1887

Abstract

A genomic-length cDNA clone corresponding to the RNA of dengue virus type 2 (DEN-2) New Guinea C strain (NGC) was constructed in a low copy number vector. The cloned cDNA was stably propagated in and designated pDVWS501. RNA transcripts produced from the cDNA using T7 RNA polymerase yielded infectious virus (MON501) upon electroporation into BHK-21 cells. When compared with parental NGC virus, MON501 replicated to similar levels in C6/36 cells and showed similar neurovirulence in suckling mice. In contrast, a second genomic-length cDNA clone (pDVWS310) used as an intermediate in the construction of pDVWS501 produced virus (MON310) that replicated well in C6/36 cells but was not neurovirulent in mice. MON310 contained the prM and E genes of the non-neurovirulent PUO- 218 strain in an NGC background. There were seven amino acid differences between the prM and E proteins of MON310 and MON501. The differences were generally conservative, with the exception of E residue 126, which was Glu in MON310 and Lys in MON501. To examine the role of this residue in mouse neurovirulence, substitutions of Glu → Lys and Lys → Glu were made in MON310and MON501, respectively. The properties of these mutants clearly demonstrated that Lys at E residue 126 is a major determinant of DEN-2 mouse neurovirulence.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-79-3-437
1998-03-01
2022-01-21
Loading full text...

Full text loading...

/deliver/fulltext/jgv/79/3/9519821.html?itemId=/content/journal/jgv/10.1099/0022-1317-79-3-437&mimeType=html&fmt=ahah

References

  1. Anderson C. R., Downs W. G. 1956; Isolation of dengue virus from a human being in Trinidad. Science 124:224–225
    [Google Scholar]
  2. Blok J., Samuel S., Gibbs A. J., Vitarana U. T. 1989; Variation of the nucleotide and encoded amino acid sequences of the envelope gene from eight dengue-2 viruses. Archives of Virology 105:39–53
    [Google Scholar]
  3. Bray M., Lai C.-J. 1991; Construction of intertypic chimeric dengue viruses by substitution of structural protein genes. Proceedings of the National Academy of Sciences, USA 88:10342–10346
    [Google Scholar]
  4. Cecilia D., Gould E. A. 1991; Nucleotide changes responsible for loss of neuroinvasiveness in Japanese encephalitis virus neutralization- resistant mutants. Virology 181:70–77
    [Google Scholar]
  5. Chen W., Kawano H., Men R., Clark D., Lai C.-J. 1995; Construction of intertypic chimeric dengue viruses exhibiting type 3 antigenicity and neurovirulence for mice. Journal of Virology 69:5186–5190
    [Google Scholar]
  6. Deubel V., Kinney R. M., Trent D. W. 1988; Nucleotide sequence and deduced amino acid sequence of the nonstructural proteins of dengue type 2 virus, Jamaica genotype: comparative analysis of the full-length genome. Virology 165:234–244
    [Google Scholar]
  7. Fu J., Tan B.-H., Yap E.-H., Chan Y.-C., Tan Y. H. 1992; Full- length cDNA sequence of dengue type 1 virus (Singapore Strain S275/90). Virology 188:953–958
    [Google Scholar]
  8. Gritsun T. S., Gould E. A. 1995; Infectious transcripts of tick-borne encephalitis virus, generated in days by RT-PCR. Virology 214:611–618
    [Google Scholar]
  9. Gruenberg A., Wright P. J. 1992; Processing of dengue virus type 2 structural proteins containing deletions in hydrophobic domains. Archives of Virology 122:77–94
    [Google Scholar]
  10. Gruenberg A., Woo W. S., Biedrzycka A., Wright P. J. 1988; Partial nucleotide sequence and deduced amino acid sequence of the structural proteins of dengue virus type 2, New Guinea C and PUO-218 strains. Journal of General Virology 69:1391–1398
    [Google Scholar]
  11. Guirakhoo F., Hunt A. R., Lewis J. G., Roehrig J. T. 1993; Selection and partial characterization of dengue 2 virus mutants that induce fusion at elevated pH. Virology 194:219–223
    [Google Scholar]
  12. Hahn C. S., Hahn Y. S., Rice C. M., Lee E., Dalgarno L., Strauss E. G., Strauss J. H. 1987; Conserved elements in the 3ʹ untranslated region of flavivirus RNAs and potential cyclization sequences. Journal of Molecular Biology 198:33–41
    [Google Scholar]
  13. Hahn Y. S., Galler R., Hunkapiller T., Dalrymple J. M., Strauss J. H., Strauss E. G. 1988; Nucleotide sequence of dengue 2 RNA and comparison of the encoded proteins with those of other flaviviruses. Virology 162:167–180
    [Google Scholar]
  14. Hasegawa H., Yoshida M., Shiosaka T., Fujita S., Kobayashi Y. 1992; Mutations in the envelope protein of Japanese encephalitis virus affect entry into cultured cells and virulence in mice. Virology 191:158–165
    [Google Scholar]
  15. Hiramatsu K., Tadano M., Men R., Lai C.-J. 1996; Mutational analysis of a neutralization epitope on the dengue type 2 virus (DEN-2) envelope protein: monoclonal antibody resistant DEN-2/DEN-4 chimeras exhibit reduced mouse neurovirulence. Virology 224:437–445
    [Google Scholar]
  16. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59
    [Google Scholar]
  17. Holzmann H., Heinz F. X., Mandl C. W., Guirakhoo F., Kunz C. 1990; A single amino acid substitution in envelope protein E of tickborne encephalitis virus leads to attenuation in the mouse model. Journal of Virology 64:5156–5159
    [Google Scholar]
  18. Holzmann H., Stiasny K., Ecker M., Kunz C., Heinz F. X. 1997; Characterization of monoclonal antibody-escape mutants of tick-borne encephalitis virus with reduced neuroinvasiveness in mice. Journal of General Virology 78:31–37
    [Google Scholar]
  19. Irie K., Mohan P. M., Sasaguri Y., Putnak R., Padmanabhan R. 1989; Sequence analysis of cloned dengue virus type 2 genome (New Guinea-C strain). Gene 75:197–211
    [Google Scholar]
  20. Jiang W. R., Lowe A., Higgs S., Reid H., Gould E. A. 1993; Single amino acid codon changes detected in louping ill virus antibody-resistant mutants with reduced neurovirulence. Journal of General Virology 74:931–935
    [Google Scholar]
  21. Kapoor M., Zhang L., Mohan P. M., Padmanabhan R. 1995; Synthesis and characterization of an infectious dengue virus type-2 RNA genome (New Guinea C strain). Gene 162:175–180
    [Google Scholar]
  22. Kawano H., Rostapshov V., Rosen L., Lai C.-J. 1993; Genetic determinants of dengue type 4 virus neurovirulence for mice. Journal of Virology 67:6567–6575
    [Google Scholar]
  23. Khromykh A. A., Westaway E. G. 1994; Completion of Kunjin virus RNA sequence and recovery of an infectious RNA transcribed from stably cloned full-length cDNA. Journal of Virology 68:4580–4588
    [Google Scholar]
  24. Kinney R. M., Butrapet S., Chang G.-J. J., Tsuchiya K. R., Roehrig J. T., Bhamarapravati N., Gubler D. J. 1997; Construction of infectious cDNA clones for dengue 2 virus: strain 16681 and its attenuated vaccine derivative, strain PDK-53. Virology 230:300–308
    [Google Scholar]
  25. Lai C.-J., Zhao B., Hori H., Bray M. 1991; Infectious RNA transcribed from stably cloned full-length cDNA of dengue virus type 4. Proceedings of the National Academy of Sciences, USA 88:5139–5143
    [Google Scholar]
  26. Lanciotti R. S., Lewis J. G., Gubler D. J., Trent D. W. 1994; Molecular evolution and epidemiology of dengue-3 viruses. Journal of General Virology 75:65–75
    [Google Scholar]
  27. Lewis J. G., Chang G.-J., Lanciotti R. S., Trent D. W. 1992; Direct sequencing of large flavivirus PCR products for analysis of genome variation and molecular epidemiological investigations. Journal of Viro- logical Methods 38:11–24
    [Google Scholar]
  28. Lewis J. A., Chang G.-J., Lanciotti R. S., Kinney R. M., Mayer L. W., Trent D. W. 1993; Phylogenetic relationships of dengue-2 viruses. Virology 197:216–224
    [Google Scholar]
  29. Lobigs M., Dalgarno L., Schlesinger J. J., Weir R. C. 1987; Location of a neutralization determinant in the E protein of yellow fever virus (17D vaccine strain). Virology 161:474–478
    [Google Scholar]
  30. Lum L. C. S., Lam S. K., Choy Y. S., George R., Harun F. 1996; Dengue encephalitis: a true entity?. American Journal of Tropical Medicine and Hygiene 54:256–259
    [Google Scholar]
  31. McMinn P. C., Marshall I. D., Dalgarno L. 1995; Neurovirulence and neuroinvasiveness of Murray Valley encephalitis virus mutants selected by passage in a monkey kidney cell line. Journal of General Virology 76:865–872
    [Google Scholar]
  32. Mandl C. W., Ecker M., Holzmann H., Kunz C., Heinz F. X. 1997; Infectious cDNA clones of tick-borne encephalitis virus European subtype prototypic strain Neudoerfl and high virulence strain Hypr. Journal of General Virology 78:1049–1057
    [Google Scholar]
  33. Monath T. P. 1994; Dengue: the risk to developed and developing countries. Proceedings of the National Academy of Sciences, USA 91:2395–2400
    [Google Scholar]
  34. Monath T. P., Heinz F. X. 1996; Flaviviruses. In Fields Virology, 3rd edn. 1 pp 961–1034 Fields B. N., Knipe D. M., Howley P. M. Edited by Philadelphia: Lippincott-Raven;
    [Google Scholar]
  35. Murphy F. A., Fauquet C. M., Bishop D. H. L., Ghabrial S. A., Jarvis A. W., Martelli G. P., Mayo M. A., Summers M. D. editors 1995 Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses Vienna & New York: Springer-Verlag;
    [Google Scholar]
  36. Polo S., Ketner G., Levis R., Falgout B. 1997; Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast. Journal of Virology 71:5366–5374
    [Google Scholar]
  37. Rey F. A., Heinz F. X., Mandl C., Kunz C., Harrison S. C. 1995; The envelope glycoprotein from tick-borne encephalitis at 2 Å resolution. Nature 375:291–298
    [Google Scholar]
  38. Rice C. M., Grakoui A., Galler R., Chambers T. J. 1989; Transcription of infectious yellow fever RNA from full-length cDNA templates produced by in vitro ligation. New Biologist 1:285–296
    [Google Scholar]
  39. Sabin A. B., Schlesinger R. W. 1945; Production of immunity to dengue with virus modified by propagation in mice. Science 101:640–642
    [Google Scholar]
  40. Sanchez I. J., Ruiz B. H. 1996; A single nucleotide change in the E protein gene of dengue virus 2 Mexican strain affects neurovirulence in mice. Journal of General Virology 77:2541–2545
    [Google Scholar]
  41. Schlesinger R. W. 1977; Dengue viruses. Virology Monographs 16. Vienna : Springer-Verlag
    [Google Scholar]
  42. Schlesinger R. W., Gordon I., Frankel J. W., Winter J. W., Patterson P. R., Dorrance W. R. 1956; Clinical and serologic response of man to immunization with attenuated dengue and yellow fever viruses. Journal of Immunology 77:352–364
    [Google Scholar]
  43. Shi P.-Y., Brinton M. A., Veal J. M., Zhong Y. Y., Wilson W. D. 1996; Evidence for the existence of a pseudoknot structure at the 3ʹ terminus of the flavivirus genomic RNA. Biochemistry 35:4222–4230
    [Google Scholar]
  44. Shiu S. Y. W., Jiang W. R., Porterfield J. S., Gould E. A. 1992; Envelope protein sequences of dengue virus isolates TH-36 and TH- Sman, and identification of a type-specific genetic marker for dengue and tick-borne flaviviruses. Journal of General Virology 73:207–212
    [Google Scholar]
  45. Stiasny K., Allison S. L., Marchler-Bauer A., Kunz C., Heinz F. X. 1996; Structural requirements for low-pH-induced rearrangements in the envelope glycoprotein of tick-borne encephalitis virus. Journal of Virology 70:8142–8147
    [Google Scholar]
  46. Sumiyoshi H., Hoke C. H., Trent D. W. 1992; Infectious Japanese encephalitis virus RNA can be synthesized from in vitro-ligated cDNA templates. Journal of Virology 66:5425–5431
    [Google Scholar]
  47. Sumiyoshi H., Tignor G. H., Shope R. E. 1995; Characterization of a highly attenuated Japanese encephalitis virus generated from molecularly cloned cDNA. Journal of Infectious Diseases 171:1144–1151
    [Google Scholar]
  48. Trirawatanapong T., Chandran B., Putnak R., Padmanabhan R. 1992; Mapping of a region of dengue virus type-2 glycoprotein required for binding by a neutralizing monoclonal antibody. Gene 116:139–150
    [Google Scholar]
  49. Wang R. F., Kushner S. R. 1991; Construction of versatile low- copy-number vectors for cloning, sequencing and gene expression in Escherichia coli . Gene 100:195–199
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-79-3-437
Loading
/content/journal/jgv/10.1099/0022-1317-79-3-437
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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