1887

Abstract

The multiple nucleopolyhedrovirus (MNPV) protein p74 is associated with the occlusion-derived virus (ODV) envelope. p74 is essential for oral infectivity of ODV and has been proposed to play a role in midgut attachment and/or fusion. In this study, p74 protein was expressed in-frame with green fluorescent protein (GFP) to create a p74–GFP chimera. The C-terminal GFP portion of the chimera facilitated visualization of the trafficking of p74 in baculovirus-infected (-9) cells. p74–GFP chimeric proteins localized in the intranuclear ring zone of the nucleus and were found to co-precipitate with the microvesicle fraction of cell lysates. A series of truncations of p74 was expressed as p74–GFP chimeras in recombinant baculoviruses. When C-terminal region S580–F645 was deleted from p74, p74–GFP chimera localization became non-specific and chimeras became soluble. p74 region S580–F645 directed GFP to the intranuclear ring zone in a similar pattern to full-length p74. The hydrophobic C terminus of p74 plays a role in protein localization and possibly in transmembrane anchoring and insertion.

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2001-09-01
2024-11-03
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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 protein of the occlusion-derived virus envelope: ODV-E56. Virology 217:97–110
    [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
    [Google Scholar]
  3. Chang M. J., Kuzio J., Blissard G. W. 1999; Modulation of translational efficiency by contextual nucleotides flanking a baculovirus initiator AUG codon. Virology 5:369–383
    [Google Scholar]
  4. Dignam J. D. 1990; Preparation of extracts from higher eukaryotes. In Guide to Protein Purification pp 194–203 Edited by Deutscher M. P. New York & London: Academic Press;
    [Google Scholar]
  5. Faulkner P., Carstens E. B., Eaton B. T., Cochran M. A. 1982; Molecular cloning and physical mapping of restriction endonuclease fragments of Autographa californica nuclear polyhedrosis virus DNA. Journal of Virology 41:940–946
    [Google Scholar]
  6. 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. Journal of General Virology 78:3091–3100
    [Google Scholar]
  7. Fonseca F. G., Wolffe E. J., Weisberg A., Moss B. 2000; Characterization of the vaccinia virus H3L envelope protein: topology and posttranslational membrane insertion via the C-terminal hydrophobic tail. Journal of Virology 74:7508–7517
    [Google Scholar]
  8. Granados R. R., Lawler K. A. 1981; In vivo pathway of Autographa californica baculovirus invasion and infection. Virology 108:297–308
    [Google Scholar]
  9. 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
    [Google Scholar]
  10. 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]
  11. Higgins D. G., Sharp P. M. 1989; Fast and sensitive multiple sequence alignments on a microcomputer. Computer Applications in the Biosciences 2:151–153
    [Google Scholar]
  12. Hink W. F. 1970; Established insect cell line from the cabbage looper, Trichoplusia ni. Nature 226:466–467
    [Google Scholar]
  13. Hink W. F., Vail P. V. 1973; A plaque assay for titration of alfalfa looper nuclear polyhedrosis virus in the cabbage looper (TN-368) cell line. Journal of Invertebrate Pathology 22:168–174
    [Google Scholar]
  14. Hohmann A. W., Faulkner P. 1983; Monoclonal antibodies to baculovirus structural proteins: determination of specificities by western blot analysis. Virology 125:432–444
    [Google Scholar]
  15. Hong T., Braunagel S. C., Summers M. D. 1994; Transcription, translation, and cellular localization of PDV-E66: a structural protein of the PDV envelope of Autographa californica nuclear polyhedrosis virus. Virology 204:210–222
    [Google Scholar]
  16. Hong T., Summers M. D., Braunagel S. C. 1997; N-terminal sequences from Autographa californica nuclear polyhedrosis virus envelope proteins ODV-E66 and ODV-E25 are sufficient to direct reporter proteins to the nuclear envelope, intranuclear microvesicles, and the envelope of occlusion-derived virus. Proceedings of the National Academy of Sciences, USA 94:4050–4055
    [Google Scholar]
  17. Horton H. M., Burand J. P. 1993; Saturable attachment sites for polyhedron-derived baculovirus on insect cells and evidence for entry via direct membrane fusion. Journal of Virology 67:1860–1868
    [Google Scholar]
  18. Kutay U., Hartmann E., Rapoport T. A. 1993; A class of membrane proteins with a C-terminal anchor. Trends in Cell Biology 3:72–75
    [Google Scholar]
  19. Kuzio J., Jaques R., Faulkner P. 1989; Identification of p74 a gene essential for virulence of baculovirus occlusion bodies. Virology 173:759–763
    [Google Scholar]
  20. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157:105–132
    [Google Scholar]
  21. Moser B. A., Becnel J. J., White S. E., Afonso C., Kutish G., Shanker S., Almir E. 2001; Morphological and molecular evidence that Culex nigripalpus baculovirus is an unusual member of the family Baculoviridae. Journal of General Virology 82:283–397
    [Google Scholar]
  22. Oomens A. G. P., Blissard G. W. 1999; Requirement for GP64 to drive efficient budding of Autographa californica multicapsid nucleopolyhedrovirus. Virology 254:297–314
    [Google Scholar]
  23. Russell R. L. Q., Rohrmann G. F. 1993; A 25-kda protein is associated with the envelopes of occluded baculovirus virions. Virology 195:532–540
    [Google Scholar]
  24. Slack J. M., Blissard G. W. 1997; Identification of two independent transcriptional activation domains in the Autographa californica multicapsid nuclear polyhedrosis virus IE1 protein. Journal of Virology 71:9579–9587
    [Google Scholar]
  25. 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]
  26. Sonnhammer E. L., von Heijne G., Krogh A. 1998; A hidden Markov model for predicting transmembrane helices in protein sequences. Intelligent Systems for Molecular Biology 6:175–182
    [Google Scholar]
  27. Stoltz D. B., Pavan C., Dacunha A. B. 1973; Nuclear polyhedrosis virus, a possible example of de novo intranuclear membrane morphogenesis. Journal of General Virology 19:145–150
    [Google Scholar]
  28. Summers M. D., Volkman L. E. 1976; Comparison of biophysical and morphological properties of occluded and extracellular non-occluded baculovirus from in vivo and in vitro host systems. Journal of Virology 17:962–972
    [Google Scholar]
  29. Talcott B., Moore M. S. 1999; Getting across the nuclear pore complex. Trends in Cell Biology 8:312–318
    [Google Scholar]
  30. Theilmann D. A., Chantler J. K., Stewart S., Flipsen H. T. M., Vlak J. M., Crook N. E. 1996; Characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions. Virology 218:148–158
    [Google Scholar]
  31. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. 1977; The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae. In Vitro 13:213–217
    [Google Scholar]
  32. Whitford M., Stewart S., Kuzio J., Faulkner P. 1989; Identification and sequence analysis of a gene encoding gp67 , an abundant envelope glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus. Journal of Virology 63:1393–1399
    [Google Scholar]
  33. Yoshifumi H., 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]
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