High-resolution structure and biophysical characterization of the nucleocapsid phosphoprotein dimerization domain from the Covid-19 severe acute respiratory syndrome coronavirus 2
| Autor: | F. Ulrich Hartl, Luca Zinzula, Andreas Bracher, István Nagy, Stefan Bohn, Florian Beck, Sven Klumpe, Günter Pfeifer, Wolfgang Baumeister, Jérôme Basquin |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Conformational change SARS coronavirus viruses Protein domain Biophysics Virulence RNA-binding protein Plasma protein binding Genome Viral Biochemistry Article 03 medical and health sciences 0302 clinical medicine Protein Domains Oligomerization Coronavirus Nucleocapsid Proteins Humans Nucleocapsid Molecular Biology Microscale thermophoresis Chemistry SARS-CoV-2 Cryoelectron Microscopy RNA COVID-19 RNA-Binding Proteins Cell Biology RNA binding Phosphoproteins 030104 developmental biology 030220 oncology & carcinogenesis Phosphoprotein Protein Multimerization Protein Binding |
| Zdroj: | Biochemical and Biophysical Research Communications |
| ISSN: | 1090-2104 0006-291X |
| Popis: | Unprecedented by number of casualties and socio-economic burden occurring worldwide, the coronavirus disease 2019 (Covid-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the worst health crisis of this century. In order to develop adequate countermeasures against Covid-19, identification and structural characterization of suitable antiviral targets within the SARS-CoV-2 protein repertoire is urgently needed. The nucleocapsid phosphoprotein (N) is a multifunctional and highly immunogenic determinant of virulence and pathogenicity, whose main functions consist in oligomerizing and packaging the single-stranded RNA (ssRNA) viral genome. Here we report the structural and biophysical characterization of the SARS-CoV-2 N C-terminal domain (CTD), on which both N homo-oligomerization and ssRNA binding depend. Crystal structures solved at 1.44 Å and 1.36 Å resolution describe a rhombus-shape N CTD dimer, which stably exists in solution as validated by size-exclusion chromatography coupled to multi-angle light scattering and analytical ultracentrifugation. Differential scanning fluorimetry revealed moderate thermal stability and a tendency towards conformational change. Microscale thermophoresis demonstrated binding to a 7-bp SARS-CoV-2 genomic ssRNA fragment at micromolar affinity. Furthermore, a low-resolution preliminary model of the full-length SARS-CoV N in complex with ssRNA, obtained by cryo-electron microscopy, provides an initial understanding of self-associating and RNA binding functions exerted by the SARS-CoV-2 N. Highlights • SARS-CoV-2 nucleocapsid phosphoprotein (N) CTD forms a stable dimer in solution. • SARS-CoV-2 N CTD dimer is a rhombus-shaped tile similar to SARS-CoV and MERS-CoV. • SARS-CoV-2 N CTD binds to a 7bp viral genomic ssRNA with micromolar affinity. • SARS-CoV-2 N CTD structure provides a framework for drug design against Covid-19. |
| Databáze: | OpenAIRE |
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