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 Yao, J.
Right arrow Articles by Gillam, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yao, J.
Right arrow Articles by Gillam, S.

 Previous Article  |  Next Article 

Journal of Virology, June 1999, p. 4622-4630, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Mutational Analysis, Using a Full-Length Rubella Virus cDNA Clone, of Rubella Virus E1 Transmembrane and Cytoplasmic Domains Required for Virus Release

Jiansheng Yao and Shirley Gillam*

Department of Pathology and Laboratory Medicine, Research Institute, University of British Columbia, Vancouver, British Columbia V5Z 4H4, Canada

Received 10 July 1998/Accepted 23 February 1999

We report on the construction of a full-length cDNA clone, pBRM33, derived from wild-type rubella virus M33 strain. The RNA transcripts synthesized in vitro from pBRM33 are highly infectious, and the viruses produced retain the phenotypic characteristics of the parental M33 virus in growth rate and plaque size. This cDNA clone was used to study the role of E1 transmembrane and cytoplasmic domains in virus assembly by site-directed mutagenesis. Three different alanine substitutions were introduced in the transmembrane domain of E1. These included substitution of leucine 464, cysteine 466, cysteine 467, and both cysteines 466 and 467 to alanine. In the E1 cytoplasmic domain, cysteine 470 and leucine 471 were altered to alanine. We found that these mutations did not significantly affect viral RNA replication, viral structural protein synthesis and transport, or E2/E1 heterodimer formation. Except for the substitution of cysteine 470, these mutations did, however, lead to a reduction in virus release. Substitution of cysteine 467 in the transmembrane region and of leucine 471 in the cytoplasmic domain dramatically reduced virus yield, resulting in the production of only 1 and 10% of the parental virus yield, respectively, in a parallel infection. These data show that E1 transmembrane and cytoplasmic domains play an important role in late stages of virus assembly, possibly during virus budding, consistent with earlier studies indicating that the E1 cytoplasmic domain may interact with nucleocapsids and that this interaction drives virus budding.

* Corresponding author. Mailing address: Department of Pathology and Laboratory Medicine, University of British Columbia, Research Institute, 950 West 28th Ave., Vancouver, British Columbia V5Z 4H4, Canada. Phone: (604) 875-2474. Fax: (604) 875-2496. E-mail: gillam{at}wpog.childhosp.bc.ca.

Journal of Virology, June 1999, p. 4622-4630, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Ilkow, C. S., Mancinelli, V., Beatch, M. D., Hobman, T. C. (2008). Rubella Virus Capsid Protein Interacts with Poly(A)-Binding Protein and Inhibits Translation. J. Virol. 82: 4284-4294 [Abstract] [Full Text]  
  • Overby, A. K., Popov, V. L., Pettersson, R. F., Neve, E. P. A. (2007). The Cytoplasmic Tails of Uukuniemi Virus (Bunyaviridae) GN and GC Glycoproteins Are Important for Intracellular Targeting and the Budding of Virus-Like Particles. J. Virol. 81: 11381-11391 [Abstract] [Full Text]  
  • Law, L. J., Ilkow, C. S., Tzeng, W.-P., Rawluk, M., Stuart, D. T., Frey, T. K., Hobman, T. C. (2006). Analyses of Phosphorylation Events in the Rubella Virus Capsid Protein: Role in Early Replication Events. J. Virol. 80: 6917-6925 [Abstract] [Full Text]  
  • Tzeng, W.-P., Matthews, J. D., Frey, T. K. (2006). Analysis of rubella virus capsid protein-mediated enhancement of replicon replication and mutant rescue.. J. Virol. 80: 3966-3974 [Abstract] [Full Text]  
  • Beatch, M. D., Everitt, J. C., Law, L. J., Hobman, T. C. (2005). Interactions between Rubella Virus Capsid and Host Protein p32 Are Important for Virus Replication. J. Virol. 79: 10807-10820 [Abstract] [Full Text]  
  • Law, L. M. J., Everitt, J. C., Beatch, M. D., Holmes, C. F. B., Hobman, T. C. (2003). Phosphorylation of Rubella Virus Capsid Regulates Its RNA Binding Activity and Virus Replication. J. Virol. 77: 1764-1771 [Abstract] [Full Text]  
  • Lee, J.-Y., Bowden, D. S. (2000). Rubella Virus Replication and Links to Teratogenicity. Clin. Microbiol. Rev. 13: 571-587 [Abstract] [Full Text]  
  • Qiu, Z., Yao, J., Cao, H., Gillam, S. (2000). Mutations in the E1 Hydrophobic Domain of Rubella Virus Impair Virus Infectivity but Not Virus Assembly. J. Virol. 74: 6637-6642 [Abstract] [Full Text]  
  • Liang, Y., Yao, J., Gillam, S. (2000). Rubella Virus Nonstructural Protein Protease Domains Involved in trans- and cis-Cleavage Activities. J. Virol. 74: 5412-5423 [Abstract] [Full Text]  
  • Liang, Y., Gillam, S. (2000). Mutational Analysis of the Rubella Virus Nonstructural Polyprotein and Its Cleavage Products in Virus Replication and RNA Synthesis. J. Virol. 74: 5133-5141 [Abstract] [Full Text]  
  • Yao, J., Gillam, S. (2000). A Single-Amino-Acid Substitution of a Tyrosine Residue in the Rubella Virus E1 Cytoplasmic Domain Blocks Virus Release. J. Virol. 74: 3029-3036 [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»