+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Coronavirus particle assembly: primary structure requirements of the membrane protein

Coronavirus particle assembly: primary structure requirements of the membrane protein

Journal of Virology 72(8): 6838-6850

Coronavirus-like particles morphologically similar to normal virions are assembled when genes encoding the viral membrane proteins M and E are coexpressed in eukaryotic cells. Using this envelope assembly assay, we have studied the primary sequence requirements for particle formation of the mouse hepatitis virus (MHV) M protein, the major protein of the coronavirion membrane. Our results show that each of the different domains of the protein is important. Mutations (deletions, insertions, point mutations) in the luminal domain, the transmembrane domains, the amphiphilic domain, or the carboxy-terminal domain had effects on the assembly of M into enveloped particles. Strikingly, the extreme carboxy-terminal residue is crucial. Deletion of this single residue abolished particle assembly almost completely; most substitutions were strongly inhibitory. Site-directed mutations in the carboxy terminus of M were also incorporated into the MHV genome by targeted recombination. The results supported a critical role for this domain of M in viral assembly, although the M carboxy terminus was more tolerant of alteration in the complete virion than in virus-like particles, likely because of the stabilization of virions by additional intermolecular interactions. Interestingly, glycosylation of M appeared not essential for assembly. Mutations in the luminal domain that abolished the normal O glycosylation of the protein or created an N-glycosylated form had no effect. Mutant M proteins unable to form virus-like particles were found to inhibit the budding of assembly-competent M in a concentration-dependent manner. However, assembly-competent M was able to rescue assembly-incompetent M when the latter was present in low amounts. These observations support the existence of interactions between M molecules that are thought to be the driving force in coronavirus envelope assembly.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 008391186

Download citation: RISBibTeXText

PMID: 9658133

Related references

Hepatitis B virus nucleocapsid assembly: primary structure requirements in the core protein. Journal of Virology 64(7): 3319-3330, 1990

The missing link in coronavirus assembly. Retention of the avian coronavirus infectious bronchitis virus envelope protein in the pre-Golgi compartments and physical interaction between the envelope and membrane proteins. Journal of Biological Chemistry 276(20): 17515-17523, 2001

Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome. Proceedings of the National Academy of Sciences of the United States of America 108(7): 2759-2764, 2011

In vitro assembly of the murine coronavirus membrane protein E1. Advances in Experimental Medicine and Biology 173: 53-64, 1984

Evolved variants of the membrane protein can partially replace the envelope protein in murine coronavirus assembly. Journal of Virology 84(24): 12872-12885, 2010

Self-assembly of severe acute respiratory syndrome coronavirus membrane protein. Journal of Biological Chemistry 285(17): 12862-12872, 2010

A conserved domain in the coronavirus membrane protein tail is important for virus assembly. Journal of Virology 84(21): 11418-11428, 2010

Analyses of Coronavirus Assembly Interactions with Interspecies Membrane and Nucleocapsid Protein Chimeras. Journal of Virology 90(9): 4357-4368, 2016

A minimized M13 coat protein defines the requirements for assembly into the bacteriophage particle. Journal of Molecular Biology 322(2): 7-67, 2002

Mutation in murine coronavirus replication protein nsp4 alters assembly of double membrane vesicles. Virology 375(1): 118-129, 2008

Assembly in vitro of a spanning membrane protein of the endoplasmic reticulum: the E1 glycoprotein of coronavirus mouse hepatitis virus A59. Proceedings of the National Academy of Sciences of the United States of America 81(5): 1421-1425, 1984

Membrane assembly of the triple-spanning coronavirus, protein: Individual transmembrane domains show preferred orientation. Journal of Biological Chemistry 267(30): 21911-21918, 1992

Interaction of the coronavirus infectious bronchitis virus membrane protein with beta-actin and its implication in virion assembly and budding. Plos one 4(3): E4908, 2009