1887

Abstract

Baculovirus-mediated abnormal protein trafficking was studied in infected caterpillars by using heterologous proteins. The gene for human complement C1r was expressed in larvae of by a recombinant nuclear polyhedrosis virus (AcMNPV) vector. By following the time-course of recombinant C1r distribution among various tissues, cell types and cell organelles, we concluded that the dominant site of recombinant protein synthesis was the fat body, although some production in the haemocytes and midgut was also observed. Only about 4% of the cells of the infected organs expressed recombinant C1r, which was then secreted into the haemolymph. The tracheal and integumental cuticle was rich in recombinant protein from the fourth day after infection although epidermal cells did not synthesize recombinant C1r. The morphological picture suggested that the accumulation was a consequence of a trans-epithelial transport. This transport process was checked by following the fate of the 49 kDa haemolymph protein and injected ovalbumin in AcMNPV-infected and in nuclear polyhedrosis virus- infected larvae. Both proteins were able to pass the basal membrane of the epidermis and accumulated in the cuticle, while in control larvae neither was transported. The observed transepithelial transport points to the role of baculo- viruses in directing recombinant, endo- and exogenous proteins to cuticulated tissues. Based on these results we conclude that the permeability of basal membranes undergoes a characteristic change during the course of baculovirus infection.

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1997-04-01
2024-04-20
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References

  1. 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]
  2. Booth T. F., Bonning B. C., Hammock B. D. 1992; Localization of juvenile hormone esterase during development in normal and in recombinant baculovirus infected larvae of the moth Trichoplusia ni. Tissue & Cell 24:267–282
    [Google Scholar]
  3. Clem R. J., Robson M., Miller L. K. 1994; Influence of infection route on the infectivity of baculovirus mutants lacking the apoptosis- inhibiting gene p35 and the adjacent gene p94. Journal of Virology 68:6759–6762
    [Google Scholar]
  4. Engelhard E. K., Kam-Morgan L. N. W., Washburn J. O., Volkman L. E. 1994; The insect tracheal system: a conduit for the systemic spread of Autographa californica M nuclear polyhedrosis virus. Proceedings of the National Academy of Sciences, USA 91:3224–3227
    [Google Scholar]
  5. Flipsen J. T. M., Martens J. W. M., Van Oers M. M., Vlak J. M., VanLent J. W. M. 1995; Transition from primary to secondary infection of AcMNPV In the midgut of Spodoptera exigua larvae. Virology 208:328–335
    [Google Scholar]
  6. Gál P., Sárvári M., Szilágyi K., Závodszky P., Schumaker V. N. 1989; Expression of hemolytically active human complement Clr proenzyme in insect cells using a baculovirus vector. Complement and Inflammation 6:433–441
    [Google Scholar]
  7. Granados R. R., Lawler K. A. 1981; In vivo pathway of Autographa californica baculovirus invasion and infection. Virology 108:297–308
    [Google Scholar]
  8. Granados R. R., Williams K. 1986; In vivo infection and replication ofbaculoviruses. In The BiologyofBaculoviruses 1 pp. 89–108 Granados R., Federici B. Edited by Boca Raton, Fla: CRC Press;
    [Google Scholar]
  9. Harrap K., Longworth J. 1974; An evaluation of purification methods for baculoviruses. Journal of Invertebrate Pathology 24:55–62
    [Google Scholar]
  10. Herbert W. J. 1978; Laboratory animal techniques for immunology. In HandbookofExperimentalImmunology Appendix 4 Weir D. M. Edited by London: Blackwell Scientific Publications;
    [Google Scholar]
  11. Hunter F. R., Crook N. E., Entwistle P. F. 1984; Viruses as pathogens for the control of insects. In Microbial Methods for Environmental Biotechnology pp. 323–347 Grainger J. M., Lynch J. M. Edited by New York & London: Academic Press;
    [Google Scholar]
  12. Keddie B. A., Aponte G. W., Volkman L. E. 1989; The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science 243:1728–1730
    [Google Scholar]
  13. Kool M., Ahrens C. H., Vlak J. M., Rohrmann G. F. 1995; Replication of baculovirus DNA. Journal of General Virology 76:2103–2118
    [Google Scholar]
  14. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  15. Leung H., Palli S. R., Locke M. 1989; The localization of arylphorin in an insect, Calpodes ethlius. Journal of Insect Physiology 35:223–231
    [Google Scholar]
  16. Leytus S. P., Kurachi K., Sakariassen K. S., Davie E. W. 1986; Nucleotide sequence of the cDNA coding for human complement C1r. Biochemistry 25:4855–4863
    [Google Scholar]
  17. Locke M. 1994; Preservation and contrast without osmication or section staining. Microscopy Research and Techniques 29:1–10
    [Google Scholar]
  18. Maeda S., Kawai T., Obinata M., Fujiwara H., Horiuchi T., Saeki Y., Sato Y., Furusawa M. 1985; Production of human alpha-interferon in silkworm using a baculovirus vector. Nature 315:592–594
    [Google Scholar]
  19. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors : the requirements for high-level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  20. O’Reilly D. R., Miller L. K., Luckow V. A. 1992 Baculovirus Expression Vectors New York: W. H. Freeman;
    [Google Scholar]
  21. Pennock G. D., Shoemaker C., Miller L. K. 1984; Strong and regulated expression of Escherichia coli β-galactosidase in insect cells with a baculovirus vector. Molecular and Cellular Biology 4:399–406
    [Google Scholar]
  22. Sass M., Kiss A., Locke M. 1993; Classes of integument peptides. Insect Biochemistry and Molecular Biology 23:845–857
    [Google Scholar]
  23. Sárvári M., Csikós Gy., Sass M., Gál P., Schumaker V. N., Závodszky P. 1990; Ecdysteroids increase the yield of recombinant protein produced in baculovirus insect cell expression system. Biochemical and Biophysical Research Communications 167:1154–1161
    [Google Scholar]
  24. Smith G. E., Fraser M. J., Summers M. D. 1983; Molecular engineering of Autographa californica nuclear polyhedrosis virus genome: deletion mutations within the polyhedrin gene. Journal of Virology 46:584–593
    [Google Scholar]
  25. Smith I. R. L., Van Beek N. A. M., Podgwaite J. D., Wood H. A. 1988; Physical map and polyhedrin gene sequence of Lymantria dispar nuclear polyhedrosis virus. Gene 71:97–105
    [Google Scholar]
  26. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures Texas Agricultural Experiment Station Bulletin 1555
    [Google Scholar]
  27. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proceedings of the National Academy of Sciences, USA 76:4350–4354
    [Google Scholar]
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