Stimulation of baculovirus transcriptome expression in mammalian cells by baculoviral transcriptional activators Free

Abstract

(AcMNPV), the type species of the family , is an insect-specific virus that can enter a variety of mammalian cells. The potential of this versatile virus for protein expression or gene therapy in mammalian cells has become the focus of many studies. In most mammalian cells, transduced AcMNPV genes are either not expressed or expressed at an extremely low level. Here, we studied the effects of the two major AcMNPV -activators, IE1 and IE2, on the activation of AcMNPV genome in Vero E6 cells. Microarray analysis showed that when IE1 was overexpressed, it significantly activated genes and , and upregulated , , , , and . Although, there were only two genes, and , that were activated by IE2, we discovered interestingly that the combination of IE1 and IE2 factors had a synergistic effect on activation of the AcMNPV genome in mammalian cells, and activated around 38 %, or 59 out of the 155 genes placed on the microarray. This is the first detailed study of baculoviral transcription regulation in mammalian cells, and it shows that the baculoviral genome can be activated in a mammalian system, and also that the two major -activators, IE1 and IE2, play a central role in this activation.

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2007-08-01
2024-03-28
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References

  1. Abe T., Takahashi H., Hamazaki H., Miyano-Kurosaki N., Matsuura Y., Takaku H. 2003; Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J Immunol 171:1133–1139 [CrossRef]
    [Google Scholar]
  2. Abe T., Hemmi H., Miyamoto H., Moriishi K., Tamura S., Takaku H., Akira S., Matsuura Y. 2005; Involvement of the Toll-like receptor 9 signaling pathway in the induction of innate immunity by baculovirus. J Virol 79:2847–2858 [CrossRef]
    [Google Scholar]
  3. 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 [CrossRef]
    [Google Scholar]
  4. Blissard G. W., Rohrmann G. F. 1991; Baculovirus gp64 gene expression: analysis of sequences modulating early transcription and transactivation by IE1. J Virol 65:5820–5827
    [Google Scholar]
  5. Boyce F. M., Bucher N. L. 1996; Baculovirus-mediated gene transfer into mammalian cells. Proc Natl Acad Sci U S A 93:2348–2352 [CrossRef]
    [Google Scholar]
  6. Carson D. D., Guarino L. A., Summers M. D. 1988; Functional mapping of an AcNPV immediately early gene which augments expression of the IE1 trans-activated 39K gene. Virology 162:444–451 [CrossRef]
    [Google Scholar]
  7. Carson D. D., Summers M. D., Guarino L. A. 1991; Molecular analysis of a baculovirus regulatory gene. Virology 182:279–286 [CrossRef]
    [Google Scholar]
  8. Chang Y. J., Liu C. Y., Chiang B. L., Chao Y. C., Chen C. C. 2004; Induction of IL-8 release in lung cells via activator protein-1 by recombinant baculovirus displaying severe acute respiratory syndrome-coronavirus spike proteins: identification of two functional regions. J Immunol 173:7602–7614 [CrossRef]
    [Google Scholar]
  9. Choi J., Guarino L. A. 1995; The baculovirus transactivator IE1 binds to viral enhancer elements in the absence of insect cell factors. J Virol 69:4548–4551
    [Google Scholar]
  10. Dai X., Willis L. G., Huijskens I., Palli S. R., Theilmann D. A. 2004; The acidic activation domains of the baculovirus transactivators IE1 and IE0 are functional for transcriptional activation in both insect and mammalian cells. J Gen Virol 85:573–582 [CrossRef]
    [Google Scholar]
  11. Fujita R., Matsuyama T., Yamagishi J., Sahara K., Asano S., Bando H. 2006; Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and upregulation of the host β-actin gene. J Virol 80:2390–2395 [CrossRef]
    [Google Scholar]
  12. Gossen M., Bujard H. 1992; Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A 89:5547–5551 [CrossRef]
    [Google Scholar]
  13. Guarino L. A., Summers M. D. 1986; Functional mapping of a trans -activating gene required for expression of a baculovirus delayed-early gene. J Virol 57:563–571
    [Google Scholar]
  14. Hills D., Crane-Robinson C. 1995; Baculovirus expression of human basic fibroblast growth factor from a synthetic gene: role of the Kozak consensus and comparison with bacterial expression. Biochim Biophys Acta 1260:14–20 [CrossRef]
    [Google Scholar]
  15. Hofmann C., Sandig V., Jennings G., Rudolph M., Schlag P., Strauss M. 1995; Efficient gene transfer into human hepatocytes by baculovirus vectors. Proc Natl Acad Sci U S A 92:10099–10103 [CrossRef]
    [Google Scholar]
  16. Hu Y. C., Tsai C. T., Chang Y. J., Huang J. H. 2003; Enhancement and prolongation of baculovirus-mediated expression in mammalian cells: focuses on strategic infection and feeding. Biotechnol Prog 19:373–379 [CrossRef]
    [Google Scholar]
  17. Huser A., Hofmann C. 2003; Baculovirus vectors: novel mammalian cell gene-delivery vehicles and their applications. Am J Pharmacogenomics 3:53–63
    [Google Scholar]
  18. Huser A., Rudolph M., Hofmann C. 2001; Incorporation of decay-accelerating factor into the baculovirus envelope generates complement-resistant gene transfer vectors. Nat Biotechnol 19:451–455 [CrossRef]
    [Google Scholar]
  19. Kenoutis C., Efrose R. C., Swevers L., Lavdas A. A., Gaitanou M., Matsas R., Iatrou K. 2006; Baculovirus-mediated gene delivery into mammalian cells does not alter their transcriptional and differentiating potential but is accompanied by early viral gene expression. J Virol 80:4135–4146 [CrossRef]
    [Google Scholar]
  20. Kost T. A., Condreay J. P., Jarvis D. L. 2005; Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 23:567–575 [CrossRef]
    [Google Scholar]
  21. Kovacs G. R., Choi J., Guarino L. A., Summers M. D. 1992; Functional dissection of the Autographa californic a nuclear polyhedrosis virus immediate-early 1 transcriptional regulatory protein. J Virol 66:7429–7437
    [Google Scholar]
  22. Kozak M. 1987; An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148 [CrossRef]
    [Google Scholar]
  23. Leisy D. J., Rasmussen C., Kim H. T., Rohrmann G. F. 1995; The Autographa californica nuclear polyhedrosis virus homologous region 1a: identical sequences are essential for DNA replication activity and transcriptional enhancer function. Virology 208:742–752 [CrossRef]
    [Google Scholar]
  24. Leisy D. J., Rasmussen C., Owusu E. O., Rohrmann G. F. 1997; A mechanism for negative gene regulation in Autographa californica multinucleocapsid nuclear polyhedrosis virus. J Virol 71:5088–5094
    [Google Scholar]
  25. Lo H. R., Chao Y. C. 2004; Rapid titer determination of baculovirus by quantitative real-time polymerase chain reaction. Biotechnol Prog 20:354–360
    [Google Scholar]
  26. Lo H. R., Chou C. C., Wu T. Y., Yuen J. P., Chao Y. C. 2002; Novel baculovirus DNA elements strongly stimulate activities of exogenous and endogenous promoters. J Biol Chem 277:5256–5264 [CrossRef]
    [Google Scholar]
  27. Lu A., Carstens E. B. 1993; Immediate-early baculovirus genes transactivate the p143 gene promoter of Autographa californica nuclear polyhedrosis virus. Virology 195:710–718 [CrossRef]
    [Google Scholar]
  28. Lu A., Miller L. K. 1995; The roles of eighteen baculovirus late expression factor genes in transcription and DNA replication. J Virol 69:975–982
    [Google Scholar]
  29. Murges D., Kremer A., Knebel-Morsdorf D. 1997; Baculovirus transactivator IE1 is functional in mammalian cells. J Gen Virol 78:1507–1510
    [Google Scholar]
  30. Nissen M. S., Friesen P. D. 1989; Molecular analysis of the transcriptional regulatory region of an early baculovirus gene. J Virol 63:493–503
    [Google Scholar]
  31. O'Reilly D. R., Miller L. K., Luckow V. A. 1994 Baculovirus Expression Vectors: a Laboratory Manual New York: Oxford University Press;
    [Google Scholar]
  32. Prikhod'ko E. A., Lu A., Wilson J. A., Miller L. K. 1999; In vivo and in vitro analysis of baculovirus ie-2 mutants. J Virol 73:2460–2468
    [Google Scholar]
  33. Shoji I., Aizaki H., Tani H., Ishii K., Chiba T., Saito I., Miyamura T., Matsuura Y. 1997; Efficient gene transfer into various mammalian cells, including non-hepatic cells, by baculovirus vectors. J Gen Virol 78:2657–2664
    [Google Scholar]
  34. Yoo S., Guarino L. A. 1994; Functional dissection of the ie2 gene product of the baculovirus Autographa californica nuclear polyhedrosis virus. Virology 202:164–172 [CrossRef]
    [Google Scholar]
  35. Zheng D., Constantinidou C., Hobman J. L., Minchin S. D. 2004; Identification of the CRP regulon using in vitro and in vivo transcriptional profiling. Nucleic Acids Res 32:5874–5893 [CrossRef]
    [Google Scholar]
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