Human ribosome interactions reframe neomycin toxicity

Human ribosome interactions reframe neomycin toxicity | 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 Biological Sciences - Article Human ribosome interactions reframe neomycin toxicity Wenfei Li, Alexey Amunts, Meng Wu, Xuben Hou, Lei Guo, Zhe Zhang, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7177598/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 Aminoglycosides like neomycin are widely used but clinically limited due to toxicity, traditionally attributed to mitoribosome inhibition. Here, we demonstrate that this assumption requires re-evaluation by comparing neomycin’s interactions with human ribosomes in vitro and in cells. While cryo-EM and biochemical assays reveal strong in vitro binding to both mitoribosomes and cytosolic ribosomes, especially at conserved regions such as the helix 44 (h44) decoding center despite sequence divergence, and H69. Cellular analyses show minimal impact on global translation, reduced occupancy on cytosolic ribosomes, and a complete absence of neomycin on mitoribosomes. These discrepancies suggest limited mitochondrial permeability rather than direct mitoribosome inhibition underlies neomycin’s toxicity. Our findings redefine the mechanistic basis of aminoglycosides side effects and call for a reassessment of their cellular targets. Biological sciences/Biochemistry/Structural biology/Electron microscopy/Cryoelectron microscopy Biological sciences/Structural biology/Electron microscopy/Cryoelectron microscopy Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryData.pdf Supplementary figures and tables 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. 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-7177598","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":490545070,"identity":"72dbb617-869c-4d8a-ba1a-db346056c88b","order_by":0,"name":"Wenfei 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