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Structural basis of RNA recognition by the SARS-CoV-2 nucleocapsid phosphoprotein
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SYSNO ASEP 0536797 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Structural basis of RNA recognition by the SARS-CoV-2 nucleocapsid phosphoprotein Author(s) Dinesh, Dhurvas Chandrasekaran (UOCHB-X) ORCID
Chalupská, Dominika (UOCHB-X) ORCID
Šilhán, Jan (UOCHB-X) ORCID
Koutná, Eliška (UOCHB-X) ORCID
Nencka, Radim (UOCHB-X) RID, ORCID
Veverka, Václav (UOCHB-X) RID, ORCID
Bouřa, Evžen (UOCHB-X) ORCIDArticle number e1009100 Source Title PLoS Pathogens. - : Public Library of Science - ISSN 1553-7366
Roč. 16, č. 12 (2020)Number of pages 16 s. Language eng - English Country US - United States Keywords N-terminal domain ; binding domain ; crystal structure Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology R&D Projects EF16_019/0000729 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000596505200002 EID SCOPUS 85097516875 DOI 10.1371/journal.ppat.1009100 Annotation Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease 2019 (COVID-19). SARS-CoV-2 is a single-stranded positivesense RNA virus. Like other coronaviruses, SARS-CoV-2 has an unusually large genome that encodes four structural proteins and sixteen nonstructural proteins. The structural nucleocapsid phosphoprotein N is essential for linking the viral genome to the viral membrane. Both N-terminal RNA binding (N-NTD) and C-terminal dimerization domains are involved in capturing the RNA genome and, the intrinsically disordered region between these domains anchors the ribonucleoprotein complex to the viral membrane. Here, we characterized the structure of the N-NTD and its interaction with RNA using NMR spectroscopy. We observed a positively charged canyon on the surface of the N-NTD that might serve as a putative RNA binding site similarly to other coronaviruses. The subsequent NMR titrations using single-stranded and double-stranded RNA revealed a much more extensive U-shaped RNA-binding cleft lined with regularly distributed arginines and lysines. The NMR data supported by mutational analysis allowed us to construct hybrid atomic models of the N-NTD/RNA complex that provided detailed insight into RNA recognition. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2021 Electronic address https://doi.org/10.1371/journal.ppat.1009100
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