1887

Abstract

Immunoelectron microscopy was employed to examine the temporal expression and localization of two proteins involved in baculovirus polyhedron assembly (polyhedrin and p10) of multicapsid nuclear polyhedrosis virus (OpMNPV) in infected cells. In addition, the association of p10 with the polyhedron envelope (PE) protein was studied. The major capsid protein (p39) was also examined to investigate the association of virion structural proteins with polyhedron formation. In infected cells, p39 did not show a concentrated association with any infected-cell structures other than nucleocapsids and appeared to be randomly distributed over the nucleocapsid surface. Likewise, polyhedrin showed no major concentrations outside of developing or mature polyhedra. The p10 antibody cross-reacted with a protein associated with condensed chromosomes in uninfected cells. In infected cells, p10 is a component of the body of fibrillar structures. The PE protein has been shown to accumulate around the periphery of fibrillar structures. Cells infected with a polyhedrin-minus virus expressing the β-galactosidase gene under the control of the polyhedrin promoter were examined to determine whether the lack of polyhedra would influence the localization of major polyhedron-associated viral proteins. High concentrations of PE protein accumulating on the periphery of fibrillar structures appeared to be the major difference from wild-type virus-infected cells. The β-galactosidase protein appeared to be distributed throughout the nucleus and cytoplasm, in contrast with the specific localization of the viral proteins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-2-275
1991-02-01
2022-01-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/2/JV0720020275.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-2-275&mimeType=html&fmt=ahah

References

  1. Bradford M. B., Blissard G. W., Rohrmann G. F. 1990; Characterization of the infection cycle of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus in Lymantria dispar cells. Journal of General Virology 71:2841–2846
    [Google Scholar]
  2. Fung M., Chiu K. Y. M., Weber T., Chang T., Chang N. T. 1988; Detection and purification of a recombinant human β lymphotropic virus (HHV-6) in the baculovirus expression system by limiting dilution and DNA dot-blot hybridization. Journal of Virological Methods 19:33–42
    [Google Scholar]
  3. 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]
  4. 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 153:157–167
    [Google Scholar]
  5. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  6. Miller J. H. 1972 Experiments in Molecular Genetics New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  7. Pearson M. N., Russell R. L. Q., Rohrmann G. F., Beaudreau G. S. 1988; P39, a major baculovirus structural protein: immuno-cytochemical characterization and genetic location. Virology 167:407–413
    [Google Scholar]
  8. Quant R. L., Pearson M. N., Rohrmann G. F., Beaudreau G. S. 1984; Production of polyhedrin monoclonal antibodies for distinguishing two Orgyia pseudotsugata baculoviruses. Applied and Environmental Microbiology 48:732–736
    [Google Scholar]
  9. 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]
  10. Russell R. L. Q., Rohrmann G. F. 1990; A baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra. Virology 174:177–184
    [Google Scholar]
  11. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures Texas Agricultural Experiment Station Bulletin, no. 1555
    [Google Scholar]
  12. Thiem S. M., Miller L. K. 1989; Identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus Autographa califomica nuclear polyhedrosis virus. Journal of Virology 63:2008–2018
    [Google Scholar]
  13. Van Der Wilk F., Van Lent J. W. M., Vlak J. M. 1987; Immunogold detection of polyhedrin, p10 and virion antigens in Autographa califomica nuclear polyhedrosis virus-infected Spodoptera frugiperda cells . Journal of General Virology 68:2615–2623
    [Google Scholar]
  14. van Lent J. W. M., Groenen J. T. M., Klinge-Roode E. C., Rohrmann G. F., Zuidema D., Vlak J. M. 1990; Localization of the 34-kDa polyhedron envelope protein in Spodoptera frugiperda cells infected with Autographa califomica nuclear polyhedrosis virus. Archives of Virology 111:103–114
    [Google Scholar]
  15. Vlak J. M., Klinkenberg F. A., Zaal K. J. M., Usmany M., Klinge-Roode E. C., Geervliet J. B. F., Roosien J., van Lent J. W. M. 1988; Functional studies on the p10 gene of Autographa califomica nuclear polyhedrosis virus using a recombinant expressing a p10–β- galactosidase fusion gene. Journal of General Virology 69:765–776
    [Google Scholar]
  16. Volkman L. E., Zaal K. J. M. 1990; Autographa califomica M nuclear polyhedrosis virus: microtubules and replication. Virology 175:292–302
    [Google Scholar]
  17. 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 163:33–42
    [Google Scholar]
  18. Williams G. V., Rohel D. Z., Kuzio J., Faulkner P. 1989; A cytopathological investigation of Autographa califomica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants. Journal of General Virology 70:187–202
    [Google Scholar]
  19. Zuidema D., Klinge-Roode E. C., van Lent J. W. M., Vlak J. M. 1989; Construction and analysis of an Autographa califomica nuclear polyhedrosis virus mutant lacking the polyhedral envelope. Virology 173:98–108
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-2-275
Loading
/content/journal/jgv/10.1099/0022-1317-72-2-275
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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