1887

Abstract

Post-mortem host degradation by infection of nucleopolyhedrovirus (BmNPV) requires the synergistic activation of two virus-encoded genes, () and (). Previous studies have suggested that V-CHIA is essential for the proper folding of the nascent V-CATH polypeptide in the endoplasmic reticulum, and that the putative V-CHIA–V-CATH interaction might be mediated by -linked glycans of V-CATH. Sequence analysis shows that BmNPV V-CATH includes three consensus -linked glycosylation sites (asparagine 38, 65 and 158). To clarify the role of -linked glycans of V-CATH in its biological activity, we generated three recombinant BmNPVs expressing mutant V-CATHs, and found that the two residues, asparagine 38 and 65, which are localized in the pro-region of V-CATH, are the glycosylation sites of BmNPV V-CATH. Western blot analysis also showed that removal of -linked glycans from BmNPV V-CATH resulted in production of the insoluble forms of V-CATH and V-CHIA. These results demonstrate that -linked glycans located in the pro-region of BmNPV V-CATH are essential for the proper folding of V-CATH and V-CHIA.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.005835-0
2009-01-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/1/170.html?itemId=/content/journal/jgv/10.1099/vir.0.005835-0&mimeType=html&fmt=ahah

References

  1. Brömme, D. & Okamoto, K.(1995). The baculovirus cysteine protease has a cathepsin B-like S2-subsite specificity. Biol Chem Hoppe Seyler 376, 611–615.[CrossRef] [Google Scholar]
  2. Daimon, T., Katsuma, S. & Shimada, T.(2007). Mutational analysis of active site residues of chitinase from Bombyx mori nucleopolyhedrovirus. Virus Res 124, 168–175.[CrossRef] [Google Scholar]
  3. Federici, B. A.(1997). Baculovirus Pathogenesis. In The Baculoviruses, pp. 33–59. Edited by L. K. Miller. New York: Plenum.
  4. Hawtin, R. E., Arnold, K., Ayres, M. D., Zanotto, P. M., 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.[CrossRef] [Google Scholar]
  5. Hawtin, R. E., Zarkowska, T., Arnold, K., Thomas, C. J., 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.[CrossRef] [Google Scholar]
  6. Hom, L. G. & Volkman, L. E.(2000).Autographa californica M nucleopolyhedrovirus chiA is required for processing of V-CATH. Virology 277, 178–183.[CrossRef] [Google Scholar]
  7. Hong, H. K., Woo, S. D., Choi, J. Y., Lee, H. K., Kim, M. H., Je, Y. H. & Kang, S. K.(2000). Characterization of four isolates of Bombyx mori nucleopolyhedrovirus. Arch Virol 145, 2351–2361.[CrossRef] [Google Scholar]
  8. Jarvis, D. L., Wills, L., Burow, G. & Bohlmeyer, D. A.(1998). Mutational analysis of the N-linked glycans on Autographa californica nucleopolyhedrovirus gp64. J Virol 72, 9459–9469. [Google Scholar]
  9. Katsuma, S., Shimada, T. & Kobayashi, M.(2004a). Characterization of the baculovirus Bombyx mori nucleopolyhedrovirus gene homologous to the mammalian FGF gene family. Virus Genes 29, 211–217.[CrossRef] [Google Scholar]
  10. Katsuma, S., Tanaka, S., Shimada, T. & Kobayashi, M.(2004b). Reduced cysteine protease activity of the hemolymph of Bombyx mori larvae infected with fp25K-inactivated Bombyx mori nucleopolyhedrovirus results in the reduced postmortem host degradation. Arch Virol 149, 1773–1782. [Google Scholar]
  11. Katsuma, S., Daimon, T., Mita, K. & Shimada, T.(2006a). Lepidopteran ortholog of Drosophila Breathless is a receptor for the baculovirus fibroblast growth factor. J Virol 80, 5474–5481.[CrossRef] [Google Scholar]
  12. Katsuma, S., Daimon, T., Horie, S., Kobayashi, M. & Shimada, T.(2006b). N-linked glycans of Bombyx mori nucleopolyhedrovirus fibroblast growth factor are crucial for its secretion. Biochem Biophys Res Commun 350, 1069–1075.[CrossRef] [Google Scholar]
  13. Long, G., Pan, X. & Vlak, J. M.(2007). Absence of N-linked glycans from the F2 subunit of the major baculovirus envelope fusion protein F enhances fusogenicity. J Gen Virol 88, 441–449.[CrossRef] [Google Scholar]
  14. Ohkawa, T., Majima, K. & Maeda, S.(1994). A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. J Virol 68, 6619–6625. [Google Scholar]
  15. O'Reilly, D. R.(1997). Auxiliary genes of baculoviruses. In The Baculoviruses, pp. 267–300. Edited by L. K. Miller. New York: Plenum.
  16. 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.[CrossRef] [Google Scholar]
  17. Wang, F., Zhang, C. X., Kumar, V. S. & Wu, X. F.(2005). Influences of chitinase gene deletion from BmNPV on the cell lysis and host liquefaction. Arch Virol 150, 981–990.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.005835-0
Loading
/content/journal/jgv/10.1099/vir.0.005835-0
Loading

Data & Media loading...

Supplements

List of primers [PDF](16 KB)

PDF

[Single PDF](623 KB)

PDF
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