This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wieringa, R.
Right arrow Articles by Rottier, P. J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wieringa, R.
Right arrow Articles by Rottier, P. J. M.

 Previous Article  |  Next Article 

Journal of Virology, December 2004, p. 13019-13027, Vol. 78, No. 23
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.23.13019-13027.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Structural Protein Requirements in Equine Arteritis Virus Assembly

Roeland Wieringa,1,{dagger} Antoine A. F. de Vries,2 Jannes van der Meulen,3 Gert-Jan Godeke,1,{ddagger} Jos J. M. Onderwater,3 Hans van Tol,4 Henk K. Koerten,3 A. Mieke Mommaas,3 Eric J. Snijder,4 and Peter J. M. Rottier1*

Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, and Institute of Biomembranes, Utrecht University, Utrecht,1 Gene Therapy Section,2 Center for Electron Microscopy, Department of Molecular Cell Biology,3 Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands4

Received 4 May 2004/ Accepted 20 July 2004

Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. EAV particles contain seven structural proteins: the nucleocapsid protein N, the unglycosylated envelope proteins M and E, and the N-glycosylated membrane proteins GP2b (previously named GS), GP3, GP4, and GP5 (previously named GL). Proteins N, M, and GP5 are major virion components, E occurs in virus particles in intermediate amounts, and GP4, GP3, and GP2b are minor structural proteins. The M and GP5 proteins occur in virus particles as disulfide-linked heterodimers while the GP4, GP3, and GP2b proteins are incorporated into virions as a heterotrimeric complex. Here, we studied the effect on virus assembly of inactivating the structural protein genes one by one in the context of a (full-length) EAV cDNA clone. It appeared that the three major structural proteins are essential for particle formation, while the other four virion proteins are dispensable. When one of the GP2b, GP3, or GP4 proteins was missing, the incorporation of the remaining two minor envelope glycoproteins was completely blocked while that of the E protein was greatly reduced. The absence of E entirely prevented the incorporation of the GP2b, GP3, and GP4 proteins into viral particles. EAV particles lacking GP2b, GP3, GP4, and E did not markedly differ from wild-type virions in buoyant density, major structural protein composition, electron microscopic appearance, and genomic RNA content. On the basis of these results, we propose a model for the EAV particle in which the GP2b/GP3/GP4 heterotrimers are positioned, in association with a defined number of E molecules, above the vertices of the putatively icosahedral nucleocapsid.

* Corresponding author. Mailing address: Virology Division, Department of Infectious Diseases and Immunology, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Phone: 31 30 2532463. Fax: 31 30 2536723. E-mail: p.rottier{at}vet.uu.nl.

{dagger} Present address: Gene Therapy Section, Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.

{ddagger} Present address: Microbiological Laboratory for Health Protection, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.

Journal of Virology, December 2004, p. 13019-13027, Vol. 78, No. 23
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.23.13019-13027.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tijms, M. A., Nedialkova, D. D., Zevenhoven-Dobbe, J. C., Gorbalenya, A. E., Snijder, E. J. (2007). Arterivirus Subgenomic mRNA Synthesis and Virion Biogenesis Depend on the Multifunctional nsp1 Autoprotease. J. Virol. 81: 10496-10505 [Abstract] [Full Text]  
  • van den Born, E., Posthuma, C. C., Knoops, K., Snijder, E. J. (2007). An infectious recombinant equine arteritis virus expressing green fluorescent protein from its replicase gene. J. Gen. Virol. 88: 1196-1205 [Abstract] [Full Text]  
  • Balasuriya, U. B. R., Snijder, E. J., Heidner, H. W., Zhang, J., Zevenhoven-Dobbe, J. C., Boone, J. D., McCollum, W. H., Timoney, P. J., MacLachlan, N. J. (2007). Development and characterization of an infectious cDNA clone of the virulent Bucyrus strain of Equine arteritis virus. J. Gen. Virol. 88: 918-924 [Abstract] [Full Text]  
  • Costers, S., Delputte, P. L., Nauwynck, H. J. (2006). Porcine reproductive and respiratory syndrome virus-infected alveolar macrophages contain no detectable levels of viral proteins in their plasma membrane and are protected against antibody-dependent, complement-mediated cell lysis.. J. Gen. Virol. 87: 2341-2351 [Abstract] [Full Text]  
  • Neuman, B. W., Adair, B. D., Yoshioka, C., Quispe, J. D., Orca, G., Kuhn, P., Milligan, R. A., Yeager, M., Buchmeier, M. J. (2006). Supramolecular architecture of severe acute respiratory syndrome coronavirus revealed by electron cryomicroscopy.. J. Virol. 80: 7918-7928 [Abstract] [Full Text]  
  • Oostra, M., de Haan, C. A. M., de Groot, R. J., Rottier, P. J. M. (2006). Glycosylation of the Severe Acute Respiratory Syndrome Coronavirus Triple-Spanning Membrane Proteins 3a and M. J. Virol. 80: 2326-2336 [Abstract] [Full Text]  
  • Posthuma, C. C., Nedialkova, D. D., Zevenhoven-Dobbe, J. C., Blokhuis, J. H., Gorbalenya, A. E., Snijder, E. J. (2006). Site-Directed Mutagenesis of the Nidovirus Replicative Endoribonuclease NendoU Exerts Pleiotropic Effects on the Arterivirus Life Cycle. J. Virol. 80: 1653-1661 [Abstract] [Full Text]  
  • Choi, Y.-J., Yun, S.-I., Kang, S.-Y., Lee, Y.-M. (2006). Identification of 5' and 3' cis-Acting Elements of the Porcine Reproductive and Respiratory Syndrome Virus: Acquisition of Novel 5' AU-Rich Sequences Restored Replication of a 5'-Proximal 7-Nucleotide Deletion Mutant. J. Virol. 80: 723-736 [Abstract] [Full Text]  
  • Yount, B., Roberts, R. S., Sims, A. C., Deming, D., Frieman, M. B., Sparks, J., Denison, M. R., Davis, N., Baric, R. S. (2005). Severe Acute Respiratory Syndrome Coronavirus Group-Specific Open Reading Frames Encode Nonessential Functions for Replication in Cell Cultures and Mice. J. Virol. 79: 14909-14922 [Abstract] [Full Text]  
  • Lee, C., Yoo, D. (2005). Cysteine residues of the porcine reproductive and respiratory syndrome virus small envelope protein are non-essential for virus infectivity. J. Gen. Virol. 86: 3091-3096 [Abstract] [Full Text]  
  • Wissink, E. H. J., Kroese, M. V., van Wijk, H. A. R., Rijsewijk, F. A. M., Meulenberg, J. J. M., Rottier, P. J. M. (2005). Envelope Protein Requirements for the Assembly of Infectious Virions of Porcine Reproductive and Respiratory Syndrome Virus. J. Virol. 79: 12495-12506 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»