1887

Abstract

During virus infection of insect cells, the nucleopolyhedrovirus chitinase is localized primarily within the endoplasmic reticulum (ER), which is consistent with the presence of a carboxy-terminal ER retention motif (KDEL). Release of chitinase into the extracellular medium appears to be concomitant with terminal cell lysis, rather than by active secretion. In this study, we have shown that mutation of the KDEL motif induces a partial redistribution of the chitinase at both early and late times post-infection. Deletion of the KDEL motif or substitution with glycine residues allowed chitinase to move through the secretory pathway, accumulating to detectable levels in the extracellular medium by 24 h post-infection; more than 48 h prior to cell lysis. Deletion of the KDEL motif did not compromise enzyme activity, with the modified enzyme exhibiting characteristic endo- and exo-chitinolytic activity. larvae infected with the modified virus were found to liquefy approximately 24 h earlier than larvae infected with a control virus in which the chitinase KDEL motif had not been deleted.

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2002-03-01
2024-04-18
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References

  1. 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]
  2. Dai X., Hajos J. P., Joosten N. N., van Oers M. M., IJkel W. F. J., Zuidema D., Pang Y., Vlak J. M. 2000; Isolation of a Spodoptera exigua baculovirus recombinant with a 10·6 kbp deletion that retains biological activity. Journal of General Virology 81:2545–2554
    [Google Scholar]
  3. Gomi S., Majima K., Maeda S. 1999; Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. Journal of General Virology 80:1323–1337
    [Google Scholar]
  4. Hawes C. R., Brandizzi F., Andreeva A. V. 1999; Endomembranes and vesicle trafficking. Current Opinion in Plant Biology 2:454–461
    [Google Scholar]
  5. Hawtin R. E., Arnold K., Ayres M. D., Zanotto P. M. D. A., Howard S. C., Gooday G. W., Chappell L. H., Kitts P. A., King L. A., Possee R. D. 1995; Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212:673–685
    [Google Scholar]
  6. 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]
  7. 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]
  8. 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. Journal of General Virology 79:2283–2292
    [Google Scholar]
  9. King L. A., Possee R. D. 1992 The Baculovirus Expression System . A Laboratory Guide London: Chapman and Hall;
    [Google Scholar]
  10. 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]
  11. 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]
  12. McCreath K. J., Gooday G. W. 1992; A rapid and sensitive microassay for determination of chitinolytic activity. Journal of Microbiological Methods 14: pp 229–237
    [Google Scholar]
  13. Ohkawa T., Majima K., Maeda S. 1994; A cysteine protease encoded by the baculovirus Bombyx mor i nuclear polyhedrosis virus. Journal of Virology 68:6619–6625
    [Google Scholar]
  14. Rawlings N. D., Pearl L. H., Buttle D. J. 1992; The baculovirus Autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence. Biology and Chemistry 373:1211–1215
    [Google Scholar]
  15. 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. Journal of General Virology 76:1091–1098
    [Google Scholar]
  16. 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. Journal of Virology 72:10207–10212
    [Google Scholar]
  17. 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. Journal of General Virology 81:1403–1411
    [Google Scholar]
  18. Toyooka K., Okamoto T., Minamikawa T. 2000; Mass transport of Proform of a KDEL-tailed cysteine proteinase (SH-EP) to protein storage vacuoles by endoplasmic reticulum-derived vesicle is involved in protein mobilisation in germinating seeds. Journal of Cell Biology 148:453–464
    [Google Scholar]
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