Deletion of the nucleopolyhedrovirus chitinase KDEL motif and and analysis of the modified virus Free

Abstract

Infection of insect larvae with nucleopolyhedrovirus (AcMNPV) results in the liquefaction of the host, a process involving the action of virus-encoded chitinase and cathepsin gene products. Chitinase is localized in the endoplasmic reticulum (ER) during infection because of the presence of a C-terminal ER retrieval motif (KDEL). In this study, the KDEL coding region was removed from the chitinase gene so that expression of the modified chitinase remained under the control of its own gene promoter, at its native locus. The deletion of KDEL resulted in the redistribution of chitinase within the cell during virus infection. Chitinase lacking the KDEL motif was detectable at the plasma membrane and was also evident in the culture medium of virus-infected cells from as early as 12 h post-infection (p.i.). Secretion of chitinase from the cell continued up to 72 h p.i., until cytolysis. The biological activity of the recombinant virus in larvae was enhanced, with a significant reduction in the lethal dose and lethal time associated with infection. Furthermore, a reduction in feeding damage caused by infected larvae was observed compared to AcMNPV-infected individuals.

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2004-04-01
2024-03-28
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References

  1. Ahrens C. H., Russel R. L., Funk C. J., Evans J. T., Harwood S. H., Rohrmann G. F. 1997; The sequence of the Orygia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. Virology 229:381–399
    [Google Scholar]
  2. Alfonso C. L., Tulman E. R., Lu Z., Balinsky C. A., Moser B. A., Becnel J. J., Rock D. L., Kutish G. F. 2001; Genome sequence of a baculovirus pathogenic for Culex nigripalpus . J Virol 75:11157–11165
    [Google Scholar]
  3. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D. 1994; The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202:586–605
    [Google Scholar]
  4. Bonning B. C., Ward V. K., van Meer M. M. M., Booth T. F., Hammock B. D. 1997; Disruption of lysosomal targeting is associated with insecticidal potency of juvenile hormone esterase. Proc Natl Acad Sci U S A 94:6007–6012
    [Google Scholar]
  5. Bonning B. C., Possee R. D., Hammock B. D. 1999; Insecticidal efficacy of a recombinant baculovirus expressing JHE-KK, a modified juvenile hormone esterase. J Invertebr Pathol 73:234–236
    [Google Scholar]
  6. Chen X., IJkel W. F. J., Tarchini R. 8 other authors 2001; The sequence of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome. J Gen Virol 82:241–257
    [Google Scholar]
  7. Chen X., Zhang W. J., Wong J. 9 other authors 2002; Comparative analysis of the complete genome sequences of Helicoverpa zea and Helicoverpa armigera single-nucleocapsid nucleopolyhedroviruses. J Gen Virol 83:673–684
    [Google Scholar]
  8. Gomi S., Majima K., Maeda S. 1999; Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J Gen Virol 80:1323–1337
    [Google Scholar]
  9. Hammock B. D., Bonning B. C., Possee R. D., Hanzlik T. N., Maeda S. 1990; Expression and effects of the juvenile hormone esterase in a baculovirus vector. Nature 344:458–461
    [Google Scholar]
  10. Harrison R. L., Bonning B. C. 2003; Comparative analysis of the genomes of Rachiplusia ou and Autographa californica multiple nucleopolyhedroviruses. J Gen Virol 84:1827–1842
    [Google Scholar]
  11. Hashimoto Y., Hayakawa T., Ueno Y., Fujita T., Sano Y., Matsumoto T. 2000; Sequence analysis of the Plutella xylostella granulovirus genome. Virology 275:358–372
    [Google Scholar]
  12. Hawes C. R., Brandizzi F., Andreeva A. V. 1999; Endomembranes and vesicle trafficking. Curr Opin Plant Biol 2:454–461
    [Google Scholar]
  13. Hawtin R. E., Arnold K., Ayres M. D. 7 other authors 1995; Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212:673–685
    [Google Scholar]
  14. Hawtin R. E., Zarkowska T., Arnold K., Thomas C. K., Gooday G. W., King L. A., Kuzio J. A., Possee R. D. 1997; Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes. Virology 238:243–253
    [Google Scholar]
  15. Hayakawa T., Ko R., Okano K., Seong S. I., Goto C., Maeda S. 1999; Sequence analysis of the Xestia c-nigrum granulovirus genome. Virology 262:277–297
    [Google Scholar]
  16. Hunter F. R., Crook N. E., Entwistle P. F. 1984; Viruses as pathogens for the control of insects. In Microbial Methods for Environmental Biotechnology pp  323–347 Edited by Grainger J. M., Lynch J. M. New York & London: Academic Press;
    [Google Scholar]
  17. Hyink O., Dellow R. A., Olsen M. J., Caradoc-Davies K. M. B., Drake K., Herniou E. A., Cory J. S., O'Reilly D. R., Ward V. K. 2002; Whole genome analysis of the Epiphyas postvittana nucleopolyhedrovirus. J Gen Virol 83:957–971
    [Google Scholar]
  18. IJkel W. F. J., van Strien E. A., Heldens J. G. M., Broer R., Zuidema D., Goldbach R. W., Vlak J. M. 1999; Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. J Gen Virol 80:3289–3304
    [Google Scholar]
  19. Kang W., Tristem M., Maeda S., Crook N. E., O'Reilly D. R. 1998; Identification and characterization of the Cydia pomonella granulovirus cathepsin and chitinase genes. J Gen Virol 79:2283–2292
    [Google Scholar]
  20. King L. A., Possee R. D. 1992 The Baculovirus Expression System. A Laboratory Guide London: Chapman & Hall;
    [Google Scholar]
  21. Kuzio J., Pearson M. N., Harwood S. H., Funk C. J., Evans J. T., Slavicek J. M., Rohrmann G. F. 1999; Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar . Virology 253:17–34
    [Google Scholar]
  22. Lange M., Jehle J. A. 2003; The genome of the Cryptophlebia leucotreta granulovirus (CrleGV). Virology 317:220–236
    [Google Scholar]
  23. Lewis M. J., Pelham H. R. B. 1992; Ligand-induced redistribution of a human KDEL receptor from the Golgi complex to the endoplasmic reticulum. Cell 68:353–364
    [Google Scholar]
  24. Li Q. J., Donly C., Li L. L., Willis L. G., Theilmann D. A., Erlandson M. 2002; Sequence and organization of the Mamestra configurata nucleopolyhedrovirus genome. Virology 294:106–121
    [Google Scholar]
  25. Luque T., Finch R., Crook N., O'Reilly D. R., Winstanley D. 2001; The complete sequence of the Cydia pomonella granulovirus genome. J Gen Virol 82:2531–2547
    [Google Scholar]
  26. Maeda S., Volrath S. L., Hanzlik T. N., Harper S. A., Majima K., Maddox D. W., Hammock B. D., Fowler E. 1991; Insecticidal effects of an insect-specific neurotoxin expressed by a recombinant baculovirus. Virology 184:777–780
    [Google Scholar]
  27. McCreath K. J., Gooday G. W. 1992; A rapid and sensitive microassay for determination of chitinolytic activity. J Microbiol Methods 14:229–237
    [Google Scholar]
  28. McCutchen B. F., Choudary P. V., Crenshaw R. 7 other authors 1991; Development of a recombinant baculovirus expressing an insect-selective neurotoxin with potential for pest control. Biotechnology 9:848–852
    [Google Scholar]
  29. O'Reilly D. 1997; Auxiliary genes of baculoviruses. In The Baculoviruses pp  267–300 Edited by Miller L. K. New York: Plenum;
    [Google Scholar]
  30. Pang Y., Yu J. X., Wang L. H. 7 other authors 2001; Sequence analysis of the Spodoptera litura multicapsid nucleopolyhedrovirus genome. Virology 287:391–404
    [Google Scholar]
  31. Popham H. J. R., Li Y., Miller L. K. 1997; Genetic improvement of Helicoverpa zea nuclear polyhedrosis virus as a biopesticide. Biol Control 10:83–91
    [Google Scholar]
  32. Possee R. D. 1986; Cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector. Virus Res 5:43–59
    [Google Scholar]
  33. Possee R. D., Howard S. C. 1987; Analysis of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Res 15:10233–10248
    [Google Scholar]
  34. Rawlings N. D., Pearl L. H., Buttle D. J. 1992; The baculovirus Autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence. Biol Chem Hoppe-Seyler 373:1211–1215
    [Google Scholar]
  35. Saville G. P., Thomas C. J., Possee R. D., King L. A. 2002; Partial redistribution of the Autographa californica nucleopolyhedrovirus chitinase in virus-infected cells accompanies mutation of the carboxy-terminal KDEL ER-retrieval motif. J Gen Virol 83:685–694
    [Google Scholar]
  36. Slack J. M., Kuzio J., Faulkner P. 1995; Characterization of v-cath , a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J Gen Virol 76:1091–1098
    [Google Scholar]
  37. Stewart L. M. D., Hirst M., Ferber M. L., Merryweather A. T., Cayley P. J., Possee R. D. 1991; Construction of an improved baculovirus insecticide containing an insect-specific toxin gene. Nature 352:85–88
    [Google Scholar]
  38. Thomas C. J., Brown H. L., Hawes C. R., Lee B. Y., Min M. K., King L. A., Possee R. D. 1998; Localization of a baculovirus-induced chitinase in the insect cell endoplasmic reticulum. J Virol 72:10207–10212
    [Google Scholar]
  39. Thomas C. J., Gooday G. W., King L. A., Possee R. D. 2000; Mutagenesis of the active coding region of the Autographa californica nucleopolyhedrovirus chiA gene. J Gen Virol 81:1403–1411
    [Google Scholar]
  40. Tomalski M. D., Miller L. K. 1991; Insect paralysis by baculovirus-mediated expression of a mite neurotoxin gene. Nature 352:82–85
    [Google Scholar]
  41. Volkman L. E., Zaal K. J. M. 1990; Autographa californica M nuclear polyhedrosis virus: microtubules and replication. Virology 175:292–302
    [Google Scholar]
  42. Winstanley D., Crook N. E. 1993; Replication of Cydia pomonella granulosis virus in cell culture. J Gen Virol 74:1599–1609
    [Google Scholar]
  43. Winstanley D., O'Reilly D. R. 1999; Granuloviruses. In Encyclopaedia of Virology pp  140–146 Edited by Webster R., Granoff A. London: Academic Press;
    [Google Scholar]
  44. Wormleaton S., Kuzio J., Winstanley D. 2003; The complete sequence of the Adoxophyes orana granulovirus genome. Virology 311:350–365
    [Google Scholar]
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