The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.
Identifieur interne : 003064 ( Main/Merge ); précédent : 003063; suivant : 003065The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.
Auteurs : Y L Siu [République populaire de Chine] ; K T Teoh ; J. Lo ; C M Chan ; F. Kien ; N. Escriou ; S W Tsao ; J M Nicholls ; R. Altmeyer ; J S M. Peiris ; R. Bruzzone ; B. NalSource :
- Journal of virology [ 1098-5514 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Assemblage viral, Cellules Vero, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (métabolisme), Humains, Microscopie électronique à transmission, Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (métabolisme), Protéines de la matrice virale (génétique), Protéines de la matrice virale (métabolisme), Protéines nucléocapside (génétique), Protéines nucléocapside (métabolisme), Virosomes (métabolisme), Virosomes (ultrastructure), Virus du SRAS (physiologie).
- MESH :
- génétique : Protéines de l'enveloppe virale, Protéines de la matrice virale, Protéines nucléocapside.
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines de la matrice virale, Protéines nucléocapside, Virosomes.
- physiologie : Virus du SRAS.
- ultrastructure : Animaux, Assemblage viral, Cellules Vero, Glycoprotéine de spicule des coronavirus, Humains, Microscopie électronique à transmission, Virosomes.
English descriptors
- KwdEn :
- Animals, Chlorocebus aethiops, Humans, Membrane Glycoproteins (metabolism), Microscopy, Electron, Transmission, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (metabolism), SARS Virus (physiology), Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins (genetics), Viral Envelope Proteins (metabolism), Viral Matrix Proteins (genetics), Viral Matrix Proteins (metabolism), Virosomes (metabolism), Virosomes (ultrastructure), Virus Assembly.
- MESH :
- chemical , genetics : Nucleocapsid Proteins, Viral Envelope Proteins, Viral Matrix Proteins.
- chemical , metabolism : Membrane Glycoproteins, Nucleocapsid Proteins, Viral Envelope Proteins, Viral Matrix Proteins, Virosomes.
- physiology : SARS Virus.
- chemical , ultrastructure : Virosomes.
- Animals, Chlorocebus aethiops, Humans, Microscopy, Electron, Transmission, Spike Glycoprotein, Coronavirus, Vero Cells, Virus Assembly.
Abstract
The production of virus-like particles (VLPs) constitutes a relevant and safe model to study molecular determinants of virion egress. The minimal requirement for the assembly of VLPs for the coronavirus responsible for severe acute respiratory syndrome in humans (SARS-CoV) is still controversial. Recent studies have shown that SARS-CoV VLP formation depends on either M and E proteins or M and N proteins. Here we show that both E and N proteins must be coexpressed with M protein for the efficient production and release of VLPs by transfected Vero E6 cells. This suggests that the mechanism of SARS-CoV assembly differs from that of other studied coronaviruses, which only require M and E proteins for VLP formation. When coexpressed, the native envelope trimeric S glycoprotein is incorporated onto VLPs. Interestingly, when a fluorescent protein tag is added to the C-terminal end of N or S protein, but not M protein, the chimeric viral proteins can be assembled within VLPs and allow visualization of VLP production and trafficking in living cells by state-of-the-art imaging technologies. Fluorescent VLPs will be used further to investigate the role of cellular machineries during SARS-CoV egress.
Url:
DOI: 10.1128/JVI.01052-08
PubMed: 18753196
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Links to Exploration step
pubmed:18753196Le document en format XML
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The minimal requirement for the assembly of VLPs for the coronavirus responsible for severe acute respiratory syndrome in humans (SARS-CoV) is still controversial. Recent studies have shown that SARS-CoV VLP formation depends on either M and E proteins or M and N proteins. Here we show that both E and N proteins must be coexpressed with M protein for the efficient production and release of VLPs by transfected Vero E6 cells. This suggests that the mechanism of SARS-CoV assembly differs from that of other studied coronaviruses, which only require M and E proteins for VLP formation. When coexpressed, the native envelope trimeric S glycoprotein is incorporated onto VLPs. Interestingly, when a fluorescent protein tag is added to the C-terminal end of N or S protein, but not M protein, the chimeric viral proteins can be assembled within VLPs and allow visualization of VLP production and trafficking in living cells by state-of-the-art imaging technologies. Fluorescent VLPs will be used further to investigate the role of cellular machineries during SARS-CoV egress.</div></front></TEI><double doi="10.1128/JVI.01052-08"><HAL><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.</title><author><name sortKey="Siu, Y L" sort="Siu, Y L" uniqKey="Siu Y" first="Y. L." last="Siu">Y. L. 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M." last="Chan">C. M. Chan</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-136023" status="VALID"> <orgName>Department of Microbiology [HKU]</orgName> <desc> <address> <addrLine>Hong Kong Special Administrative Region, China</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Kien, F" sort="Kien, F" uniqKey="Kien F" first="F." last="Kien">F. 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W." last="Tsao">S. W. Tsao</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-135181" status="VALID"> <orgName>Department of Anatomy [HKU]</orgName> <desc> <address> <addrLine>Hong Kong</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Nicholls, J M" sort="Nicholls, J M" uniqKey="Nicholls J" first="J. M." last="Nicholls">J. M. Nicholls</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-157371" status="VALID"> <orgName>Department of pathology [HKU]</orgName> <desc> <address> <addrLine>21 Sassoon Road, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong Special Administrative Region</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Altmeyer, R" sort="Altmeyer, R" uniqKey="Altmeyer R" first="R." last="Altmeyer">R. Altmeyer</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-135182" status="INCOMING"> <orgName>CombinatoRx</orgName> <desc> <address> <addrLine>Singapour</addrLine> <country key="SG"></country> </address> </desc> <listRelation> <relation active="#struct-332787" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-332787" type="direct"><org type="institution" xml:id="struct-332787" status="INCOMING"> <orgName>CombinatoRx</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle></tutelles></hal:affiliation><country>Singapour</country></affiliation></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J. S. M." last="Peiris">J. S. M. Peiris</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Bruzzone, R" sort="Bruzzone, R" uniqKey="Bruzzone R" first="R." last="Bruzzone">R. Bruzzone</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Nal, B" sort="Nal, B" uniqKey="Nal B" first="B." last="Nal">B. Nal</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">HAL</idno><idno type="RBID">Hal:pasteur-00543224</idno><idno type="halId">pasteur-00543224</idno><idno type="halUri">https://hal-riip.archives-ouvertes.fr/pasteur-00543224</idno><idno type="url">https://hal-riip.archives-ouvertes.fr/pasteur-00543224</idno><idno type="doi">10.1128/JVI.01052-08</idno><date when="2008-11">2008-11</date><idno type="wicri:Area/Hal/Corpus">000159</idno><idno type="wicri:Area/Hal/Curation">000159</idno><idno type="wicri:Area/Hal/Checkpoint">000151</idno><idno type="wicri:explorRef" wicri:stream="Hal" wicri:step="Checkpoint">000151</idno><idno type="wicri:doubleKey">0022-538X:2008:Siu Y:the:m:e</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.</title><author><name sortKey="Siu, Y L" sort="Siu, Y L" uniqKey="Siu Y" first="Y. L." last="Siu">Y. L. Siu</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Teoh, K T" sort="Teoh, K T" uniqKey="Teoh K" first="K. T." last="Teoh">K. T. Teoh</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Lo, J" sort="Lo, J" uniqKey="Lo J" first="J." last="Lo">J. Lo</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-157371" status="VALID"> <orgName>Department of pathology [HKU]</orgName> <desc> <address> <addrLine>21 Sassoon Road, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong Special Administrative Region</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Chan, C M" sort="Chan, C M" uniqKey="Chan C" first="C. M." last="Chan">C. M. Chan</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-136023" status="VALID"> <orgName>Department of Microbiology [HKU]</orgName> <desc> <address> <addrLine>Hong Kong Special Administrative Region, China</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Kien, F" sort="Kien, F" uniqKey="Kien F" first="F." last="Kien">F. Kien</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Escriou, N" sort="Escriou, N" uniqKey="Escriou N" first="N." last="Escriou">N. Escriou</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-31889" status="OLD"> <orgName>Génétique Moléculaire des Virus Respiratoires</orgName> <desc> <address> <addrLine>25-28 rue du Docteur Roux, F-75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> </desc> <listRelation> <relation active="#struct-300027" type="direct"></relation> <relation active="#struct-300301" type="direct"></relation> <relation name="URA1966" active="#struct-441569" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-300027" type="direct"><org type="institution" xml:id="struct-300027" status="VALID"> <idno type="IdRef">027936643</idno> <idno type="ISNI">0000 0001 2353 6535</idno> <orgName>Institut Pasteur [Paris]</orgName> <date type="start">1887-06-04</date> <desc> <address> <addrLine>25-28, rue du docteur Roux, 75724 Paris cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://www.pasteur.fr</ref> </desc> </org></tutelle><tutelle active="#struct-300301" type="direct"><org type="institution" xml:id="struct-300301" status="OLD"> <idno type="ISNI">0000000121514068</idno> <idno type="IdRef">027542084</idno> <orgName>Université Paris Diderot - Paris 7</orgName> <orgName type="acronym">UPD7</orgName> <date type="end">2019-12-31</date> <desc> <address> <addrLine>5 rue Thomas-Mann - 75205 Paris cedex 13</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.univ-paris-diderot.fr</ref> </desc> </org></tutelle><tutelle name="URA1966" active="#struct-441569" type="direct"><org type="institution" xml:id="struct-441569" status="VALID"> <idno type="IdRef">02636817X</idno> <idno type="ISNI">0000000122597504</idno> <orgName>Centre National de la Recherche Scientifique</orgName> <orgName type="acronym">CNRS</orgName> <date type="start">1939-10-19</date> <desc> <address> <country key="FR"></country> </address> <ref type="url">http://www.cnrs.fr/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Tsao, S W" sort="Tsao, S W" uniqKey="Tsao S" first="S. W." last="Tsao">S. W. Tsao</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-135181" status="VALID"> <orgName>Department of Anatomy [HKU]</orgName> <desc> <address> <addrLine>Hong Kong</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Nicholls, J M" sort="Nicholls, J M" uniqKey="Nicholls J" first="J. M." last="Nicholls">J. M. Nicholls</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-157371" status="VALID"> <orgName>Department of pathology [HKU]</orgName> <desc> <address> <addrLine>21 Sassoon Road, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong Special Administrative Region</addrLine> <country key="CN"></country> </address> </desc> <listRelation> <relation active="#struct-93487" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-93487" type="direct"><org type="institution" xml:id="struct-93487" status="VALID"> <orgName>The University of Hong Kong</orgName> <orgName type="acronym">HKU</orgName> <desc> <address> <addrLine>Pokfulam, Hong Kong</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hku.hk/</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Altmeyer, R" sort="Altmeyer, R" uniqKey="Altmeyer R" first="R." last="Altmeyer">R. Altmeyer</name><affiliation wicri:level="1"><hal:affiliation type="laboratory" xml:id="struct-135182" status="INCOMING"> <orgName>CombinatoRx</orgName> <desc> <address> <addrLine>Singapour</addrLine> <country key="SG"></country> </address> </desc> <listRelation> <relation active="#struct-332787" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-332787" type="direct"><org type="institution" xml:id="struct-332787" status="INCOMING"> <orgName>CombinatoRx</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle></tutelles></hal:affiliation><country>Singapour</country></affiliation></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J. S. M." last="Peiris">J. S. M. Peiris</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Bruzzone, R" sort="Bruzzone, R" uniqKey="Bruzzone R" first="R." last="Bruzzone">R. Bruzzone</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author><author><name sortKey="Nal, B" sort="Nal, B" uniqKey="Nal B" first="B." last="Nal">B. Nal</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-55925" status="VALID"> <orgName>Centre de recherche Université de Hong-Kong-Pasteur</orgName> <desc> <address> <addrLine>Hong Kong University Pasteur Research Centre - 1/F Dexter HC Man Building 8, Sassoon Road Pokfulam</addrLine> <country key="HK"></country> </address> <ref type="url">http://www.hkupasteur.hku.hk/</ref> </desc> <listRelation> <relation active="#struct-301247" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-301247" type="direct"><org type="regroupinstitution" xml:id="struct-301247" status="VALID"> <orgName>Réseau International des Instituts Pasteur</orgName> <orgName type="acronym">RIIP</orgName> <desc> <address> <addrLine>25, rue du Dr Roux 75724 Paris Cedex 15</addrLine> <country key="FR"></country> </address> <ref type="url">https://research.pasteur.fr/ip-network</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>Hong Kong</country></affiliation></author></analytic><idno type="DOI">10.1128/JVI.01052-08</idno><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><imprint><date type="datePub">2008-11</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"> <p>The production of virus-like particles (VLPs) constitutes a relevant and safe model to study molecular determinants of virion egress. The minimal requirement for the assembly of VLPs for the coronavirus responsible for severe acute respiratory syndrome in humans (SARS-CoV) is still controversial. Recent studies have shown that SARS-CoV VLP formation depends on either M and E proteins or M and N proteins. Here we show that both E and N proteins must be coexpressed with M protein for the efficient production and release of VLPs by transfected Vero E6 cells. This suggests that the mechanism of SARS-CoV assembly differs from that of other studied coronaviruses, which only require M and E proteins for VLP formation. When coexpressed, the native envelope trimeric S glycoprotein is incorporated onto VLPs. Interestingly, when a fluorescent protein tag is added to the C-terminal end of N or S protein, but not M protein, the chimeric viral proteins can be assembled within VLPs and allow visualization of VLP production and trafficking in living cells by state-of-the-art imaging technologies. Fluorescent VLPs will be used further to investigate the role of cellular machineries during SARS-CoV egress.</p> </div></front></TEI></HAL><PubMed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.</title><author><name sortKey="Siu, Y L" sort="Siu, Y L" uniqKey="Siu Y" first="Y L" last="Siu">Y L Siu</name><affiliation wicri:level="1"><nlm:affiliation>HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR</wicri:regionArea><wicri:noRegion>Hong Kong SAR</wicri:noRegion></affiliation></author><author><name sortKey="Teoh, K T" sort="Teoh, K T" uniqKey="Teoh K" first="K T" last="Teoh">K T Teoh</name></author><author><name sortKey="Lo, J" sort="Lo, J" uniqKey="Lo J" first="J" last="Lo">J. Lo</name></author><author><name sortKey="Chan, C M" sort="Chan, C M" uniqKey="Chan C" first="C M" last="Chan">C M Chan</name></author><author><name sortKey="Kien, F" sort="Kien, F" uniqKey="Kien F" first="F" last="Kien">F. Kien</name></author><author><name sortKey="Escriou, N" sort="Escriou, N" uniqKey="Escriou N" first="N" last="Escriou">N. Escriou</name></author><author><name sortKey="Tsao, S W" sort="Tsao, S W" uniqKey="Tsao S" first="S W" last="Tsao">S W Tsao</name></author><author><name sortKey="Nicholls, J M" sort="Nicholls, J M" uniqKey="Nicholls J" first="J M" last="Nicholls">J M Nicholls</name></author><author><name sortKey="Altmeyer, R" sort="Altmeyer, R" uniqKey="Altmeyer R" first="R" last="Altmeyer">R. Altmeyer</name></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J S M" last="Peiris">J S M. Peiris</name></author><author><name sortKey="Bruzzone, R" sort="Bruzzone, R" uniqKey="Bruzzone R" first="R" last="Bruzzone">R. Bruzzone</name></author><author><name sortKey="Nal, B" sort="Nal, B" uniqKey="Nal B" first="B" last="Nal">B. Nal</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:18753196</idno><idno type="pmid">18753196</idno><idno type="doi">10.1128/JVI.01052-08</idno><idno type="wicri:Area/PubMed/Corpus">001A77</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001A77</idno><idno type="wicri:Area/PubMed/Curation">001A77</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001A77</idno><idno type="wicri:Area/PubMed/Checkpoint">001997</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001997</idno><idno type="wicri:Area/Ncbi/Merge">001D36</idno><idno type="wicri:Area/Ncbi/Curation">001D36</idno><idno type="wicri:Area/Ncbi/Checkpoint">001D36</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles.</title><author><name sortKey="Siu, Y L" sort="Siu, Y L" uniqKey="Siu Y" first="Y L" last="Siu">Y L Siu</name><affiliation wicri:level="1"><nlm:affiliation>HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR</wicri:regionArea><wicri:noRegion>Hong Kong SAR</wicri:noRegion></affiliation></author><author><name sortKey="Teoh, K T" sort="Teoh, K T" uniqKey="Teoh K" first="K T" last="Teoh">K T Teoh</name></author><author><name sortKey="Lo, J" sort="Lo, J" uniqKey="Lo J" first="J" last="Lo">J. Lo</name></author><author><name sortKey="Chan, C M" sort="Chan, C M" uniqKey="Chan C" first="C M" last="Chan">C M Chan</name></author><author><name sortKey="Kien, F" sort="Kien, F" uniqKey="Kien F" first="F" last="Kien">F. Kien</name></author><author><name sortKey="Escriou, N" sort="Escriou, N" uniqKey="Escriou N" first="N" last="Escriou">N. Escriou</name></author><author><name sortKey="Tsao, S W" sort="Tsao, S W" uniqKey="Tsao S" first="S W" last="Tsao">S W Tsao</name></author><author><name sortKey="Nicholls, J M" sort="Nicholls, J M" uniqKey="Nicholls J" first="J M" last="Nicholls">J M Nicholls</name></author><author><name sortKey="Altmeyer, R" sort="Altmeyer, R" uniqKey="Altmeyer R" first="R" last="Altmeyer">R. Altmeyer</name></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J S M" last="Peiris">J S M. Peiris</name></author><author><name sortKey="Bruzzone, R" sort="Bruzzone, R" uniqKey="Bruzzone R" first="R" last="Bruzzone">R. Bruzzone</name></author><author><name sortKey="Nal, B" sort="Nal, B" uniqKey="Nal B" first="B" last="Nal">B. Nal</name></author></analytic><series><title level="j">Journal of virology</title><idno type="eISSN">1098-5514</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Humans</term><term>Membrane Glycoproteins (metabolism)</term><term>Microscopy, Electron, Transmission</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (metabolism)</term><term>SARS Virus (physiology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (metabolism)</term><term>Viral Matrix Proteins (genetics)</term><term>Viral Matrix Proteins (metabolism)</term><term>Virosomes (metabolism)</term><term>Virosomes (ultrastructure)</term><term>Virus Assembly</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Assemblage viral</term><term>Cellules Vero</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Microscopie électronique à transmission</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines de la matrice virale (métabolisme)</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (métabolisme)</term><term>Virosomes (métabolisme)</term><term>Virosomes (ultrastructure)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Nucleocapsid Proteins</term><term>Viral Envelope Proteins</term><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term><term>Nucleocapsid Proteins</term><term>Viral Envelope Proteins</term><term>Viral Matrix Proteins</term><term>Virosomes</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines de l'enveloppe virale</term><term>Protéines de la matrice virale</term><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines de la matrice virale</term><term>Protéines nucléocapside</term><term>Virosomes</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="ultrastructure" xml:lang="en"><term>Virosomes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Humans</term><term>Microscopy, Electron, Transmission</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Virus Assembly</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="fr"><term>Animaux</term><term>Assemblage viral</term><term>Cellules Vero</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term><term>Microscopie électronique à transmission</term><term>Virosomes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The production of virus-like particles (VLPs) constitutes a relevant and safe model to study molecular determinants of virion egress. The minimal requirement for the assembly of VLPs for the coronavirus responsible for severe acute respiratory syndrome in humans (SARS-CoV) is still controversial. Recent studies have shown that SARS-CoV VLP formation depends on either M and E proteins or M and N proteins. Here we show that both E and N proteins must be coexpressed with M protein for the efficient production and release of VLPs by transfected Vero E6 cells. This suggests that the mechanism of SARS-CoV assembly differs from that of other studied coronaviruses, which only require M and E proteins for VLP formation. When coexpressed, the native envelope trimeric S glycoprotein is incorporated onto VLPs. Interestingly, when a fluorescent protein tag is added to the C-terminal end of N or S protein, but not M protein, the chimeric viral proteins can be assembled within VLPs and allow visualization of VLP production and trafficking in living cells by state-of-the-art imaging technologies. Fluorescent VLPs will be used further to investigate the role of cellular machineries during SARS-CoV egress.</div></front></TEI></PubMed></double></record>Pour manipuler ce document sous Unix (Dilib)
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