexes through the Nuclear Pore Complex (NPC). Ran-GTP modulates two essential activities of importin β for nuclear import. On one hand, it reduces the avidity of importin β for phenylalanine-glycine-rich nucleoporins (FG-nups), facilitating the passage of import complexes through the permeability barrier; on the other hand, it disassembles import complexes, releasing the import cargo into the nucleus. The precise mechanisms by which Ran-GTP modulates importin β activities remain hypothetical. Leveraging cryogenic electron microscopy (cryo-EM) single particle analysis, in this paper, we describe four distinct conformational states of importin β in complex with binding effectors encountered during an import reaction, specifically IBB-cargos, FG-repeats, Ran-GTP, and Ran-GTP:RanBP1. Comparing these four states enables us to decipher the conformational landscape of importin β without interference from crystallization agents and lattice forces. By correlating structural data with biochemical activities, we find that Ran-GTP constrains the solenoid structure of importin β, closing four high-affinity FG-binding pockets and displacing import cargos through allosteric crosstalk between the concave and convex surfaces. We propose that this allosteric mechanism is relevant to other β-karyopherins involved in nuclear import. Biological sciences/Biochemistry/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Cell biology/Nuclear transport/Protein transport/Protein translocation Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files ImpBSUP4122025update.docx RAN MODULATES ALLOSTERIC CROSSTALK BETWEEN IMPORTIN β SURFACES Cite Share Download PDF Status: Published Journal Publication published 11 Dec, 2025 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6449265","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":447571195,"identity":"9767031c-31a8-4b6c-ac29-ca51365ed2d4","order_by":0,"name":"Gino 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