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Journal of Virology, November 2008, p. 11318-11330, Vol. 82, No. 22
0022-538X/08/$08.00+0     doi:10.1128/JVI.01052-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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 {triangledown} ,{dagger}

Y. L. Siu,1 K. T. Teoh,1 J. Lo,2 C. M. Chan,3 F. Kien,1 N. Escriou,4 S. W. Tsao,5 J. M. Nicholls,2 R. Altmeyer,6 J. S. M. Peiris,1,3 R. Bruzzone,1 and B. Nal1*

HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR, China,1 Department of Pathology, The University of Hong Kong, Queen Mary Hospital, Hong Kong SAR, China,2 Department of Microbiology, The University of Hong Kong, 21 Sassoon Road, Hong Kong SAR, China,3 Institut Pasteur, Unité de Génétique Moléculaire des Virus Respiratoires, URA-CNRS 3015, 25-28 Rue du Docteur Roux, 75724 Paris Cedex 15, France,4 Department of Anatomy, The University of Hong Kong, 21 Sassoon Road, Hong Kong SAR, China,5 CombinatoRx-Singapore Pte, Ltd., 11 Biopolis Way, 138667 Singapore6

Received 20 May 2008/ Accepted 7 August 2008

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.

* Corresponding author. Mailing address: HKU-Pasteur Research Centre, 8 Sassoon Road, Hong Kong SAR, China. Phone: (852) 2816-8403. Fax: (852) 2872-5782. E-mail: bnal{at}hku.hk

{triangledown} Published ahead of print on 27 August 2008.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, November 2008, p. 11318-11330, Vol. 82, No. 22
0022-538X/08/$08.00+0     doi:10.1128/JVI.01052-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Siu, K.-L., Kok, K.-H., Ng, M.-H. J., Poon, V. K. M., Yuen, K.-Y., Zheng, B.-J., Jin, D.-Y. (2009). Severe Acute Respiratory Syndrome Coronavirus M Protein Inhibits Type I Interferon Production by Impeding the Formation of TRAF3{middle dot}TANK{middle dot}TBK1/IKK{epsilon} Complex. J. Biol. Chem. 284: 16202-16209 [Abstract] [Full Text]  


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