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 Hassan, S. H.
Right arrow Articles by Roy, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hassan, S. H.
Right arrow Articles by Roy, P.

 Previous Article  |  Next Article 

Journal of Virology, September 2001, p. 8356-8367, Vol. 75, No. 18
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.18.8356-8367.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Expression and Functional Characterization of Bluetongue Virus VP5 Protein: Role in Cellular Permeabilization

S. H. Hassan,1 C. Wirblich,1 M. Forzan,1 and P. Roy1,2,*

Department of Infectious and Tropical Diseases, School of Hygiene and Tropical Medicine, London WC1E 7HT, England,1 and Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 352942

Received 28 February 2001/Accepted 8 June 2001

Segment 5 of bluetongue virus (BTV) serotype 10, which encodes the outer capsid protein VP5, was tagged with glutathione S-transferase and expressed by a recombinant baculovirus. The recombinant protein was subsequently purified to homogeneity, and its possible biological role in virus infection was investigated. Purified VP5 was able to bind mammalian cells but was not internalized, which indicates it is not involved in receptor-mediated endocytosis. The purified VP5 protein was shown to be able to permeabilize mammalian and Culicoides insect cells, inducing cytotoxicity. Sequence analysis revealed that VP5 possesses characteristic structural features (including two amino-terminal amphipathic helices) compatible with virus penetration activity. To assess the role of each feature in the observed cytotoxicity, a series of deleted VP5 molecules were generated, and their expression and biological activity was compared with the parental molecule. VP5 derivatives that included the two amphipathic helices exhibited cytotoxicity, while those that omitted these sequences did not. To confirm their role in membrane destabilization two synthetic peptides (amino acids [aa] 1 to 20 and aa 22 to 41) encompassing the two helices and an additional peptide representing the adjacent downstream sequences were also assessed for their effect on the cell membrane. Both helices, but not the downstream VP5 sequence, exhibited cytotoxicity with the most-amino-terminal helix (aa 1 to 20) showing a higher activity than the adjacent peptide (aa 22 to 41). Purified VP5 was shown to readily form trimers in solution, a feature of many proteins involved in membrane penetration. Taken together, these data support a role for VP5 in virus-cell penetration consistent with its revelation in the entry vesicle subsequent to cell binding and endocytosis.

* Corresponding author. Mailing address: Department of Infectious and Tropical Diseases, School of Hygiene and Tropical Medicine, Keppel St., London WC1E 7HT, England. Phone: 44 (0) 20-7927-2324. Fax: 44 (0) 20-7636-8739. E-mail: polly.roy{at}lshtm.ac.uk.

Journal of Virology, September 2001, p. 8356-8367, Vol. 75, No. 18
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.18.8356-8367.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Bhattacharya, B., Roy, P. (2008). Bluetongue Virus Outer Capsid Protein VP5 Interacts with Membrane Lipid Rafts via a SNARE Domain. J. Virol. 82: 10600-10612 [Abstract] [Full Text]  
  • Roy, P. (2008). Bluetongue virus: dissection of the polymerase complex. J. Gen. Virol. 89: 1789-1804 [Abstract] [Full Text]  
  • Forzan, M., Marsh, M., Roy, P. (2007). Bluetongue Virus Entry into Cells. J. Virol. 81: 4819-4827 [Abstract] [Full Text]  
  • Modrof, J., Lymperopoulos, K., Roy, P. (2005). Phosphorylation of Bluetongue Virus Nonstructural Protein 2 Is Essential for Formation of Viral Inclusion Bodies. J. Virol. 79: 10023-10031 [Abstract] [Full Text]  
  • Nason, E. L., Rothagel, R., Mukherjee, S. K., Kar, A. K., Forzan, M., Prasad, B. V. V., Roy, P. (2004). Interactions between the Inner and Outer Capsids of Bluetongue Virus. J. Virol. 78: 8059-8067 [Abstract] [Full Text]  
  • Mortola, E., Noad, R., Roy, P. (2004). Bluetongue Virus Outer Capsid Proteins Are Sufficient To Trigger Apoptosis in Mammalian Cells. J. Virol. 78: 2875-2883 [Abstract] [Full Text]  
  • Forzan, M., Wirblich, C., Roy, P. (2004). A capsid protein of nonenveloped Bluetongue virus exhibits membrane fusion activity. Proc. Natl. Acad. Sci. USA 101: 2100-2105 [Abstract] [Full Text]  
  • Beaton, A. R., Rodriguez, J., Reddy, Y. K., Roy, P. (2002). The membrane trafficking protein calpactin forms a complex with bluetongue virus protein NS3 and mediates virus release. Proc. Natl. Acad. Sci. USA 99: 13154-13159 [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»