1887

Abstract

The genome of infectious pancreatic necrosis virus (IPNV) is composed of two segments of dsRNA. The larger segment contains a small ORF partly overlapping the 5′ end of the polyprotein reading frame. Yet very little is known about this possible new gene, which presumably codes for a 17 kDa polypeptide (VP5). The region of the viral genome which encompasses the small ORF was reverse-transcribed and amplified by PCR before cloning and sequencing. Analysis of the sequences obtained from five different virus strains revealed that the small ORF is not found on one of them, and that it is truncated on two others. Moreover, the deduced amino acid sequences did not appear to be well conserved. Despite the large variations between IPNV strains at the genomic level, all predicted VP5 are arginine-rich basic polypeptides. To verify whether the small ORF is translated into protein in fish cells, the 17 kDa polypeptide of the VR-299 strain was expressed as a fusion protein in a prokaryotic expression vector and used to produce a specific antiserum. This antiserum reacted with concentrated virus in an immunodot assay, indicating that VP5 is synthesized in infected cells, but probably only in small quantities. When tested with 12 other IPNV strains, results were less conclusive than those obtained with strain VR-299. Nevertheless, three of the 12 viruses gave a clearly negative signal in the immunodot assay, suggesting that possibly more than one viral strain lacks the small ORF.

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1995-08-01
2021-10-22
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References

  1. Berthiaume L., Tarrab E., Heppell J., Arella M., Dobos P., Duncan R., Lecomte J. 1992; Antigenic and genomic differences of two Jasper strains of infectious pancreatic necrosis virus. Intervirology 34:197–201
    [Google Scholar]
  2. Chung H. K. Lee H. S., Lee H. H., Ha Y. C., Lee D. S., Kim Y. S. 1993; Nucleotide sequence analysis of genome segment A of infectious pancreatic necrosis virus DRT strain. GenBank accession number D26526
    [Google Scholar]
  3. Cohen J. 1975; Ribonucleic acid polymerase activity in purified infectious pancreatic necrosis virus of trout. Biochemical and Biophysical Research Communications 62:689–695
    [Google Scholar]
  4. Dobos P., Hill B. J., Hallett R., Kells D. T. C., Becht H., Teninges D. 1979; Biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded RNA genomes. Journal of Virology 32:593–605
    [Google Scholar]
  5. Dobos P., Nagy E., Duncan R. 1991; Birnaviridae. In Viruses of Invertebrates pp 301–314 Edited by Kurstak E. New York: Marcel Dekker;
    [Google Scholar]
  6. Duncan R., Dobos P. 1986; The nucleotide sequence of infectious pancreatic necrosis virus (IPNV) dsRNA segment A reveals one large ORF encoding a precursor polyprotein. Nucleic Acids Research 14:5934
    [Google Scholar]
  7. Duncan R., Nagy E., Krell P. J., Dobos P. 1987; Synthesis of the infectious pancreatic necrosis virus polyprotein, detection of a virus-encoded protease, and fine structure mapping of genome segment A coding regions. Journal of Virology 61:3655–3664
    [Google Scholar]
  8. Duncan R., Mason C. L., Nagy E., Leong J., Dobos P. 1991; Sequence analysis of infectious pancreatic necrosis virus genome segment B and its encoded VP1 protein: a putative RNA-dependant RNA polymerase lacking the Gly–Asp–Asp motif. Virology 181:541–552
    [Google Scholar]
  9. Ernst H., Shatkin A. J. 1985; Reovirus hemagglutinin mRNA codes for two polypeptides in overlapping reading frames. Proceedings of the National Academy of Sciences, USA 82:48–52
    [Google Scholar]
  10. Francki R. I. B., Fauquet C. M., Knudson D. L., Brown F. 1991; Classification and Nomenclature of Viruses. Fifth report of the International Committee on Taxonomy of Viruses. Archives of Virology Supplementum 2.
    [Google Scholar]
  11. Håvarstein L. S., Kalland K. H., Christie K. E., Endresen C. 1990; Sequence of the large double-stranded RNA segment of the N1 strain of infectious pancreatic necrosis virus: a comparison with other Birnaviridae . Journal of General Virology 71:299–308
    [Google Scholar]
  12. Heppell J., Berthiaume L., Tarrab E., Lecomte J., Arella M. 1992; Evidence of genomic variations between infectious pancreatic necrosis virus strains determined by restriction fragment profiles. Journal of General Virology 73:2863–2870
    [Google Scholar]
  13. Heppell J., Berthiaume L., Corbin F., Tarrab E., Lecomte J., Arella M. 1993; Comparison of amino acid sequences deduced from a cDNA fragment obtained from infectious pancreatic necrosis virus (IPNV) strains of different serotypes. Virology 195:840–844
    [Google Scholar]
  14. Huang M. T. F., Manning D. S., Warner M., Stephens E. B., Leong J. C. 1986; A physical map of the viral genome for infectious pancreatic necrosis virus Sp: analysis of cell-free translation products derived from viral cDNA clones. Journal of Virology 60:1002–1011
    [Google Scholar]
  15. Kibenge F. S. B., Jackwood D. J., Mercado C. C. 1990; Nucleotide sequence analysis of genome segment A of infectious bursal disease virus. Journal of General Virology 71:569–577
    [Google Scholar]
  16. Kozak M. 1987; An analysis of 5-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Research 15:8125–8132
    [Google Scholar]
  17. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157:105–132
    [Google Scholar]
  18. MacDonald R. D., Dobos P. 1981; Identification of the protein encoded by each genome segment of infectious pancreatic necrosis virus. Virology 114:414–422
    [Google Scholar]
  19. Magyar G., Dobos P. 1994a; Expression of infectious pancreatic necrosis virus polyprotein and VP1 in insect cells and the detection of the polyprotein in purified virus. Virology 198:437–445
    [Google Scholar]
  20. Magyar G., Dobos P. 1994b; Evidence for the detection of the infectious pancreatic necrosis virus polyprotein and the 17-kDa polypeptide in infected cells and of the NS protease in purified virus. Virology 204:580–589
    [Google Scholar]
  21. Manning D. S., Leong J. C. 1990; Expression in Escherichia coli of the large genomic segment of infectious pancreatic necrosis virus. Virology 179:16–25
    [Google Scholar]
  22. Manning D. S., Mason C. L., Leong J. C. 1990; Cell-free translational analysis of the processing of infectious pancreatic necrosis virus polyprotein. Virology 179:9–15
    [Google Scholar]
  23. Nagy E., Duncan R., Krell P., Dobos P. 1987; Mapping of the large RNA genome segment of infectious pancreatic necrosis virus by hybrid arrested translation. Virology 158:211–217
    [Google Scholar]
  24. Sanger F., Nickxen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74:5463–5467
    [Google Scholar]
  25. Spiropoulou C. F., Nichol S. T. 1993; A small highly basic protein is encoded in overlapping frame within the P gene of vesicular stomatitis virus. Journal of Virology 67:3103–3110
    [Google Scholar]
  26. Tarrab E., Berthiaume L., Heppell J., Arella M., Lecomte J. 1993; Antigenic characterization of serogroup ‘A’ of infectious pancreatic necrosis virus with three panels of monoclonal antibodies. Journal of General Virology 74:2025–2030
    [Google Scholar]
  27. Wolf K. 1988 Fish Viruses and Fish Viral Diseases Ithaca: Cornell University Press.;
    [Google Scholar]
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