1887

Abstract

Serial passage of nuclear polyhedrosis viruses (NPVs) through cultured cell lines results in the appearance of mutants with a complex phenotype referred to as the ‘few polyhedra’ (FP) phenotype. The altered plaque morphology and reduced occlusion production associated with the FP phenotype have been observed in multinucleocapsid nuclear polyhedrosis virus (AcMNPV) bearing mutations in the gene encoding the 25 kDa protein (25K gene). In this study, we sequenced the 25K genes of four spontaneously occurring AcMNPV FP mutants. These mutants, together with an artificially generated FP mutant (AcFPβgal, in which the gene for β-galactosidase is fused in frame with the 25K ORF), were examined at the ultrastructural level to see if they exhibited the reduced virion occlusion and intranuclear envelopment which is associated with the FP phenotype. Observations on Sf9 cells infected with the FP mutants revealed that all five mutants were impaired in virion occlusion and intranuclear nucleocapsid envelopment. The 25K mutants were also found to release two- to fivefold more infectious virus (p.f.u.) into the media of infected Sf9 cells. Marker rescue of AcFPβgal restored wild-type virion occlusion, intranuclear envelopment and levels of infectious virus production.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-6-1451
1995-06-01
2022-05-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/6/JV0760061451.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-6-1451&mimeType=html&fmt=ahah

References

  1. Adams J. R., McClintock J. T. 1991; Baculoviridae. Nuclear Polyhedrosis Viruses, part 1: Nuclear Polyhedrosis Viruses of Insects. In Atlas of Invertebrate Viruses pp 87–204 Edited by Adams J. R., Bonami J. R. Boca Raton: CRC Press;
    [Google Scholar]
  2. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D. 1994; The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202:586–605
    [Google Scholar]
  3. Beames B., Summers M. D. 1988; Comparisons of host cell DNA insertions and altered transcription at the site of insertions in few polyhedra baculovirus mutants. Virology 162:206–220
    [Google Scholar]
  4. Beames B., Summers M. D. 1989; Location and nucleotide sequence of the 25 K protein missing from baculovirus few polyhedra (FP) mutants. Virology 168:344–353
    [Google Scholar]
  5. Bischoff D. S., Flaherty K. M., Hansel D., Mercer M. J., Kelly M. E., Hayes-Piazolles N. L., Slavicek J. M. 1994; Identification of five genomic loci in the baculovirus LdMNPV that impact on polyhedra formation. American Society for Virology 12 th Annual Meeting July 9–13 Madison, Wis., USA:
    [Google Scholar]
  6. Bishop D. H. L. 1992; Baculovirus expression vectors. Seminars in Virology 3:253–264
    [Google Scholar]
  7. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
    [Google Scholar]
  8. Couch J. A. 1991; Baculoviridae. Nuclear Polyhedrosis Viruses, part 2: Nuclear Polyhedrosis Viruses of Invertebrates other than Insects. In Atlas of Invertebrate Viruses pp 205–226 Edited by Adams J. R., Bonami J. R. Boca Raton: CRC Press;
    [Google Scholar]
  9. Fraser M.J. 1986; Transposon-mediated mutagenesis of baculo-viruses: transposon shuttling and implications for speciation. Annals of the Entomological Society of America 79:773–783
    [Google Scholar]
  10. Fraser M. J. 1987; FP mutation of nuclear polyhedrosis viruses: a novel system for the study of transposon-mediated mutagenesis. In Biotechnology in Invertebrate Pathology and Cell Culture pp 265–293 Edited by Maramorosch K. New York: Academic Press;
    [Google Scholar]
  11. Fraser M. J., Hink W. F. 1982; The isolation and characterization of the MP and FP plaque variants of Galleria mellonella polyhedrosis virus. Virology 117:366–378
    [Google Scholar]
  12. Fraser M. J., Smith G. E., Summers M. D. 1983; Acquisition of host cell DNA sequences by baculoviruses: relationship between host DNA insertions and FP mutants of Autographa californica and Galleria mellonella nuclear polyhedrosis viruses. Journal of Virology 47:287–300
    [Google Scholar]
  13. Fraser M. J., Brusca J. S., Smith G. E., Summers M. D. 1985; Transposon-mediated mutagenesis of a baculovirus. Virology 145:356–361
    [Google Scholar]
  14. Harrison R. L., Summers M. D. 1995; Biosynthesis and localization of the Autographa californica nuclear polyhedrosis virus 25K gene product. Virology (in press)
    [Google Scholar]
  15. Hink W. F., Vail P. V. 1973; A plaque assay for titration of alfalfa looper nuclear polyhedrosis virus in a cabbage looper (TN-368) cell line. Journal of Invertebrate Pathology 22:168–174
    [Google Scholar]
  16. Hink W. F., Strauss E. 1976; Replication and passage of alfalfa looper nuclear polyhedrosis virus plaque variants in cloned cell cultures and larval stages of four host species. Journal of Invertabrate Pathology 27:49–55
    [Google Scholar]
  17. Jarvis D. L., Bohlmeyer D. A., Garcia A. >Jr 1992; Enhancement of polyhedrin nuclear localization during baculovirus infection. Journal of Virology 66:6903–6911
    [Google Scholar]
  18. Jarvis D. L., Summers M. D., Garcia A. Jr, Bohlmeyer D. A. 1993; Influence of different signal peptides and prosequences on expression and secretion of human tissue plasminogen activator in the baculovirus system. Journal of Biological Chemistry 268:16754–16762
    [Google Scholar]
  19. Knudson D. L., Harrap K. A. 1976; Replication of a nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda \ microscopy study of the sequence of events of the virus infection. Journal of Virology 17:254–268
    [Google Scholar]
  20. Kumar S., Miller L. K. 1987; Effects of serial passage of Autographa californica nuclear polyhedrosis virus in cell culture. Virus Research 7:335–349
    [Google Scholar]
  21. Luckow V. A., Summers M. D. 1988a; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  22. Luckow V. A., Summers M. D. 1988b; Signals important for high-level expression of foreign genes m Autographa californica nuclear polyhedrosis virus expression vectors. Virology 167:56–71
    [Google Scholar]
  23. Mackinnon E. A., Henderson J. F., Stoltz D. B., Faulkner P. 1974; Morphogenesis of nuclear polyhedrosis virus under conditions of prolonged passage in vitro. Journal of Ultrastructure Research 49:419–435
    [Google Scholar]
  24. Murhammer D. W., Goochee C. F. 1988; The scaleup of insect cell cultures: protective effects of pluronic F-68. Bio/Technology 6:1411–1418
    [Google Scholar]
  25. Potter K. N., Faulkner P., MacKinnon E. A. 1976; Strain selection during serial passage of Trichoplusia m nuclear polyhedrosis virus. Journal of Virology 18:1040–1050
    [Google Scholar]
  26. Ramoska W. A., Hink W. F. 1974; Electron microscope examination of two plaque variants from a nuclear polyhedrosis virus of the alfalfa looper, Autographacalifornica. Journal of Invertebrate Pathology 23:197–201
    [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences 74:5463–5467
    [Google Scholar]
  28. Slavicek J. M., Podgewaite J., Lanner-Herrera C. 1992; Properties of two Lymantra dispar nuclear polyhedrosis virus isolates obtained from the microbial pesticide Gypchek. Journal of Invertebrate Pathology 59:142–148
    [Google Scholar]
  29. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures Texas Agricultural Experimental Station Bulletin number 1555.;
    [Google Scholar]
  30. 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]
  31. Venable J. H., Coggeshall R. 1965; A simplified lead citrate stain for use in electron microscopy. Journal of Cell Biology 25:407–108
    [Google Scholar]
  32. Wang H. H., Fraser M.J., Cary L. C. 1989; Transposon mutagenesis of baculoviruses: analysis of TFP3 lepidopteran transposon insertions at the FP locus of nuclear polyhedrosis viruses. Gene 81:97–108
    [Google Scholar]
  33. Wood H. A. 1980; Isolation and replication of an occlusion body-deficient mutant of the Autographa californica nuclear polyhedrosis vims. Virology 105:338–344
    [Google Scholar]
  34. Wood H. A., Granados R. R. 1991; Genetically engineered baculoviruses as agents for pest control. Annual Review of Microbiology 45:69–87
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-6-1451
Loading
/content/journal/jgv/10.1099/0022-1317-76-6-1451
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