Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1997 Oct;71(10):7593–7599. doi: 10.1128/jvi.71.10.7593-7599.1997

Expression of the Autographa californica nuclear polyhedrosis virus apoptotic suppressor gene p35 in nonpermissive Spodoptera littoralis cells.

E Gershburg 1, H Rivkin 1, N Chejanovsky 1
PMCID: PMC192107  PMID: 9311840

Abstract

Apoptosis was postulated as the main barrier to replication of the Autographa californica nuclear polyhedrosis virus (AcMNPV) in a Spodoptera littoralis SL2 cell line (N. Chejanovsky and E. Gershburg, Virology 209:519-525, 1995). Thus, we hypothesized that the viral apoptotic suppressor gene p35 is either poorly expressed or nonfunctional in AcMNPV-infected SL2 cells. These questions were addressed by first determining the steady-state levels of the p35 product, P35, in AcMNPV-infected SL2 cells. Indeed, very low levels of P35 were found in infected SL2 cells in comparison with those in SF9 cells. Overexpression of p35, in transient-transfection and recombinant-virus infection experiments, inhibited actinomycin D- and AcMNPV-induced apoptosis, as determined by reduced cell blebbing and release of oligonucleosomes and increased cell viability of SL2. However, SL2 budded-virus (BV) titers of a recombinant AcMNPV which highly expressed p35 did not improve significantly. Also, injection of S. littoralis larvae with recombinant and wild-type AcMNPV BVs showed similar 50% lethal doses. These data suggest that apoptosis is not the only impediment to AcMNPV replication in these nonpermissive S. littoralis cells, and probably in S. littoralis larvae, so p35 may not be the only host range determinant in this system.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D. The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology. 1994 Aug 1;202(2):586–605. doi: 10.1006/viro.1994.1380. [DOI] [PubMed] [Google Scholar]
  2. Bertin J., Mendrysa S. M., LaCount D. J., Gaur S., Krebs J. F., Armstrong R. C., Tomaselli K. J., Friesen P. D. Apoptotic suppression by baculovirus P35 involves cleavage by and inhibition of a virus-induced CED-3/ICE-like protease. J Virol. 1996 Sep;70(9):6251–6259. doi: 10.1128/jvi.70.9.6251-6259.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnbaum M. J., Clem R. J., Miller L. K. An apoptosis-inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with Cys/His sequence motifs. J Virol. 1994 Apr;68(4):2521–2528. doi: 10.1128/jvi.68.4.2521-2528.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blissard G. W., Rohrmann G. F. Baculovirus diversity and molecular biology. Annu Rev Entomol. 1990;35:127–155. doi: 10.1146/annurev.en.35.010190.001015. [DOI] [PubMed] [Google Scholar]
  5. Bonning B. C., Hammock B. D. Development and potential of genetically engineered viral insecticides. Biotechnol Genet Eng Rev. 1992;10:455–489. doi: 10.1080/02648725.1992.10647895. [DOI] [PubMed] [Google Scholar]
  6. Bonning B. C., Hammock B. D. Development of recombinant baculoviruses for insect control. Annu Rev Entomol. 1996;41:191–210. doi: 10.1146/annurev.en.41.010196.001203. [DOI] [PubMed] [Google Scholar]
  7. Bump N. J., Hackett M., Hugunin M., Seshagiri S., Brady K., Chen P., Ferenz C., Franklin S., Ghayur T., Li P. Inhibition of ICE family proteases by baculovirus antiapoptotic protein p35. Science. 1995 Sep 29;269(5232):1885–1888. doi: 10.1126/science.7569933. [DOI] [PubMed] [Google Scholar]
  8. Cartier J. L., Hershberger P. A., Friesen P. D. Suppression of apoptosis in insect cells stably transfected with baculovirus p35: dominant interference by N-terminal sequences p35(1-76). J Virol. 1994 Dec;68(12):7728–7737. doi: 10.1128/jvi.68.12.7728-7737.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chejanovsky N., Gershburg E. The wild-type Autographa californica nuclear polyhedrosis virus induces apoptosis of Spodoptera littoralis cells. Virology. 1995 Jun 1;209(2):519–525. doi: 10.1006/viro.1995.1284. [DOI] [PubMed] [Google Scholar]
  10. Chejanovsky N., Zilberberg N., Rivkin H., Zlotkin E., Gurevitz M. Functional expression of an alpha anti-insect scorpion neurotoxin in insect cells and lepidopterous larvae. FEBS Lett. 1995 Dec 4;376(3):181–184. doi: 10.1016/0014-5793(95)01263-4. [DOI] [PubMed] [Google Scholar]
  11. Clem R. J., Fechheimer M., Miller L. K. Prevention of apoptosis by a baculovirus gene during infection of insect cells. Science. 1991 Nov 29;254(5036):1388–1390. doi: 10.1126/science.1962198. [DOI] [PubMed] [Google Scholar]
  12. Clem R. J., Miller L. K. Apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. J Virol. 1993 Jul;67(7):3730–3738. doi: 10.1128/jvi.67.7.3730-3738.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Clem R. J., Miller L. K. Control of programmed cell death by the baculovirus genes p35 and iap. Mol Cell Biol. 1994 Aug;14(8):5212–5222. doi: 10.1128/mcb.14.8.5212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Croizier G., Croizier L., Argaud O., Poudevigne D. Extension of Autographa californica nuclear polyhedrosis virus host range by interspecific replacement of a short DNA sequence in the p143 helicase gene. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):48–52. doi: 10.1073/pnas.91.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Crook N. E., Clem R. J., Miller L. K. An apoptosis-inhibiting baculovirus gene with a zinc finger-like motif. J Virol. 1993 Apr;67(4):2168–2174. doi: 10.1128/jvi.67.4.2168-2174.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Du X., Thiem S. M. Characterization of host range factor 1 (hrf-1) expression in Lymantria dispar M nucleopolyhedrovirus- and recombinant Autographa californica M nucleopolyhedrovirus-infected IPLB-Ld652Y cells. Virology. 1997 Jan 20;227(2):420–430. doi: 10.1006/viro.1996.8356. [DOI] [PubMed] [Google Scholar]
  17. Grether M. E., Abrams J. M., Agapite J., White K., Steller H. The head involution defective gene of Drosophila melanogaster functions in programmed cell death. Genes Dev. 1995 Jul 15;9(14):1694–1708. doi: 10.1101/gad.9.14.1694. [DOI] [PubMed] [Google Scholar]
  18. Guarino L. A., Summers M. D. Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. J Virol. 1986 Feb;57(2):563–571. doi: 10.1128/jvi.57.2.563-571.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hay B. A., Wolff T., Rubin G. M. Expression of baculovirus P35 prevents cell death in Drosophila. Development. 1994 Aug;120(8):2121–2129. doi: 10.1242/dev.120.8.2121. [DOI] [PubMed] [Google Scholar]
  20. Hershberger P. A., Dickson J. A., Friesen P. D. Site-specific mutagenesis of the 35-kilodalton protein gene encoded by Autographa californica nuclear polyhedrosis virus: cell line-specific effects on virus replication. J Virol. 1992 Sep;66(9):5525–5533. doi: 10.1128/jvi.66.9.5525-5533.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hershberger P. A., LaCount D. J., Friesen P. D. The apoptotic suppressor P35 is required early during baculovirus replication and is targeted to the cytosol of infected cells. J Virol. 1994 Jun;68(6):3467–3477. doi: 10.1128/jvi.68.6.3467-3477.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kool M., Ahrens C. H., Vlak J. M., Rohrmann G. F. Replication of baculovirus DNA. J Gen Virol. 1995 Sep;76(Pt 9):2103–2118. doi: 10.1099/0022-1317-76-9-2103. [DOI] [PubMed] [Google Scholar]
  23. LaCount D. J., Friesen P. D. Role of early and late replication events in induction of apoptosis by baculoviruses. J Virol. 1997 Feb;71(2):1530–1537. doi: 10.1128/jvi.71.2.1530-1537.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Lu A., Miller L. K. Species-specific effects of the hcf-1 gene on baculovirus virulence. J Virol. 1996 Aug;70(8):5123–5130. doi: 10.1128/jvi.70.8.5123-5130.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lu A., Miller L. K. The roles of eighteen baculovirus late expression factor genes in transcription and DNA replication. J Virol. 1995 Feb;69(2):975–982. doi: 10.1128/jvi.69.2.975-982.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maeda S., Kamita S. G., Kondo A. Host range expansion of Autographa californica nuclear polyhedrosis virus (NPV) following recombination of a 0.6-kilobase-pair DNA fragment originating from Bombyx mori NPV. J Virol. 1993 Oct;67(10):6234–6238. doi: 10.1128/jvi.67.10.6234-6238.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Martinou I., Fernandez P. A., Missotten M., White E., Allet B., Sadoul R., Martinou J. C. Viral proteins E1B19K and p35 protect sympathetic neurons from cell death induced by NGF deprivation. J Cell Biol. 1995 Jan;128(1-2):201–208. doi: 10.1083/jcb.128.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Miller L. K. Genetically engineered insect virus pesticides: present and future. J Invertebr Pathol. 1995 May;65(3):211–216. doi: 10.1006/jipa.1995.1032. [DOI] [PubMed] [Google Scholar]
  30. Palli S. R., Caputo G. F., Sohi S. S., Brownwright A. J., Ladd T. R., Cook B. J., Primavera M., Arif B. M., Retnakaran A. CfMNPV blocks AcMNPV-induced apoptosis in a continuous midgut cell line. Virology. 1996 Aug 1;222(1):201–213. doi: 10.1006/viro.1996.0411. [DOI] [PubMed] [Google Scholar]
  31. Rabizadeh S., LaCount D. J., Friesen P. D., Bredesen D. E. Expression of the baculovirus p35 gene inhibits mammalian neural cell death. J Neurochem. 1993 Dec;61(6):2318–2321. doi: 10.1111/j.1471-4159.1993.tb07477.x. [DOI] [PubMed] [Google Scholar]
  32. Sugimoto A., Friesen P. D., Rothman J. H. Baculovirus p35 prevents developmentally programmed cell death and rescues a ced-9 mutant in the nematode Caenorhabditis elegans. EMBO J. 1994 May 1;13(9):2023–2028. doi: 10.1002/j.1460-2075.1994.tb06475.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thiem S. M., Du X., Quentin M. E., Berner M. M. Identification of baculovirus gene that promotes Autographa californica nuclear polyhedrosis virus replication in a nonpermissive insect cell line. J Virol. 1996 Apr;70(4):2221–2229. doi: 10.1128/jvi.70.4.2221-2229.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Todd J. W., Passarelli A. L., Miller L. K. Eighteen baculovirus genes, including lef-11, p35, 39K, and p47, support late gene expression. J Virol. 1995 Feb;69(2):968–974. doi: 10.1128/jvi.69.2.968-974.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Xue D., Horvitz H. R. Inhibition of the Caenorhabditis elegans cell-death protease CED-3 by a CED-3 cleavage site in baculovirus p35 protein. Nature. 1995 Sep 21;377(6546):248–251. doi: 10.1038/377248a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES