1887

Abstract

The dengue virus envelope glycoprotein mediates virus attachment and entry and is the major viral antigen. The identification of ‘critical’ amino acids in the envelope glycoprotein that cannot be altered without loss of infectivity could have a major impact on the development of dengue virus vaccines and diagnostics. In this context, we determined whether six amino acids, previously predicted by computational analysis to play a critical role in the virus life cycle, were essential for virus viability. The effects of mutating the six ‘critical’ amino acids and a further seven ‘neutral’ amino acids were analysed by using a dengue virus type 2 infectious cDNA clone. Of the six critical amino acids, three (Asp-215, Pro-217 and His-244) were found to be essential for virus viability in mammalian and mosquito cells.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.011486-0
2009-10-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/10/2457.html?itemId=/content/journal/jgv/10.1099/vir.0.011486-0&mimeType=html&fmt=ahah

References

  1. Bressanelli, S., Stiasny, K., Allison, S. L., Stura, E. A., Duquerroy, S., Lescar, J., Heinz, F. X. & Rey, F. A. ( 2004; ). Structure of a flavivirus envelope glycoprotein in its low-pH-induced membrane fusion conformation. EMBO J 23, 728–738.[CrossRef]
    [Google Scholar]
  2. Churdboonchart, V., Bhamarapravati, N., Peampramprecha, S. & Sirinavin, S. ( 1991; ). Antibodies against dengue viral proteins in primary and secondary dengue hemorrhagic fever. Am J Trop Med Hyg 44, 481–493.
    [Google Scholar]
  3. Cologna, R. & Rico-Hesse, R. ( 2003; ). American genotype structures decrease dengue virus output from human monocytes and dendritic cells. J Virol 77, 3929–3938.[CrossRef]
    [Google Scholar]
  4. Crill, W. D. & Roehrig, J. T. ( 2001; ). Monoclonal antibodies that bind to domain III of dengue virus E glycoprotein are the most efficient blockers of virus adsorption to Vero cells. J Virol 75, 7769–7773.[CrossRef]
    [Google Scholar]
  5. Fritz, R., Stiasny, K. & Heinz, F. X. ( 2008; ). Identification of specific histidines as pH sensors in flavivirus membrane fusion. J Cell Biol 183, 353–361.[CrossRef]
    [Google Scholar]
  6. Gualano, R. C., Pryor, M. J., Cauchi, M. R., Wright, P. J. & Davidson, A. D. ( 1998; ). Identification of a major determinant of mouse neurovirulence of dengue virus type 2 using stably cloned genomic-length cDNA. J Gen Virol 79, 437–446.
    [Google Scholar]
  7. Gubler, D. J. ( 2002; ). Epidemic dengue/dengue hemorrhagic fever as a public health, social and economic problem in the 21st century. Trends Microbiol 10, 100–103.[CrossRef]
    [Google Scholar]
  8. Hanna, S. L., Pierson, T. C., Sanchez, M. D., Ahmed, A. A., Murtadha, M. M. & Doms, R. W. ( 2005; ). N-linked glycosylation of West Nile virus envelope proteins influences particle assembly and infectivity. J Virol 79, 13262–13274.[CrossRef]
    [Google Scholar]
  9. Kroschewski, H., Lim, S. P., Butcher, R. E., Yap, T. L., Lescar, J., Wright, P. J., Vasudevan, S. G. & Davidson, A. D. ( 2008; ). Mutagenesis of the dengue virus type 2 NS5 methyltransferase domain. J Biol Chem 283, 19410–19421.[CrossRef]
    [Google Scholar]
  10. Kuhn, R. J., Zhang, W., Rossmann, M. G., Pletnev, S. V., Corver, J., Lenches, E., Jones, C. T., Mukhopadhyay, S., Chipman, P. R. & other authors ( 2002; ). Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell 108, 717–725.[CrossRef]
    [Google Scholar]
  11. Kyle, J. L. & Harris, E. ( 2008; ). Global spread and persistence of dengue. Annu Rev Microbiol 62, 71–92.[CrossRef]
    [Google Scholar]
  12. Li, L., Lok, S. M., Yu, I. M., Zhang, Y., Kuhn, R. J., Chen, J. & Rossmann, M. G. ( 2008; ). The flavivirus precursor membrane–envelope protein complex: structure and maturation. Science 319, 1830–1834.[CrossRef]
    [Google Scholar]
  13. Lindenbach, B. D., Thiel, H. J. & Rice, C. M. ( 2007; ). Flaviviridae: the viruses and their replication. In Fields Virology, 5th edn, pp. 1101–1152. Edited by D. M. Knipe & P. M. Howley. Philadelphia, PA: Lippincott Williams & Wilkins.
  14. Mackenzie, J. S., Gubler, D. J. & Petersen, L. R. ( 2004; ). Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nat Med 10, S98–S109.[CrossRef]
    [Google Scholar]
  15. Matusan, A. E., Pryor, M. J., Davidson, A. D. & Wright, P. J. ( 2001; ). Mutagenesis of the dengue virus type 2 NS3 protein within and outside helicase motifs: effects on enzyme activity and virus replication. J Virol 75, 9633–9643.[CrossRef]
    [Google Scholar]
  16. Mazumder, R., Hu, Z. Z., Vinayaka, C. R., Sagripanti, J. L., Frost, S. D., Kosakovsky Pond, S. L. & Wu, C. H. ( 2007; ). Computational analysis and identification of amino acid sites in dengue E proteins relevant to development of diagnostics and vaccines. Virus Genes 35, 175–186.[CrossRef]
    [Google Scholar]
  17. Modis, Y., Ogata, S., Clements, D. & Harrison, S. C. ( 2003; ). A ligand-binding pocket in the dengue virus envelope glycoprotein. Proc Natl Acad Sci U S A 100, 6986–6991.[CrossRef]
    [Google Scholar]
  18. Modis, Y., Ogata, S., Clements, D. & Harrison, S. C. ( 2004; ). Structure of the dengue virus envelope protein after membrane fusion. Nature 427, 313–319.[CrossRef]
    [Google Scholar]
  19. Modis, Y., Ogata, S., Clements, D. & Harrison, S. C. ( 2005; ). Variable surface epitopes in the crystal structure of dengue virus type 3 envelope glycoprotein. J Virol 79, 1223–1231.[CrossRef]
    [Google Scholar]
  20. Mondotte, J. A., Lozach, P. Y., Amara, A. & Gamarnik, A. V. ( 2007; ). Essential role of dengue virus envelope protein N glycosylation at asparagine-67 during viral propagation. J Virol 81, 7136–7148.[CrossRef]
    [Google Scholar]
  21. Mukhopadhyay, S., Kuhn, R. J. & Rossmann, M. G. ( 2005; ). A structural perspective of the flavivirus life cycle. Nat Rev Microbiol 3, 13–22.[CrossRef]
    [Google Scholar]
  22. Pryor, M. J., Carr, J. M., Hocking, H., Davidson, A. D., Li, P. & Wright, P. J. ( 2001; ). Replication of dengue virus type 2 in human monocyte-derived macrophages: comparisons of isolates and recombinant viruses with substitutions at amino acid 390 in the envelope glycoprotein. Am J Trop Med Hyg 65, 427–434.
    [Google Scholar]
  23. Rey, F. A., Heinz, F. X., Mandl, C., Kunz, C. & Harrison, S. C. ( 1995; ). The envelope glycoprotein from tick-borne encephalitis virus at 2 Å resolution. Nature 375, 291–298.[CrossRef]
    [Google Scholar]
  24. Roehrig, J. T. ( 1997; ). Immunochemistry of dengue viruses. In Dengue and Dengue Hemorrhagic Fever, pp. 199–219. Edited by D. Gubler & G. Kuno. New York: CAB International.
  25. Stiasny, K. & Heinz, F. X. ( 2006; ). Flavivirus membrane fusion. J Gen Virol 87, 2755–2766.[CrossRef]
    [Google Scholar]
  26. Wang, W. K., Sung, T. L., Tsai, Y. C., Kao, C. L., Chang, S. M. & King, C. C. ( 2002; ). Detection of dengue virus replication in peripheral blood mononuclear cells from dengue virus type 2-infected patients by a reverse transcription-real-time PCR assay. J Clin Microbiol 40, 4472–4478.[CrossRef]
    [Google Scholar]
  27. Yu, I. M., Zhang, W., Holdaway, H. A., Li, L., Kostyuchenko, V. A., Chipman, P. R., Kuhn, R. J., Rossmann, M. G. & Chen, J. ( 2008; ). Structure of the immature dengue virus at low pH primes proteolytic maturation. Science 319, 1834–1837.[CrossRef]
    [Google Scholar]
  28. Zhang, Y., Corver, J., Chipman, P. R., Zhang, W., Pletnev, S. V., Sedlak, D., Baker, T. S., Strauss, J. H., Kuhn, R. J. & Rossmann, M. G. ( 2003; ). Structures of immature flavivirus particles. EMBO J 22, 2604–2613.[CrossRef]
    [Google Scholar]
  29. Zhang, Y., Zhang, W., Ogata, S., Clements, D., Strauss, J. H., Baker, T. S., Kuhn, R. J. & Rossmann, M. G. ( 2004; ). Conformational changes of the flavivirus E glycoprotein. Structure 12, 1607–1618.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.011486-0
Loading
/content/journal/jgv/10.1099/vir.0.011486-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error