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, November 2002, p. 10829-10840, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10829-10840.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of Two New Structural Glycoproteins, GP3 and GP4, of Equine Arteritis Virus

Roeland Wieringa,* Antoine A. F. de Vries,{dagger} Martin J. B. Raamsman,2 and Peter J. M. Rottier

Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, and Institute of Biomembranes, Utrecht University, 3584 CL Utrecht, The Netherlands

Received 3 April 2002/ Accepted 22 July 2002

Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. Four envelope proteins have hitherto been identified in EAV particles: the predominant membrane proteins M and GL, the unglycosylated small envelope protein E, and the nonabundant membrane glycoprotein GS. In this study, we established that the products of EAV open reading frame 3 (ORF3) and ORF4 (designated GP3 and GP4, respectively) are also minor structural glycoproteins. The proteins were first characterized by various analyses after in vitro translation of RNA transcripts in a rabbit reticulocyte lysate in the presence and absence of microsomal membranes. We subsequently expressed ORF3 and -4 in baby hamster kidney cells by using the vaccinia virus expression system and, finally, analyzed the GP3 and GP4 proteins synthesized in EAV-infected cells. The results showed that GP4 is a class I integral membrane protein of 28 kDa with three functional N-glycosylation sites and with little, if any, of its carboxy terminus exposed. Both after independent expression and in EAV-infected cells, the protein localizes in the endoplasmic reticulum (ER), as demonstrated biochemically by analysis of its oligosaccharide side chains and as visualized directly by immunofluorescence studies. GP3, on the other hand, is a heavily glycosylated protein whose hydrophobic amino terminus is not cleaved off. It is an integral membrane protein anchored by either or both of its hydrophobic terminal domains and with no parts detectably exposed cytoplasmically. Also, GP3 localizes in the ER when expressed independently and in the context of an EAV infection. Only a small fraction of the GP3 and GP4 proteins synthesized in infected cells ends up in virions. Most, but not all, of the oligosaccharides of these virion glycoproteins are biochemically mature. Our results bring the number of EAV envelope proteins to six.

* 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: r.wieringa{at}vet.uu.nl.

{dagger} Present address: Gene Therapy Section, Department of Molecular Cell Biology, Leiden University Medical Center (LUMC), 2333 AL Leiden, The Netherlands.

{ddagger} Present address: Crucell N.V., 2333 CN Leiden, The Netherlands.

Journal of Virology, November 2002, p. 10829-10840, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10829-10840.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thaa, B., Kabatek, A., Zevenhoven-Dobbe, J. C., Snijder, E. J., Herrmann, A., Veit, M. (2009). Myristoylation of the arterivirus E protein: the fatty acid modification is not essential for membrane association but contributes significantly to virus infectivity. J. Gen. Virol. 90: 2704-2712 [Abstract] [Full Text]  
  • Go, Y. Y., Wong, S. J., Branscum, A. J., Demarest, V. L., Shuck, K. M., Vickers, M. L., Zhang, J., McCollum, W. H., Timoney, P. J., Balasuriya, U. B. R. (2008). Development of a Fluorescent-Microsphere Immunoassay for Detection of Antibodies Specific to Equine Arteritis Virus and Comparison with the Virus Neutralization Test. CVI 15: 76-87 [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]  
  • Wieringa, R., de Vries, A. A. F., van der Meulen, J., Godeke, G.-J., Onderwater, J. J. M., van Tol, H., Koerten, H. K., Mommaas, A. M., Snijder, E. J., Rottier, P. J. M. (2004). Structural Protein Requirements in Equine Arteritis Virus Assembly. J. Virol. 78: 13019-13027 [Abstract] [Full Text]  
  • Wieringa, R., de Vries, A. A. F., Post, S. M., Rottier, P. J. M. (2003). Intra- and Intermolecular Disulfide Bonds of the GP2b Glycoprotein of Equine Arteritis Virus: Relevance for Virus Assembly and Infectivity. J. Virol. 77: 12996-13004 [Abstract] [Full Text]  
  • Castillo-Olivares, J., Wieringa, R., Bakonyi, T., de Vries, A. A. F., Davis-Poynter, N. J., Rottier, P. J. M. (2003). Generation of a Candidate Live Marker Vaccine for Equine Arteritis Virus by Deletion of the Major Virus Neutralization Domain. J. Virol. 77: 8470-8480 [Abstract] [Full Text]  
  • Wieringa, R., de Vries, A. A. F., Rottier, P. J. M. (2003). Formation of Disulfide-Linked Complexes between the Three Minor Envelope Glycoproteins (GP2b, GP3, and GP4) of Equine Arteritis Virus. J. Virol. 77: 6216-6226 [Abstract] [Full Text]  
  • Snijder, E. J., Dobbe, J. C., Spaan, W. J. M. (2003). Heterodimerization of the Two Major Envelope Proteins Is Essential for Arterivirus Infectivity. J. Virol. 77: 97-104 [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»