1887

Abstract

Baculovirus occlusion-derived virions (ODVs) are released from occlusion bodies by the alkaline environment of the insect midgut. The ODV envelope protein P74 is required for oral infectivity. A soluble form of the P74 protein, P74, was engineered as part of a chimeric protein with jellyfish green fluorescent protein (GFP). P74–GFP was overproduced by the baculovirus expression system and purified away from the wild-type P74. Brush border membrane vesicles (BBMVs) were prepared from the midguts of third-instar larvae. When P74–GFP was incubated under alkaline conditions with BBMVs, a P74–GFP product with a smaller molecular mass was produced. Immunoblots indicated that the smaller product was generated by N-terminal cleavage of P74. This cleavage was prevented by soybean trypsin inhibitor. Analysis of the peptide sequences of P74 homologues identified a conserved trypsin cleavage site that could generate the observed P74–GFP BBMV-specific cleavage product.

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2005-06-01
2019-12-15
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References

  1. Braunagel, S. C., Elton, D. M., Ma, H. & Summers, M. D. ( 1996a; ). Identification and analysis of an Autographa californica nuclear polyhedrosis virus structural proteins of the occlusion-derived virus envelope: ODV-E56. Virology 217, 97–110.[CrossRef]
    [Google Scholar]
  2. Braunagel, S. C., He, H., Ramamurthy, P. & Summers, M. D. ( 1996b; ). Transcription, translation, and cellular localization of three Autographa californica nuclear polyhedrosis virus structural proteins: ODV-E18, ODV-E35, and ODV-EC27. Virology 222, 100–114.[CrossRef]
    [Google Scholar]
  3. Cheng, C.-H., Liu, S.-M., Chow, T.-Y., Hsiao, Y.-Y., Wang, D.-P., Huang, J.-J. & Cheng, H.-H. ( 2002; ). Analysis of the complete genome sequence of the Hz-1 virus suggests that it is related to the members of the Baculoviridae. J Virol 76, 9024–9034.[CrossRef]
    [Google Scholar]
  4. Faulkner, P., Kuzio, J., Williams, G. V. & Wilson, J. A. ( 1997; ). Analysis of p74, a PDV envelope protein of Autographa californica nucleopolyhedrovirus required for occlusion body infectivity in vivo. J Gen Virol 78, 3091–3100.
    [Google Scholar]
  5. Haas-Stapleton, E. J., Washburn, J. O. & Volkman, L. E. ( 2004; ). P74 mediates specific binding of Autographa californica M nucleopolyhedrovirus occlusion-derived virus to primary cellular targets in the midgut epithelia of Heliothis virescens larvae. J Virol 78, 6786–6791.[CrossRef]
    [Google Scholar]
  6. Hefferon, K. L., Oomens, A. G., Monsma, S. A., Finnerty, C. M. & Blissard, G. W. ( 1999; ). Host cell receptor binding by baculovirus GP64 and kinetics of virion entry. Virology 258, 455–468.[CrossRef]
    [Google Scholar]
  7. Hink, W. F. ( 1970; ). Established insect cell line from the cabbage looper, Trichoplusia ni. Nature 226, 466–467.
    [Google Scholar]
  8. Horton, H. M. & Burand, J. P. ( 1993; ). Saturatable attachment sites for polyhedron-derived baculoviruses on insect cells and evidence for entry via direct membrane fusion. J Virol 67, 1860–1868.
    [Google Scholar]
  9. Johnston, K. A., Lee, M. J., Gatehouse, J. A. & Anstee, J. H. ( 1991; ). The partial purification and characterization of serine protease activity in Helicoverpa armigera. Insect Biochem 21, 389–397.[CrossRef]
    [Google Scholar]
  10. Kikhno, I., Gutierrez, S., Croizier, L., Croizier, G. & Ferber, M. L. ( 2002; ). Characterization of pif, a gene required for the per os infectivity of Spodoptera littoralis nucleopolyhedrovirus. J Gen Virol 83, 3013–3022.
    [Google Scholar]
  11. Kitts, P. A. & Possee, R. D. ( 1993; ). A method for producing recombinant baculovirus expression vectors at high frequency. Biotechniques 14, 810–817.
    [Google Scholar]
  12. Kuzio, J., Jaques, R. & Faulkner, P. ( 1989; ). Identification of p74, a gene essential for virulence of baculovirus occlusion bodies. Virology 173, 759–763.[CrossRef]
    [Google Scholar]
  13. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  14. Lung, O., Westenberg, M., Vlak, J. M., Zuidema, D. & Blissard, G. W. ( 2002; ). Pseudotyping Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV): F proteins from group II NPVs are functionally analogous to AcMNPV GP64. J Gen Virol 76, 5729–5736.[CrossRef]
    [Google Scholar]
  15. Monsma, S. A., Oomens, A. G. & Blissard, G. W. ( 1996; ). The GP64 envelope fusion protein is an essential baculovirus protein required for cell-to-cell transmission of infection. J Virol 70, 4607–4616.
    [Google Scholar]
  16. Pijlman, G. P., Pruijssers, A. J. & Vlak, J. M. ( 2003; ). Identification of pif-2, a third conserved baculovirus gene required for per os infection of insects. J Gen Virol 84, 2041–2049.[CrossRef]
    [Google Scholar]
  17. Rukmini, V., Reddy, C. Y. & Venkateswerlu, G. ( 2000; ). Bacillus thuringiensis crystal delta-endotoxin: role of proteases in the conversion of protoxin to toxin. Biochimie 82, 109–116.[CrossRef]
    [Google Scholar]
  18. Slack, J. M., Dougherty, E. M. & Lawrence, S. D. ( 2001; ). A study of the Autographa californica multiple nucleopolyhedrovirus ODV envelope protein p74 using a GFP tag. J Gen Virol 82, 2279–2287.
    [Google Scholar]
  19. Terra, W. R. & Ferreira, C. ( 1994; ). Insect digestive enzymes: properties, compartmentalization and function. Comp Biochem Physiol 109B, 1–62.
    [Google Scholar]
  20. Theilmann, D. A., Chantler, J. K., Stewart, S., Flipsen, T. M., Vlak, J. M. & Crooks, N. E. ( 1996; ). Characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions. Virology 218, 148–158.[CrossRef]
    [Google Scholar]
  21. Westenberg, M., Veenman, F., Roode, E. C., Goldbach, R. W., Vlak, J. M. & Zuidema, D. ( 2004; ). Functional analysis of the putative fusion domain of the baculovirus envelope fusion protein F. J Virol 78, 6946–6954.[CrossRef]
    [Google Scholar]
  22. Wolfersberger, M. G., Luethy, P., Maurer, A., Parenti, P., Sacchi, F. V., Giordana, B. & Hanozet, G. M. ( 1987; ). Preparation and partial characterization of amino acid transporting brush border membrane vesicles from the larval midgut of the cabbage butterfly (Pieris brassicae). Comp Biochem Physiol 86A, 301–308.
    [Google Scholar]
  23. Yao, L., Zhou, W., Xu, H., Zheng, Y. & Qi, Y. ( 2004; ). The Heliothis armigera single nucleocapsid nucleopolyhedrosis envelope protein P74 is required for infection of the host midgut. Virus Res 104, 111–121.[CrossRef]
    [Google Scholar]
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