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 Katz, E.
Right arrow Articles by Moss, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Katz, E.
Right arrow Articles by Moss, B.

 Previous Article  |  Next Article 

Journal of Virology, November 2002, p. 11637-11644, Vol. 76, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.22.11637-11644.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of Second-Site Mutations That Enhance Release and Spread of Vaccinia Virus

Ehud Katz,{dagger} Elizabeth Wolffe,{ddagger} and Bernard Moss*

Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0445

Received 10 July 2002/ Accepted 19 August 2002

The spread of most strains of vaccinia virus in cell monolayers occurs predominantly via extracellular enveloped virions that adhere to the tips of actin-containing microvilli and to a lesser extent via diffusion of released virions. The mechanism by which virions adhere to the cell surface is unknown, although several viral proteins may be involved. The present investigation was initiated with the following premise: spontaneous mutations that increase virus release will be naturally selected by propagating a virus unable to spread by means of actin tails. Starting with an A36R deletion mutant that forms small, round plaques, five independent virus clones with enhanced spread due to the formation of comet or satellite plaques were isolated. The viral membrane glycoprotein genes of the isolates were sequenced; four had mutations causing C-terminal truncations of the A33R protein, and one had a serine replacing proline 189 of the B5R protein. The comet-forming phenotype was specifically reproduced or reversed by homologous recombination using DNA containing the mutated or natural sequence, respectively. Considerably more extracellular enveloped virus was released into the medium by the second-site mutants than by the parental A36R deletion mutant, explaining their selection in tissue culture as well as their comet-forming phenotype. The data suggest that the B5R protein and the C-terminal region of the A33R protein are involved in adherence of cell-associated enveloped virions to cells. In spite of their selective advantage in cultured cells, the second-site mutants were not detectably more virulent than the A36R deletion mutant when administered to mice by the intranasal route.

* Corresponding author. Mailing address: 4 Center Dr., National Institutes of Health, Bethesda, MD 20892-0445. Phone: (301) 496-9869. Fax: (301) 480-1147. E-mail: bmoss{at}nih.gov.

{dagger} Permanent address: Department of Virology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.

{ddagger} Present address: Sangamo BioSciences, Inc., Richmond, CA 94804.

Journal of Virology, November 2002, p. 11637-11644, Vol. 76, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.22.11637-11644.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Olson, V. A., Karem, K. L., Smith, S. K., Hughes, C. M., Damon, I. K. (2009). Smallpox virus plaque phenotypes: genetic, geographical and case fatality relationships. J. Gen. Virol. 90: 792-798 [Abstract] [Full Text]  
  • Berhanu, A., Wilson, R. L., Kirkwood-Watts, D. L., King, D. S., Warren, T. K., Lund, S. A., Brown, L. L., Krupkin, A. K., VanderMay, E., Weimers, W., Honeychurch, K. M., Grosenbach, D. W., Jones, K. F., Hruby, D. E. (2008). Vaccination of BALB/c Mice with Escherichia coli-Expressed Vaccinia Virus Proteins A27L, B5R, and D8L Protects Mice from Lethal Vaccinia Virus Challenge. J. Virol. 82: 3517-3529 [Abstract] [Full Text]  
  • Perdiguero, B., Lorenzo, M. M., Blasco, R. (2008). Vaccinia Virus A34 Glycoprotein Determines the Protein Composition of the Extracellular Virus Envelope. J. Virol. 82: 2150-2160 [Abstract] [Full Text]  
  • Perdiguero, B., Blasco, R. (2006). Interaction between Vaccinia Virus Extracellular Virus Envelope A33 and B5 Glycoproteins.. J. Virol. 80: 8763-8777 [Abstract] [Full Text]  
  • Chan, K. S., Verardi, P. H., Legrand, F. A., Yilma, T. D. (2005). Nef from pathogenic simian immunodeficiency virus is a negative factor for vaccinia virus. Proc. Natl. Acad. Sci. USA 102: 8734-8739 [Abstract] [Full Text]  
  • Tenorio, A. R., Peeples, M. E., Patri, M., Rezai, K., Trenholme, G. M. (2004). Evolution of Vaccinia Virus-Specific CD8+ Cytotoxic T-Lymphocyte Responses in Primary Vaccinees and Revaccinees. CVI 11: 758-761 [Abstract] [Full Text]  
  • Katz, E., Ward, B. M., Weisberg, A. S., Moss, B. (2003). Mutations in the Vaccinia Virus A33R and B5R Envelope Proteins That Enhance Release of Extracellular Virions and Eliminate Formation of Actin-Containing Microvilli without Preventing Tyrosine Phosphorylation of the A36R Protein. J. Virol. 77: 12266-12275 [Abstract] [Full Text]  
  • Carter, G. C., Rodger, G., Murphy, B. J., Law, M., Krauss, O., Hollinshead, M., Smith, G. L. (2003). Vaccinia virus cores are transported on microtubules. J. Gen. Virol. 84: 2443-2458 [Abstract] [Full Text]  
  • Meiser, A., Boulanger, D., Sutter, G., Krijnse Locker, J. (2003). Comparison of virus production in chicken embryo fibroblasts infected with the WR, IHD-J and MVA strains of vaccinia virus: IHD-J is most efficient in trans-Golgi network wrapping and extracellular enveloped virus release. J. Gen. Virol. 84: 1383-1392 [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»