The birth, death, and evolutionary compensation of uORFs in Drosophila

The birth, death, and evolutionary compensation of uORFs in Drosophila | 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 The birth, death, and evolutionary compensation of uORFs in Drosophila Jian Lu, Chenlu Liu, Wanting Jin, Yuanqiang Sun, Yuange Duan, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4896906/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 Upstream open reading frames (uORFs) play crucial roles in regulating mRNA translation; however, the evolutionary dynamics of uORFs and the potential coevolution of uORF gain and loss remain largely unexplored. Here, we comprehensively analyze the evolutionary landscape of uORFs across Drosophila species, revealing a complex pattern of birth and death events throughout the phylogeny, with a net gain trend dominating the overall pattern. We identified instances of evolutionary compensation in which the loss of a uORF was accompanied by the gain of another within the same gene, or vice versa. Surprisingly, a significant proportion of fixed uORF loss events in D. melanogaster were driven by positive selection. Dual-luciferase assays revealed that approximately 75% of the tested uORF gains or losses altered translational regulation, with some exhibiting condition-specific effects. Surveying uORF-related variants across 1,356 D. melanogaster genomes revealed strong purifying selection, with some variants potentially contributing to local adaptation. Our findings highlight the crucial role of natural selection in shaping the evolutionary landscape of uORFs in Drosophila and provide novel insights into the complex interplay between the evolution of coding and regulatory elements. Biological sciences/Evolution/Molecular evolution Biological sciences/Evolution/Evolutionary genetics Upstream open reading frames (uORFs) birth and death translational regulation natural selection compensatory evolution Drosophila Full Text Additional Declarations There is NO Competing Interest. Supplementary Files uORFbirthdeathSOM20240811NEE.pdf tablesuORF0809NEE.xlsx 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. 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-4896906","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":346343864,"identity":"79c58d31-19cf-4633-8653-0826dbcb65aa","order_by":0,"name":"Jian 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