Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: implications for assembly and vaccine production.
Identifieur interne : 002A77 ( PubMed/Curation ); précédent : 002A76; suivant : 002A78Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: implications for assembly and vaccine production.
Auteurs : Yue Huang [États-Unis] ; Zhi-Yong Yang ; Wing-Pui Kong ; Gary J. NabelSource :
- Journal of virology [ 0022-538X ] ; 2004.
Descripteurs français
- KwdFr :
- Assemblage viral, Données de séquences moléculaires, Gènes viraux (physiologie), Humains, Nucléocapside (physiologie), Protéines de la matrice virale (génétique), Protéines nucléocapside (génétique), Séquence nucléotidique, Vaccins antiviraux (immunologie), Virion (physiologie), Virus du SRAS (génétique), Virus du SRAS (immunologie), Virus du SRAS (physiologie).
- MESH :
- génétique : Protéines de la matrice virale, Protéines nucléocapside, Virus du SRAS.
- immunologie : Vaccins antiviraux, Virus du SRAS.
- physiologie : Gènes viraux, Nucléocapside, Virion, Virus du SRAS.
- Assemblage viral, Données de séquences moléculaires, Humains, Séquence nucléotidique.
English descriptors
- KwdEn :
- Base Sequence, Genes, Viral (physiology), Humans, Molecular Sequence Data, Nucleocapsid (physiology), Nucleocapsid Proteins (genetics), SARS Virus (genetics), SARS Virus (immunology), SARS Virus (physiology), Viral Matrix Proteins (genetics), Viral Vaccines (immunology), Virion (physiology), Virus Assembly.
- MESH :
- chemical , genetics : Nucleocapsid Proteins, Viral Matrix Proteins.
- genetics : SARS Virus.
- immunology : SARS Virus, Viral Vaccines.
- physiology : Genes, Viral, Nucleocapsid, SARS Virus, Virion.
- Base Sequence, Humans, Molecular Sequence Data, Virus Assembly.
Abstract
The recently emerged severe acute respiratory syndrome coronavirus (SARS-CoV) contains four structural genes, two replicase-transcriptase open reading frames, and more than five potential genes of unknown function. Despite this relative simplicity, the molecular regulation of SARS-CoV replication and assembly is not understood. Here, we report that two viral genes, encoding the SARS-CoV membrane (M) and nucleocapsid (N) proteins, are necessary and sufficient for formation of virus-like particles. Expression vectors encoding these two proteins were synthesized by using preferred human codons. When M and N expression plasmids were cotransfected into human 293 renal epithelial cells, pseudoparticles formed readily. The addition of a third gene, encoding the spike (S) glycoprotein, facilitated budding of particles that contained a corona-like halo resembling SARS-CoV when examined by transmission electron microscopy, with a buoyant density characteristic of coronaviruses. Specific biochemical interactions of these proteins were also shown in vitro. The S, M, and N proteins of the SARS-CoV are, therefore, necessary and sufficient for pseudovirus assembly. These findings advance the understanding of the morphogenesis of SARS-CoV and enable the generation of safe, conformational mimetics of the SARS virus that may facilitate the development of vaccines and antiviral drugs.
DOI: 10.1128/JVI.78.22.12557-12565.2004
PubMed: 15507643
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<front><div type="abstract" xml:lang="en">The recently emerged severe acute respiratory syndrome coronavirus (SARS-CoV) contains four structural genes, two replicase-transcriptase open reading frames, and more than five potential genes of unknown function. Despite this relative simplicity, the molecular regulation of SARS-CoV replication and assembly is not understood. Here, we report that two viral genes, encoding the SARS-CoV membrane (M) and nucleocapsid (N) proteins, are necessary and sufficient for formation of virus-like particles. Expression vectors encoding these two proteins were synthesized by using preferred human codons. When M and N expression plasmids were cotransfected into human 293 renal epithelial cells, pseudoparticles formed readily. The addition of a third gene, encoding the spike (S) glycoprotein, facilitated budding of particles that contained a corona-like halo resembling SARS-CoV when examined by transmission electron microscopy, with a buoyant density characteristic of coronaviruses. Specific biochemical interactions of these proteins were also shown in vitro. The S, M, and N proteins of the SARS-CoV are, therefore, necessary and sufficient for pseudovirus assembly. These findings advance the understanding of the morphogenesis of SARS-CoV and enable the generation of safe, conformational mimetics of the SARS virus that may facilitate the development of vaccines and antiviral drugs.</div>
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