tetano
Editor, Senior Moderator
Biochem Biophys Res Commun
. 2020 Oct 3;S0006-291X(20)31884-2.
doi: 10.1016/j.bbrc.2020.09.131. Online ahead of print.
High-resolution structure and biophysical characterization of the nucleocapsid phosphoprotein dimerization domain from the Covid-19 severe acute respiratory syndrome coronavirus 2
Luca Zinzula[SUP] 1 [/SUP], Jerome Basquin[SUP] 2 [/SUP], Stefan Bohn[SUP] 3 [/SUP], Florian Beck[SUP] 4 [/SUP], Sven Klumpe[SUP] 4 [/SUP], G?nter Pfeifer[SUP] 4 [/SUP], Istv?n Nagy[SUP] 4 [/SUP], Andreas Bracher[SUP] 5 [/SUP], F Ulrich Hartl[SUP] 5 [/SUP], Wolfgang Baumeister[SUP] 6 [/SUP]
Affiliations
Abstract
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 ? and 1.36 ? 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.
Keywords: Covid-19; Nucleocapsid; Oligomerization; RNA binding; SARS coronavirus.
. 2020 Oct 3;S0006-291X(20)31884-2.
doi: 10.1016/j.bbrc.2020.09.131. Online ahead of print.
High-resolution structure and biophysical characterization of the nucleocapsid phosphoprotein dimerization domain from the Covid-19 severe acute respiratory syndrome coronavirus 2
Luca Zinzula[SUP] 1 [/SUP], Jerome Basquin[SUP] 2 [/SUP], Stefan Bohn[SUP] 3 [/SUP], Florian Beck[SUP] 4 [/SUP], Sven Klumpe[SUP] 4 [/SUP], G?nter Pfeifer[SUP] 4 [/SUP], Istv?n Nagy[SUP] 4 [/SUP], Andreas Bracher[SUP] 5 [/SUP], F Ulrich Hartl[SUP] 5 [/SUP], Wolfgang Baumeister[SUP] 6 [/SUP]
Affiliations
- PMID: 33039147
- PMCID: PMC7532810
- DOI: 10.1016/j.bbrc.2020.09.131
Abstract
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 ? and 1.36 ? 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.
Keywords: Covid-19; Nucleocapsid; Oligomerization; RNA binding; SARS coronavirus.