Structures of Natural and Strategic Mutants reveal a Core Network crucial for Integrity of the SARS-CoV-2 Nucleocapsid NTD

Structures of Natural and Strategic Mutants reveal a Core Network crucial for Integrity of the SARS-CoV-2 Nucleocapsid NTD | 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 Structures of Natural and Strategic Mutants reveal a Core Network crucial for Integrity of the SARS-CoV-2 Nucleocapsid NTD Andreas Schlundt, Karthikeyan Dhamotharan, Sophie Korn, Anna Wacker, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4595915/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Dec, 2024 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract The SARS-CoV-2 nucleocapsid protein is indispensable for processing of the viral RNA genome. The N-terminal domain (NTD) was suggested to mediate specific RNA-interactions, but the lack of high-resolution structures with viral RNA prohibits insights into the precise mechanism of complex formation. Similarly, mutations in the nucleocapsid NTD since 2019 have not been addressed molecularly. Systematically combining crystallography and solution NMR, we here investigate the influence of six naturally occurring plus strategic NTD mutations on structural integrity and RNA-binding. We find that both features rely on a core network of residues conserved in Betacoronaviruses. This core network proves important for protein stability and communication between flexible loop-regions, key for RNA-recognition. Our comprehensive structural analysis demonstrates that crucial contacts within the expanded network steer selective interaction with target RNAs. We propose the core network renders the NTD evolutionarily robust in stability and plasticity for its versatile RNA genome-processing functions. Biological sciences/Structural biology/X-ray crystallography Biological sciences/Biochemistry/Structural biology/NMR spectroscopy/Solution-state NMR Biological sciences/Biochemistry/Proteins/RNA-binding proteins SARS-CoV-2 Nucleocapsid RNA-binding protein Variants of concern (VOC) X-ray structures Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformationDhamotharanetal.pdf Cite Share Download PDF Status: Published Journal Publication published 09 Dec, 2024 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. 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