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Article: 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

TitleThe 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
Authors
Issue Date2008
PublisherAmerican Society for Microbiology. The Journal's web site is located at http://jvi.asm.org/
Citation
Journal Of Virology, 2008, v. 82 n. 22, p. 11318-11330 How to Cite?
AbstractThe 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. Copyright © 2008, American Society for Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/132773
ISSN
2015 Impact Factor: 4.606
2015 SCImago Journal Rankings: 3.347
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
University of Hong Kong
French Ministry of Health
French Chancery of Paris Universities
EU-6th Framework Program (EPISARS)
Funding Information:

We thank K. H. Chan (Department of Microbiology, University of Hong Kong) for the gift of the mouse monoclonal antibody against the SARS-CoV N protein, V. Lorin (Institut Pasteur) for preparation and analysis of the anti- E rabbit serum, and Roger Y. Tsien (University of California, San Diego) for providing the plasmid coding for the mRFP1 protein. We especially thank the Electron Microscope Unit of the University of Hong Kong, Li Ka Shing Faculty of Medicine, and Marie-Christine Prevost and Martin Sachse (Plate-Forme de Microscopie Electronique, Institut Pasteur) for expert advice on the electron microscopy experiments; Iris Ng (Department of Microbiology, University of Hong Kong) for technical support for electron microscopy experiments; and Tony Chan (Department of Anatomy, Li Ka Shing Faculty of Medicine Core Imaging Facility, University of Hong Kong) for technical support during the live cell imaging experiments.

References

 

DC FieldValueLanguage
dc.contributor.authorSiu, YLen_HK
dc.contributor.authorTeoh, KTen_HK
dc.contributor.authorLo, Jen_HK
dc.contributor.authorChan, CMen_HK
dc.contributor.authorKien, Fen_HK
dc.contributor.authorEscriou, Nen_HK
dc.contributor.authorTsao, SWen_HK
dc.contributor.authorNicholls, JMen_HK
dc.contributor.authorAltmeyer, Ren_HK
dc.contributor.authorPeiris, JSMen_HK
dc.contributor.authorBruzzone, Ren_HK
dc.contributor.authorNal, Ben_HK
dc.date.accessioned2011-03-28T09:28:53Z-
dc.date.available2011-03-28T09:28:53Z-
dc.date.issued2008en_HK
dc.identifier.citationJournal Of Virology, 2008, v. 82 n. 22, p. 11318-11330en_HK
dc.identifier.issn0022-538Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/132773-
dc.description.abstractThe 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. Copyright © 2008, American Society for Microbiology. All Rights Reserved.en_HK
dc.languageengen_US
dc.publisherAmerican Society for Microbiology. The Journal's web site is located at http://jvi.asm.org/en_HK
dc.relation.ispartofJournal of Virologyen_HK
dc.rightsJournal of Virology. Copyright © American Society for Microbiology.-
dc.rightsCopyright © American Society for Microbiology, Journal of Virology, 2008, v. 82 n. 22, p. 11318-11330-
dc.subject.meshNucleocapsid Proteins - genetics - metabolism-
dc.subject.meshSARS Virus - physiology-
dc.subject.meshViral Envelope Proteins - genetics - metabolism-
dc.subject.meshViral Matrix Proteins - genetics - metabolism-
dc.subject.meshVirus Assembly-
dc.titleThe M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particlesen_HK
dc.typeArticleen_HK
dc.identifier.emailTsao, SW: gswtsao@hku.hken_HK
dc.identifier.emailNicholls, JM: jmnichol@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_HK
dc.identifier.emailBruzzone, R: bruzzone@hkucc.hku.hken_HK
dc.identifier.emailNal, B: bnal@hkucc.hku.hken_HK
dc.identifier.authorityTsao, SW=rp00399en_HK
dc.identifier.authorityNicholls, JM=rp00364en_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.identifier.authorityBruzzone, R=rp01442en_HK
dc.identifier.authorityNal, B=rp00541en_HK
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1128/JVI.01052-08en_HK
dc.identifier.pmid18753196-
dc.identifier.pmcidPMC2573274-
dc.identifier.scopuseid_2-s2.0-55549119619en_HK
dc.identifier.hkuros162673-
dc.identifier.hkuros149936-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-55549119619&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume82en_HK
dc.identifier.issue22en_HK
dc.identifier.spage11318en_HK
dc.identifier.epage11330en_HK
dc.identifier.isiWOS:000260368000032-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridSiu, YL=25227033200en_HK
dc.identifier.scopusauthoridTeoh, KT=25637993600en_HK
dc.identifier.scopusauthoridLo, J=35748856900en_HK
dc.identifier.scopusauthoridChan, CM=7404814453en_HK
dc.identifier.scopusauthoridKien, F=7004231633en_HK
dc.identifier.scopusauthoridEscriou, N=6603606703en_HK
dc.identifier.scopusauthoridTsao, SW=7102813116en_HK
dc.identifier.scopusauthoridNicholls, JM=7201463077en_HK
dc.identifier.scopusauthoridAltmeyer, R=7003677186en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.scopusauthoridBruzzone, R=7006793327en_HK
dc.identifier.scopusauthoridNal, B=6506672380en_HK

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