1887

Abstract

The entomopoxviruses (insect poxviruses) of eastern spruce budworm (), two year cycle spruce budworm () and the Indian red army worm () are being studied in our laboratory for their potential as biological insecticides and expression vectors. These viruses characteristically replicate in the cytoplasm of insect cells and produce occlusion bodies that serve to protect the virion from the environment. By analogy to mammalian pox-viruses, they should also contain a viral thymidine kinase (TK) that functions in viral DNA synthesis. The replication of the entomopoxvirus was inhibited by bromodeoxyuridine whereas the baculovirus of was insensitive to this drug. This result was a biochemical indication that entomopoxviruses contained a kinase that phosphorylated this nucleoside analogue and thus viral DNA synthesis was inhibited. TK genes from the three different insect poxviruses were identified, cloned and sequenced. The sequences of the TK genes of the entomopoxviruses were closely related and exhibited 63.2% identity and 9.9% similarity at the protein level. However, there was only 36.7% identity and 13.6% similarity when these enzymes were compared to their mammalian poxvirus counterpart in vaccinia virus. Finally, one entomopoxvirus TK gene was expressed in mutants lacking the enzyme. These bacteria were converted to a phenotype that could incorporate radioactive thymidine into their chromosomal DNA. The results presented in this paper provide impetus for the design of a recombinant entomopoxvirus expression system in which foreign genes could be introduced into the viral TK locus under selective pressure from bromodeoxyuridine.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-73-12-3235
1992-12-01
2022-10-04
Loading full text...

Full text loading...

/deliver/fulltext/jgv/73/12/JV0730123235.html?itemId=/content/journal/jgv/10.1099/0022-1317-73-12-3235&mimeType=html&fmt=ahah

References

  1. Arif M. A. 1984; The entomopoxviruses. Advances in Virus Research 29:195–213
    [Google Scholar]
  2. Arif B. M., Kurstak E. 1991; The entomopoxviruses. In Viruses of Invertebrates pp. 179–195 Edited by Kurstak E. New York: Marcel Dekker Publishers;
    [Google Scholar]
  3. Banville M., Dumas F., Trifiro S., Arif B., Richardson C. 1992; The predicted amino acid sequence of the spheroidin protein from Amsacta moorei entomopoxvirus: lack of homology between major occlusion body proteins of different poxviruses. Journal of General Virology 73:559–566
    [Google Scholar]
  4. Binns M. M., Tomley F. M., Campbell J., Boursnell M. E. G. 1988; Comparison of a conserved region in fowlpox virus and vaccinia virus genomes and the translocation of the fowlpox virus thymidine kinase gene. Journal of General Virology 69:1275–1283
    [Google Scholar]
  5. Black M. E., Hruby D. E. 1990; Identification of the ATP-binding domain of vaccinia virus thymidine kinase. Journal of Biological Chemistry 265:17584–17592
    [Google Scholar]
  6. Boyle D. B., Coupar B. E., Gibbs A. J., Seigman L. J., Both G. W. 1987; Fowlpox virus thymidine kinase: nucleotide sequence and relationships to other thymidine kinases. Virology 156:355–365
    [Google Scholar]
  7. Bradshaw H. D., Denninger P. L. 1984; Human thymidine kinase gene: molecular cloning and nucleotide sequence of a cDNA expressible in mammalian cells. Molecular and Cellular Biology 4:2316–2320
    [Google Scholar]
  8. Davison A. J., Moss B. 1989; Structure of vaccinia virus early promoters. Journal of Molecular Biology 210:749–769
    [Google Scholar]
  9. Esposito J. J., Knight J. C. 1984; Nucleotide sequence of the thymidine kinase gene region of monkeypox and variola viruses. Virology 135:561–567
    [Google Scholar]
  10. Feller J. A., Massung R. F., Turner P. C., Gibbs E. P. J., Bockamp E. O., Beloso A., Talavera A., Vinuela E., Moyer R. W. 1991; Isolation and molecular characterization of the swinepox virus thymidine kinase gene. Virology 183:578–585
    [Google Scholar]
  11. Gershon P. D., Black D. N. 1989; The nucleotide sequence around the capripoxvirus thymidine kinase gene reveals a gene shared specifically with leporipoxvirus. Journal of General Virology 70:525–533
    [Google Scholar]
  12. Goodwin R. H., Adams J. R., Shapiro M. 1990; Replication of the entomopoxvirus from Amsacta moorei in serum-free cultures of a gypsy moth cell line. Journal of Invertebrate Pathology 56:190–205
    [Google Scholar]
  13. Granados R. R., Roberts D. W. 1970; Electron microscopy of a poxlike virus infecting an invertebrate host. Virology 40:230–243
    [Google Scholar]
  14. Gruidl M. E., Hall R. L., Moyer R. W. 1992; Mapping and molecular characterization of a functional thymidine kinase from Amsacta moorei entomopoxvirus. Virology 186:507–516
    [Google Scholar]
  15. Hall R. L., Moyer R. W. 1991; Identification, cloning, and sequencing of a fragment of Amsacta moorei entomopoxvirus DNA containing the spheroidin gene and three vaccinia virus-related open reading frames. Journal of Virology 65:6516–6527
    [Google Scholar]
  16. Hiraga S., Igarashi K., Yura T. 1967; A deoxythymidine kinase-deficient mutant of E.coli . Biochimica et biophysica acta 145:41–51
    [Google Scholar]
  17. Kit S. 1985; Thymidine kinase. Microbiological Sciences 2:369–375
    [Google Scholar]
  18. Kit S., Dubbs D. R., Frearson P. M. 1966; HeLa cells resistant to bromodeoxyuridine and deficient in thymidine kinase activity. International Journal of Cancer 1:19–30
    [Google Scholar]
  19. Lin P. F., Lieberman H. B., Yeh D. B., Xu T., Zhao S. Y., Ruddle F. H. 1985; Molecular cloning and structural analysis of murine thymidine kinase genomic and cDNA sequences. Molecular and Cellular Biology 5:3149–3156
    [Google Scholar]
  20. Liu K. Y. C., Manning J. S. 1986; Identification of the thymidine kinase gene of infectious bovine rhinotracheitis virus and its function in Escherichia coli hosts. Gene 44:279–285
    [Google Scholar]
  21. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  22. Mackett M., Smith G. L., Moss B. 1982; Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proceedings of the National Academy of Sciences, U.S.A. 79:7415–7419
    [Google Scholar]
  23. Martin Hernandez A. M., Tabares E. 1991; Expression and characterization of the thymidine kinase gene of African swine fever virus. Journal of Virology 65:1046–1052
    [Google Scholar]
  24. Miller L. K. 1988; Baculoviruses as gene expression vectors. Annual Reviews of Microbiology 42:177–199
    [Google Scholar]
  25. Panicali D., Paoletti E. 1982; Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccinia virus. Proceedings of the National Academy of Sciences, U.S.A. 79:4927–4931
    [Google Scholar]
  26. Pearson A., Richardson C., Yuen L. 1991; The 5′ non-coding region sequence of the Choristoneura biennis entomopoxvirus spheroidin gene functions as an efficient late promoter in mammalian vaccinia expression system. Virology 180:561–566
    [Google Scholar]
  27. Roizman B., Jenkins F. J. 1985; Genetic engineering of novel genomes of large DNA viruses. Science 229:1208–1214
    [Google Scholar]
  28. Schnitzlein W. M., Tripathy D. N. 1991; Identification and nucleotide sequence of the thymidine kinase gene of swinepox virus. Virology 181:727–732
    [Google Scholar]
  29. Upton C., McFadden G. 1986; Identification and nucleotide sequence of the thymidine kinase gene of Shope fibroma virus. Journal of Virology 60:920–927
    [Google Scholar]
  30. Vialard J., Lalumiere M., Vernet T., Briedis D., Alkhatib G., Henning D., Levin D., Richardson C. 1990; Synthesis of the membrane fusion and hemagglutinin proteins of measles virus, using a novel baculovirus vector containing the β-galactosidase gene. Journal of Virology 64:765–776
    [Google Scholar]
  31. Weir J. P., Moss B. 1983; Nucleotide sequence of the vaccinia virus thymidine kinase gene and the nature of spontaneous frameshift mutations. Journal of Virology 46:530–537
    [Google Scholar]
  32. Yuen L., Moss B. 1987; Oligonucleotide sequence signaling transcriptional termination of vaccinia virus early genes. Proceedings of the National Academy of Sciences, U.S.A. 84:6417–6421
    [Google Scholar]
  33. Yuen L., Dionne J., Arif B., Richardson C. 1990; Identification and sequencing of the spheroidin gene of Choristoneura biennis entomopoxvirus. Virology 175:427–433
    [Google Scholar]
  34. Yuen L., Noiseux M., Gomes M. 1991; DNA sequence of the nucleoside triphosphate phosphohydrolase I (NPH I) of the Choristoneura biennis entomopoxvirus. Virology 182402–406
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-73-12-3235
Loading
/content/journal/jgv/10.1099/0022-1317-73-12-3235
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