Architecture and self-assembly of the SARS-CoV-2 nucleocapsid protein

BioRxiv : the Preprint Server for Biology
Q. YeKevin D Corbett

Abstract

The COVID-2019 pandemic is the most severe acute public health threat of the twenty-first century. To properly address this crisis with both robust testing and novel treatments, we require a deep understanding of the life cycle of the causative agent, the SARS-CoV-2 coronavirus. Here, we examine the architecture and self-assembly properties of the SARS-CoV-2 nucleocapsid (N) protein, which binds viral RNA and assembles into a filament that is packaged into new virions. We determined a 1.4 angstrom resolution crystal structure of this protein's N2b domain, revealing a compact, intertwined dimer very similar to that of related coronaviruses SARS-CoV and MERS-CoV. Using size exclusion chromatography and multi-angle light scattering, we find that this domain forms a dimer in solution, and that addition of the C-terminal spacer B/N3 domain mediates tetramer formation. Using hydrogen-deuterium exchange mass spectrometry, we find evidence that at least part of this putatively disordered domain is structured, potentially forming an -helix that either self-associates or docks against the N2b domain to mediate tetramer formation. Finally, we map the locations of over 4,400 individual amino acid substitutions in the N protein from ~17,000...Continue Reading

Methods Mentioned

BETA
two-hybrid
NMR
gel filtration
Size exclusion chromatography
Size
light scattering
PCR
size-exclusion
light

Software Mentioned

PRED
PHASER
phenix
Prism
DECA
CCP4
COOT
Jalview
XDS
TRUNCATE

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