The Homologous Region 3 from Bombyx mori Nucleopolyhedrovirus Enhancing the Transcriptional Activity of Drosophila hsp70 Promoter

  • Tang, Shun-Ming (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute) ;
  • Yi, Yong-Zhu (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute) ;
  • Zhou, Ya-Jing (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute) ;
  • Zhang, Zhi-Fang (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute) ;
  • Li, Yi-Ren (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute) ;
  • He, Jia-Lu (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute)
  • Published : 2004.12.01

Abstract

Drosophila melanogaster heat shock protein 70 gene promoter (Dhsp70p) is widely used in transgenic insect to drive exogenous gene, and the homologous region 3 from Bombyx mori nucleopolyhedrovirus (BmNPVhr3) functions as an enhancer for several promoters. To test whether BmNPVhr3 can enhance the Dhsp70ps transcriptional activity, the reporter plasmids, which contain the Dhsp70p, the reporter $\beta$-galactosidase gene with SV40 terminator and BmNPVhr3 fragment, are constructed and transfected into the insect cell lines (Bm-N cells and Sf-21 cells) by lipofectin-mediated method. The results from the transient expression assay show that BmNPVhr3 significantly increases transcriptional activity of Dhsp70p both under the normal condition and under the heat-shock treatment, although the effects are significantly different between in Bm-N cells and in sf-21 cells. The enhancing behavior of BmNPVhr3 on the Dhsp70p is in an orientation-independent manner. Meanwhile, the effects of heat-shock treatment on Dhsp70p alone or Dhsp70p/BmNPVhr3 combination present no significant difference, indicating that BmNPVhr3 only enhances the transcriptional activity of Dhsp70p, but cant alter its characteristic of the response to the heat-shock stress. The above results suggest that the Dhsp70p/BmNPVhr3 combination is more effective one to drive exogenous gene for transgene or stable cell expression system in insects.

Keywords

References

  1. Beckmann, R. P., L. A. Mizzen and W. J. Weilch (1990) Interaction of hsp70 with newly synthesized proteins: implication for protein folding and assembly. Science 248, 850-853 https://doi.org/10.1126/science.2188360
  2. Cotto, J. J. and R. I. Morimoto (1999) Stress-induced activation of the heat-shock response: cell and molecular biology of heat-shock factors. Biochem. Soc. Symp. 64,105-118
  3. Feder, M. E. and G. E. Hofmann (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu. Rev. Physiol. 61, 243-282 https://doi.org/10.1146/annurev.physiol.61.1.243
  4. Gomi, S., K. Majima and S. Maeda (1999) Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J. Gen. Virol. 80, 1323-1337 https://doi.org/10.1099/0022-1317-80-5-1323
  5. Hung, J. J., T. J. Cheng, M. D. Chang, K. D. Chen, H. L. Huang and Y. K. Lai (1998) Involvement of heat shock elements and basal transcription elements in the differential induction of the 70-kDa heat shock protein and its cognate by cadmium chloride in 9L rat brain tumor cells. J. Cell Biochem. 71, 21-35 https://doi.org/10.1002/(SICI)1097-4644(19981001)71:1<21::AID-JCB3>3.0.CO;2-3
  6. Huynh, C. Q. and H. Zieler (1999) Construction of modular and versatile plasmid vectors for the high-level expression of single or multiple genes in insects and insect cell lines. J. Mol. BioI. 288, 13-20 https://doi.org/10.1006/jmbi.1999.2674
  7. Idahl, L. A., P. E. Sandatom and J. Sehlin (1986) Measurements of serum glucose using the luciferin/luciferase system and a liquid scintillation spectrometer. Analyt. Biochem. 155, 177-181 https://doi.org/10.1016/0003-2697(86)90243-5
  8. Jeffrey, H. M. (1992) Experiments in Molecular Genetics, 11th printing, Cold Spring Harbor Laboratory Press, New York, pp. 398-404
  9. Kimura, R. H., P. V. Choudary and C. W. Schmid (1999) Silkworm Bm1 SINE RNA increases following cellular insults. Nucleic Acids Res. 27, 3380-3387 https://doi.org/10.1093/nar/27.16.3380
  10. Lacoste, A., M. C. De Cian, A. Cueff and S. A. Poulet (2001) Noradrenaline and alpha-adrenergic signaling induce the hsp70 gene promoter in mollusc immune cells. J. Cell Sci. 114, 3557-3564
  11. Lei, X. D., Y. D. Mi, Z. Y. Yuan, Z. P. Li and X. F. Wu (1994) Expression of luciferase gene in silkworm. Chin. Sci. Bull. 39, 847-849
  12. Lindquist, S. and E. A. Craig (1988) The heat-shock proteins. Annu. Rev. Genet. 22, 631-677 https://doi.org/10.1146/annurev.ge.22.120188.003215
  13. Lo, H. R., C. C. Chou, T. Y. Wu, J. P. Yuen and Y. C. Chao (2002) Novel baculovirus DNA elements strongly stimulate activities of exogenous and endogenous promoters. J. Biol. Chem. 277, 5256-5264 https://doi.org/10.1074/jbc.M108895200
  14. Lu, M., P. J. Farrell, R. Johnson and K. latrou (1997) A baculovirus (Bombyx mori nuclear polyhedrosis virus) repeat element functions as a powerful constitutive enhancer in transfected insect cells. J. Bioi. Chem. 272, 30724-30728 https://doi.org/10.1074/jbc.272.49.30724
  15. Moos, M. (1995) Analysis of proteins; in Short Protocols in Molecular Biology. Ausubel, F. M., R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl (eds.), pp. 10.1.4-10.1.5. John Wiley & Sons, Inc
  16. Ricchi, A., G. Cardu, B. Lettieri, D. E. Fusar-Poli, C. Tacchini, A. Bernelli-Zazzera and G. G. Corbucci (2001) Biomolecular and biochemical response of myocardial cell to ischemia and reperfusion in the course of heart surgery. J. Cardiovasc. Surg. 42, 605-610
  17. Sambrook, J., E. F. Fritsch and T. Maniatis (1989) Molecular Cloning: A Laboratory Manual. 2nd ed, Cold Spring Harbor Laboratory Press, New York
  18. Schmidt, M., T. Heimberger, P. Gruensfelder, G. Schler and F. Hoppe (2004) Inducible promoters for gene therapy of head and neck cancer: an in vitro study. Eur. Arch. Otorhinolaryngol. 261, 208-215 https://doi.org/10.1007/s00405-003-0621-z
  19. Shen, X. J., Y. Z. Yi, S. M. Tang, Z. F. Zhang, Y. R. Li, J. L. He and X. F. Wu (2004) Characterization of ecdysteroid UDP-glucosyltransferase gene promoter from Bombyx mori nucleopolyhedrovirus. Int. J. Indust. Entomol. 8, 169-174
  20. Statistical Analysis System Users Guide (1990) Version 6.12, SAS. Institute Inc. Cary. North Carolina: Institute Inc. Cary. North Carolina, Vols. 1.2 (on disc)
  21. Summers, M. D. and G. E. Smith (1987) A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin, No. 1555
  22. Tang, S. M., Y. Z. Yi, X. J. Shen, Z. F. Zhang, Y. R. Li and J. L. He (2003) Functional analysis of the larval serum protein gene promoter from silkworm, Bombyx mori. Chin. Sci. Bull. 48, 2611-2615 https://doi.org/10.1360/03wc0444
  23. Uhlirova, M., M. Asahina, L. M. Riddiford and M. Jindra (2002) Heat-inducible transgenic expression in the silkmoth Bombyx mori. Dev. Genes Evol. 212, 145-151 https://doi.org/10.1007/s00427-002-0221-8
  24. Viswanathan, P., B. Venkaiah, M. S. Kumar, S. Rasheedi, S. Vrati, M. D. Bashyam and S. E. Hasnain (2003) The homologous region sequence (hr1) of Autographa californica multinucleocapsid polyhedrosis virus can enhance transcription from non-baculoviral promoters in mammalian cells. J. BioI. Chem. 278, 52564-52571 https://doi.org/10.1074/jbc.M309351200
  25. Xiao, Q. L., Z. F. Zhang, Y. Z. Yi, J. L. He and X. F. Wu (2001) Functional analysis of helicase gene promoter and homologous region 3 enhancer in Bombyx mori nuclear polyhedrosis virus. Acta Biochim. Biophys. Sin. 33, 525-530
  26. Yanagida, Y., A. Mizuno, T. Motegi, E. Kobatake and M. Aizawa (2000) Electrically stimulated induction of hsp70 gene expression in mouse astroglia and fibroblast cells. J. Biotechnol. 79, 53-61 https://doi.org/10.1016/S0168-1656(00)00214-5
  27. Zhang, Z. F., Y. Zhang, H. S. Lu and X. F. Wu (1995) Structure and function of DNA replication origin hr3 of Bombyx mori nuclear polyhedrosis virus. Sci. China Series B. 25, 949-955
  28. Zhao, Y. G. and P. Eggleston (1999) Comparative analysis of promoters for transient gene expression in cultured mosquito cells. Insect Mol. Biol. 8, 31-38 https://doi.org/10.1046/j.1365-2583.1999.810031.x
  29. Zhou, Y. J. (2002) Effects of insect hormones and CTAB on the baculovirus replication and related gene promoter activity. Ph. D. Thesis, Engineering University of East China
  30. Zhou, Y. J., Y. Z. Yi, Z. F. Zhang, J. L. He, Y. X. Zhang and X. F.Wu (2003) Promoter activities in the baculovirus envelope glycoprotein gp64 gene. Acta Biochim. Biophys. Sin. 35, 18-26
  31. Zuidema, D., A. Schouten, M. Usmany, A. J. Maule, G. J. Belsham, J. Roosien, E. C. Klinge-Roode, J. W. van Lent and J. M. Vlak (1990) Expression of cauliflower mosaic virus gene I in insect cells using a novel polyhedrin-based baculovirus expression vector. J. Gen. Virol. 71, 2201-2209 https://doi.org/10.1099/0022-1317-71-10-2201