1887

Abstract

A C57U nucleotide mutation in a predicted RNA stem structure (nt 11–16/56–61) of the 5′ non-coding region (5′NCR) of dengue 2 (DEN-2) 16681 virus is partially attenuating, but unstable during serial passage of certain candidate DEN-2 PDK-53-based vaccine viruses containing this mutation. Here, 11 different mutations (one or more point substitution and/or deletion) between nt 54 and 70 in the 5′NCR of the pD2/IC-30P-A (16681) infectious clone are described. Four mutants were infectious. Three mutants with single point substitutions replicated well in cell culture and exhibited variable neurovirulence in mice. Constructs containing multiple substitutions or any deletions failed to produce infectious viruses. Unexpectedly, a double C57U+G58C mutant replicated as efficiently as D2/IC-30P-A virus, and was more neurovirulent for newborn ICR mice. Thus, despite its predicted additional disruption of the RNA stem structure, the engineered contiguous secondary G58C mutation caused reversion of the partially attenuated phenotype caused by the 5′NCR-C57U mutation.

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2007-06-01
2024-12-02
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References

  1. Ali N., Siddiqui A. 1995; Interaction of polypyrimidine tract-binding protein with the 5′ noncoding region of the hepatitis C virus RNA genome and its functional requirement in internal initiation of translation. J Virol 69:6367–6375
    [Google Scholar]
  2. Alvarez D. E., Lodeiro M. F., Luduena S. J., Pietrasanta L. I., Gamarnik A. V. 2005; Long-range RNA-RNA interactions circularize the dengue virus genome. J Virol 79:6631–6643 [CrossRef]
    [Google Scholar]
  3. Brinton M. A., Dispoto J. H. 1988; Sequence and secondary structure analysis of the 5′-terminal region of flavivirus genome RNA. Virology 162:290–299 [CrossRef]
    [Google Scholar]
  4. Butrapet S., Huang C. Y., Pierro D. J., Bhamarapravati N., Gubler D. J., Kinney R. M. 2000; Attenuation markers of a candidate dengue type 2 vaccine virus, strain 16681 (PDK-53), are defined by mutations in the 5′ noncoding region and nonstructural proteins 1 and 3. J Virol 74:3011–3019 [CrossRef]
    [Google Scholar]
  5. Butrapet S., Kinney R. M., Huang C. Y. 2006; Determining genetic stabilities of chimeric dengue vaccine candidates based on dengue 2 PDK-53 virus by sequencing and quantitative TaqMAMA. J Virol Methods 131:1–9 [CrossRef]
    [Google Scholar]
  6. Cahour A., Pletnev A., Vazielle-Falcoz M., Rosen L., Lai C. J. 1995; Growth-restricted dengue virus mutants containing deletions in the 5′ noncoding region of the RNA genome. Virology 207:68–76 [CrossRef]
    [Google Scholar]
  7. Filomatori C. V., Lodeiro M. F., Alvarez D. E., Samsa M. M., Pietrasanta L., Gamarnik A. V. 2006; A 5′ RNA element promotes dengue virus RNA synthesis on a circular genome. Genes Dev 20:2238–2249 [CrossRef]
    [Google Scholar]
  8. Gubler D. J. 1998; Resurgent vector-borne diseases as a global health problem. Emerg Infect Dis 4:442–450 [CrossRef]
    [Google Scholar]
  9. Guesdon F. M., Greatorex J., Rhee S. R., Fisher R., Hunter E., Lever A. M. 2001; Sequences in the 5′ leader of Mason-Pfizer monkey virus which affect viral particle production and genomic RNA packaging: development of MPMV packaging cell lines. Virology 288:81–88 [CrossRef]
    [Google Scholar]
  10. Huang C. Y., Butrapet S., Pierro D. J., Chang G. J., Hunt A. R., Bhamarapravati N., Gubler D. J., Kinney R. M. 2000; Chimeric dengue type 2 (vaccine strain PDK-53)/dengue type 1 virus as a potential candidate dengue type 1 virus vaccine. J Virol 74:3020–3028 [CrossRef]
    [Google Scholar]
  11. Huang C. Y., Butrapet S., Tsuchiya K. R., Bhamarapravati N., Gubler D. J., Kinney R. M. 2003; Dengue 2 PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development. J Virol 77:11436–11447 [CrossRef]
    [Google Scholar]
  12. Huang C. Y., Silengo S. J., Whiteman M. C., Kinney R. M. 2005; Chimeric dengue 2 PDK-53/West Nile NY99 viruses retain the phenotypic attenuation markers of the candidate PDK-53 vaccine virus and protect mice against lethal challenge with West Nile virus. J Virol 79:7300–7310 [CrossRef]
    [Google Scholar]
  13. Kinney R. M., Butrapet S., Chang G. 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 [CrossRef]
    [Google Scholar]
  14. 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]
  15. Leitmeyer K. C., Vaughn D. W., Watts D. M., Salas R., Villalobos I., de Chacon Ramos C., Rico-Hesse R. 1999; Dengue virus structural differences that correlate with pathogenesis. J Virol 73:4738–4747
    [Google Scholar]
  16. Lindenbach B. D., Rice C. M. 2003; Molecular biology of flaviviruses. Adv Virus Res 59:23–61
    [Google Scholar]
  17. Miller B. R., Mitchell C. J. 1986; Passage of yellow fever virus: its effect on infection and transmission rates in Aedes aegypti . Am J Trop Med Hyg 35:1302–1309
    [Google Scholar]
  18. You S., Falgout B., Markoff L., Padmanabhan R. 2001; In vitro RNA synthesis from exogenous dengue viral RNA templates requires long range interactions between 5′- and 3′-terminal regions that influence RNA structure. J Biol Chem 276:15581–15591 [CrossRef]
    [Google Scholar]
  19. Zuker M. 2003; Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415 [CrossRef]
    [Google Scholar]
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