Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures
Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures
Summary Betacoronavirus SARS-CoV-2 infections caused the global Covid-19 pandemic. The nucleocapsid protein (N-protein) is required for multiple steps in the betacoronavirus replication cycle. SARS-CoV-2-N-protein is known to undergo liquid-liquid phase separation (LLPS) with specific RNAs at particular temperatures to form condensates. We show that N-protein recognizes at least two separate and distinct RNA motifs, both of which require double-stranded RNA (dsRNA) for LLPS. These motifs are separately recognized by N-protein’s two RNA binding domains (RBDs). Addition of dsRNA accelerates and modifies N-protein LLPS in vitro and in cells and controls the temperature condensates form. The abundance of dsRNA tunes N-protein-mediated translational repression and may confer a switch from translation to genome packaging. Thus, N-protein’s two RBDs interact with separate dsRNA motifs, and these interactions impart distinct droplet properties that can support multiple viral functions. These experiments demonstrate a paradigm of how RNA structure can control the properties of biomolecular condensates.
Ekena Joanne L.、Chilkoti Ashutosh、Lee Myungwoon、McLaughlin Grace A.、Weeks Kevin M.、Roden Christine A.、Sealfon Rachel、Seim Ian、Wey Samuel A.、Troyanskaya Olga G.、Gladfelter Amy S.、Iserman Christiane、Boerneke Mark A.、You Lingchong、Dai Yifan
Department of Biology, University of North Carolina at Chapel HillDepartment of Biomedical Engineering, Duke UniversityLaboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthDepartment of Biology, University of North Carolina at Chapel HillDepartment of Chemistry, University of North Carolina at Chapel HillDepartment of Biology, University of North Carolina at Chapel Hill||Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel HillFlatiron Institute, Simons FoundationDepartment of Biology, University of North Carolina at Chapel Hill||Curriculum in Bioinformatics and Computational Biology, University of North Carolina at Chapel Hill||Department of Applied Physical Sciences, University of North Carolina at Chapel HillDepartment of Chemistry, University of North Carolina at Chapel HillFlatiron Institute, Simons Foundation||Department of Computer Science, Princeton University||Lewis-Sigler Institute for Integrative Genomics, Princeton UniversityDepartment of Biology, University of North Carolina at Chapel Hill||Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel HillDepartment of Biology, University of North Carolina at Chapel HillDepartment of Chemistry, University of North Carolina at Chapel HillDepartment of Biomedical Engineering, Duke University||Center for Genomic and Computational Biology, Duke University||Department of Molecular Genetics and Microbiology, Duke University School of MedicineDepartment of Biomedical Engineering, Duke University
分子生物学生物化学基础医学
SARS-CoV-2nucleocapsidliquid-liquid phase separation (LLPS)double-stranded RNA (dsRNA)RNA stickers and spacerslower critical solution temperature (LCST)translational repressionviral genome packagingRNP formation.
Ekena Joanne L.,Chilkoti Ashutosh,Lee Myungwoon,McLaughlin Grace A.,Weeks Kevin M.,Roden Christine A.,Sealfon Rachel,Seim Ian,Wey Samuel A.,Troyanskaya Olga G.,Gladfelter Amy S.,Iserman Christiane,Boerneke Mark A.,You Lingchong,Dai Yifan.Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures[EB/OL].(2025-03-28)[2025-08-02].https://www.biorxiv.org/content/10.1101/2021.06.14.448452.点此复制
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