Coronavirus nucleocapsid proteins assemble constitutively in high molecular oligomers

Scientific Reports
Yingying CongFulvio Reggiori

Abstract

Coronaviruses (CoV) are enveloped viruses and rely on their nucleocapsid N protein to incorporate the positive-stranded genomic RNA into the virions. CoV N proteins form oligomers but the mechanism and relevance underlying their multimerization remain to be fully understood. Using in vitro pull-down experiments and density glycerol gradients, we found that at least 3 regions distributed over its entire length mediate the self-interaction of mouse hepatitis virus (MHV) and severe acute respiratory syndrome coronavirus (SARS-CoV) N protein. The fact that these regions can bind reciprocally between themselves provides a possible molecular basis for N protein oligomerization. Interestingly, cytoplasmic N molecules of MHV-infected cells constitutively assemble into oligomers through a process that does not require binding to genomic RNA. Based on our data, we propose a model where constitutive N protein oligomerization allows the optimal loading of the genomic viral RNA into a ribonucleoprotein complex via the presentation of multiple viral RNA binding motifs.

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Citations

Apr 17, 2020·Journal of Biomolecular Structure & Dynamics·Maryam EnayatkhaniKhadijeh Ahmadi
Jun 20, 2020·Journal of Medical Virology·Mahdi Muhammad Moosa, Priya R Banerjee
Jul 13, 2020·Protein Science : a Publication of the Protein Society·Qiaozhen YeKevin D Corbett
Oct 10, 2020·Cell and Tissue Banking·André Oliveira PaggiaroRolf Gemperli
Nov 22, 2020·Analytical and Bioanalytical Chemistry·Mercy R BenzigarGuozhen Liu
Nov 18, 2020·The EMBO Journal·Theodora Myrto PerdikariNicolas L Fawzi
Oct 13, 2019·Seminars in Immunology·Walter M KimRichard S Blumberg
Aug 25, 2020·Heliyon·I Made ArtikaAgeng Wiyatno
Dec 9, 2020·Molecular Cell·Christiane IsermanAmy S Gladfelter
Feb 19, 2021·Biology of the Cell·Lucio Ayres CaldasWanderley de Souza
Mar 18, 2021·Nature Reviews Physics·Robijn F BruinsmaWouter H Roos
Mar 31, 2021·Nature Communications·Jasmine CubukAlex S Holehouse

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Methods Mentioned

BETA
two-hybrid
co-immunoprecipitation
size
surface plasmon resonance
pull-down
PCR
pull-downs

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