1887

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Volume 74, Issue 4

Functional domains of the p10 protein of nuclear polyhedrosis virus Free

Abstract

Distinct functional domains in the nuclear polyhedrosis virus p10 protein were identified by analysis of p10 mutants. When up to 15 amino acids from the carboxy terminus were deleted, truncated p10 proteins were found in both the nucleus and the cytoplasm of infected cells, but formed no fibrillar structures. This suggested that the positively charged carboxy terminus is not required for nuclear or cytoplasmic localization of p10 protein, but is involved in protein–protein interactions leading to assembly of the p10 protein into fibrillar structures. Absence of the p10 protein prevented the release of polyhedra from infected cells, caused by impaired nuclear disintegration. This function of the p10 protein appears to be located between amino acid residues 52 and 79. The aminoterminal half of the p10 protein has already been implicated in the self-aggregation of this protein. Thus fibrillar structure formation, nuclear disintegration and intermolecular p10 protein interactions seem to be three separate functions of the p10 protein and these functions are located in distinct domains of the protein. The mutants expressing truncated p10 proteins were impaired in electron-dense spacer formation but polyhedron envelopes were still formed. This result suggested that the formation of electron-dense spacers is not a prerequisite for the formation of polyhedron envelopes.

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© Society for General Microbiology 1993
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1993-04-01
2024-04-25
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References

  1. Bonner W. M. 1975; Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins. Journal of Cell Biology 64:421–430
     [Google Scholar]
  2. Breeuwer M., Goldfarb D. S. 1990; Facilitated transport of histone HI and other small nucleophilic proteins. Cell 60:999–1008
     [Google Scholar]
  3. Brown M., Faulkner P. 1977; A plaque assay for nuclear polyhedrosis viruses using a solid overlay. Journal of General Virology 36:361–364
     [Google Scholar]
  4. Cheley S., Kosik K. S., Paskevich P., Bakalis S., Bayley H. 1992; Phosphorylated baculovirus p 10 is a heat-stable microtubule-associated protein associated with process formation in Sf9 cells. Journal of Cell Science 102:739–752
     [Google Scholar]
  5. Clever J., Kasamatsu H. 1991; Simian virus 40 Vp2/3 small structural proteins harbor their own nuclear transport signal. Virology 181:78–90
     [Google Scholar]
  6. Emini E. A., Hughes J. V., Perlow D. S., Boger J. 1985; Induction of hepatitis A virus-neutralizing antibody by a virus specific synthetic peptide. Journal of Virology 55:836–839
     [Google Scholar]
  7. Flipsen J. TM., vanLent J. WM., Goldbach R. W., Vlak J. M. 1992; Induction of hepatitis A virus-neutralizing antibody by a virus specific synExpression of polyhedrin and p10 in the midgut of 4cMNPV infected Spodoptera exigua larvae: an immunoelectron microscopic investigation. JouJournal of Invertebrate Pathologyrnal of Virology 61:17–23
     [Google Scholar]
  8. Francki R. IB., Fauquet C. M., Knudson D. L., Brown F. 1991; Classification and nomenclature of viruses. Fifth Report of the International Committee on Taxonomy of Viruses. Archives of Virology supplementum 2:
     [Google Scholar]
  9. Gombart A. F., Pearson M. N., Rohrmann G. F., Beaudreau G. S. 1989; A baculovirus polyhedral envelope-associated protein: genetic location, nucleotide sequence, and immunocytochemical characterization. Virology 169:182–193
     [Google Scholar]
  10. Groebe D. R., Chung A. E., Ho C. 1990; Cationic lipid-mediated co-transfection of insect cells. Nucleic Acids Research 18:4033
     [Google Scholar]
  11. Hink F. 1970; Established insect cell line from the cabbage looper, Trichoplusia ni. Nature London 226:446–467
     [Google Scholar]
  12. Jarvis D. L., Bohlmeyer D. A., Garcia A. 1991; Requirements for nuclear localization and supramolecular assembly of a baculovirus polyhedrin protein. Virology 185:795–818
     [Google Scholar]
  13. Kuzio J., Rohel D. Z., Curry C. J., Krebs A., Carstens E. H., Faulkner P. 1984; Nucleotide sequence of the p10 polypeptide gene of Autographa californica nuclear polyhedrosis virus. Virology 139:414–418
     [Google Scholar]
  14. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydrophobic character of a protein. Journal of Molecular Biology 157:105–132
     [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 221:680–685
     [Google Scholar]
  16. Leisy D. J., Rohrmann G. F., Nesson M., Beaudreau G. S. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 221:680–685
     [Google Scholar]
  17. Leisy D. J., Rohrmann G. F., Nesson M., Beaudreau G. S. 1986; Nucleotide sequencing and transcriptional mapping of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus p10 gene. Virology 163:157–167
     [Google Scholar]
  18. MacKinnon E. A., Henderson J. F., Stolz D. B., Faulkner P. 1973; Morphogenesis of nuclear polyhedrosis virus under conditions of prolonged passage in vitro. Journal of Ultrastructure Research 49:419–435
     [Google Scholar]
  19. Pugsley A. P. 1989 The targeting of nuclear proteins. In Protein Targeting pp 199–210 San Diego: Academic Press;
     [Google Scholar]
  20. Quant-Russell R. L., Rohrmann G. F. 1990; A baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra. Virology 174:177–184
     [Google Scholar]
  21. Quant-Russell R. L., Pearson M. N., Rohrmann G. F., Beaudreau G. S. 1987; Characterization of baculovirus p10 synthesis using monoclonal antibodies. Virology 160:9–19
     [Google Scholar]
  22. Quant-Russell R. L., Pearson M. N., Rohrmann G. F. 1991; Immunoelectron microsopic examination of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected Lymantria dispar cells: time course and localization of major polyhedron-associated proteins. Journal of General Virology 72:275–283
     [Google Scholar]
  23. Rankin C., Ladin B. F., Weaver G. F. 1986; Physical mapping of temporally regulated overlapping transcripts in the region of the 10K protein of Autographa californica nuclear polyhedrosis virus. Journal of General Virology 57:18–27
     [Google Scholar]
  24. Roelvink P. W., van Meer M. MM., De Kort C. AD., Possee R. D., Hammock B. D., Vlak J. M. 1992; Dissimilar expression of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes. Journal of General Virology 73:1481–1489
     [Google Scholar]
  25. Rohel D. Z., Cochran M. A., Faulkner P. 1993; Characterization of two abundant mRNAs of Autographa californica nuclear polyhedrosis virus present late in infection. Virology 124:357–365
     [Google Scholar]
  26. Rohrmann G. F. 1986; Polyhedrin structure. Journal of General Virology 67:1499–1513
     [Google Scholar]
  27. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory Press;
     [Google Scholar]
  28. Smith G. E., Summers M. D. 1978; Analysis of baculovirus genomes with restriction enzyme endonucleases. Virology 98:517–527
     [Google Scholar]
  29. Smith G. E., Vlak J. M., Summers M. D. 1982; In vitro translation of Autographa californica nuclear polyhedrosis virus early and late mRNAs. Journal of Virology 44:199–208
     [Google Scholar]
  30. Smith G. E., Vlak J., Summers M. D. 1983; Physical analysis of Autographa californica nuclear polyhedrosis virus transcripts for polyhedrin and 10,000-molecular-weight protein. Journal of Virology 45:215–225
     [Google Scholar]
  31. van der Wilk F., vanLent J. WM., Vlak J. 1987; Immunogold detection of polyhedrin, p10 and virion antigens in Autographa californica nuclear polyhedrosis virus-infected Spodoptera frugiperda cells. Journal of General Virology 68:2615–2623
     [Google Scholar]
  32. vanLent J. WM., Groenen J. TM., Klinge-Roode E. C., Rohrmann G. E., Zuidema D., Vlak J. M. 1990; Localization of the 34 kDa polyhedron envelope protein in Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus. Virology 111:103–114
     [Google Scholar]
  33. van Oers M. M., Kormelink R. JM., Zuidema D., Vlak J. M. 1990; Functional studies on the p10 protein of Autographa californica nuclear polyhedrosis virus. Abstracts, VIIIth International Congress of VirologyP4–007
     [Google Scholar]
  34. van Strien E. A., Zuidema D., Goldbach R. W., Vlak J. M. 1992; Nucleotide sequence and transcriptional analysis of the polyhedrin gene of Spodoptera exigua nuclear polyhedrosis virus. Journal of General Virology 73:2813–2821
     [Google Scholar]
  35. 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]
  36. Vlak J. M., Klinkenberg F. A., Zaal K. GM., Usmany M., Klinge-Roode E. C., Geervuet J. BF., Roosien J., vanLent J. WM. 1988; Functional studies on the p10 gene of Autographa californica nuclear polyhedrosis virus using a recombinant expressing a p10-β-galactosidase fusion gene. Journal of General Virology 69:765–776
     [Google Scholar]
  37. Vlak J. M., Schouten A., Usmany M., Belsham G. J., Klinge-Roode E. C., Maule A. J., vanLent J. WM., Zuidema D. 1990; Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method. Virology 179:312–320
     [Google Scholar]
  38. Volkman L. E., Zaal K. JM. 1990; Autographa californica M nuclear polyhedrosis virus: microtubules and replication. Virology 175:292–302
     [Google Scholar]
  39. Whitt M. A., Manning J. S. 1988; A phosphorylated 34-kDa protein and a subpopulation of polyhedrin are thiol linked to the carbohydrate layer surrounding a baculovirus occlusion body. Virology 163:33–12
     [Google Scholar]
  40. Williams G. V., Rohel D. Z., Kuzio J., Faulkner P. 1989; A cytopathological investigation of Autographa californica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants. Journal of General Virology 70:187–202
     [Google Scholar]
  41. Wu J., Daugulis A. J., Faulkner P., Goosen M. F. 1992; Correlation of LDH activity with loss of insect cell viability: an assessment of LDH assay. Biotechnology Techniques 6:335–340
     [Google Scholar]
  42. Zuidema D., Klinge-Roode E. C., vanLent J. WM., Vlak J. M. 1989; Construction and analysis of an Autographa californica nuclear polyhedrosis virus mutant lacking the polyhedral envelope. Virology 173:98–108
     [Google Scholar]
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Functional domains of the p10 protein of Autographa californica nuclear polyhedrosis virus
J. Gen. Virol. 74, 563 (1993); https://doi.org/10.1099/0022-1317-74-4-563
/content/journal/jgv/10.1099/0022-1317-74-4-563
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