Attenuated Expression of Interferon-induced Protein Kinase PKR in a Simian Cell Devoid of Type I Interferons

  • Park, Se-Hoon (School of Life Sciences and Biotechnology, Korea University) ;
  • Choi, Jaydo (School of Life Sciences and Biotechnology, Korea University) ;
  • Kang, Ju-Il (School of Life Sciences and Biotechnology, Korea University) ;
  • Choi, Sang-Yun (School of Life Sciences and Biotechnology, Korea University) ;
  • Hwang, Soon-Bong (Ilsong Institute of Life Science, Hallym University) ;
  • Kim, Jungsuh P. (School of Life Sciences and Biotechnology, Korea University) ;
  • Ahn, Byung-Yoon (Ilsong Institute of Life Science, Hallym University)
  • 투고 : 2005.07.11
  • 심사 : 2005.10.13
  • 발행 : 2006.02.28

초록

The interferon-induced, double-stranded RNA (dsRNA)-dependent protein kinase PKR plays a key role in interferon-mediated host defense against viral infection, and is implicated in cellular transformation and apoptosis. We have isolated a cDNA of simian PKR encoding a product with 83% amino acid identity to the human homolog and showed that PKR expression is significantly attenuated in the Vero E6 African green monkey kidney cells devoid of type I interferon genes. A variant form of PKR lacking the exon 12 in the kinase domain is produced in these cells, presumably from an alternatively spliced transcript. Unlike wild type PKR, the variant protein named PKR-${\Delta}E12$ is incapable of auto-phosphorylation and phosphorylation of eIF2-${\alpha}$, indicating that the kinase sub-domains III and IV embedded in exon 12 are indispensable for catalytic function. PKR-${\Delta}E12$ had no dominant negative effect but was weakly phosphorylated in trans by wild type PKR.

키워드

과제정보

연구 과제 주관 기관 : Korea Science and Engineering Foundation, Ministry of Health and Welfare

참고문헌

  1. Abraham, N., Jaramillo, M. L., Duncan, P. I., Methot, N., Icely, P. L., et al. (1998) The murine PKR tumor suppressor gene is rearranged in a lymphocytic leukemia. Exp. Cell Res. 244, 394−404 https://doi.org/10.1006/excr.1998.4201
  2. Baltzis, D., Li, S., and Koromilas, A. E. (2002) Functional characterization of pkr gene products expressed in cells from mice with a targeted deletion of the N terminus or C terminus domain of PKR. J. Biol. Chem. 277, 38364–38372 https://doi.org/10.1074/jbc.M203564200
  3. Barber, G. N., Wambach, M., Thompson, S., Jagus, R., and Katze, M. G. (1995) Mutants of the RNA-dependent protein kinase (PKR) lacking double-stranded RNA binding domain I can act as transdominant inhibitors and induce malignant transformation. Mol. Cell. Biol. 15, 3138-3146
  4. Clemens, M. J. and Androulla, E. (1997) The double-stranded RNA-dependent protein kinase PKR: structure and function. J. Interferon Cytokine Res. 17, 503-524 https://doi.org/10.1089/jir.1997.17.503
  5. Diaz, M. O., Ziemin, S., Le Beau, M. M., Pitha, P., Smith, S. D., et al. (1988) Homologous deletion of the alpha- and beta 1 interferon genes in human leukemia and derived cell lines. Proc. Natl. Acad. Sci. USA 85, 5259–5263
  6. Feng, G. S., Chong, K., Kumar, A., and Williams, B. R. G. (1992) Identification of double-stranded RNA-binding domains in the interferon-induced double-stranded RNAactivated p68 kinase. Proc. Natl. Acad. Sci. USA 89, 5447– 5451
  7. Frohman, M. A., Dush, M. K., and Martin, G. R. (1988) Rapid production of full-length cDNA from rare transcripts: Amplification using a single gene specific oligonucleotide primer. Proc. Natl. Acad. Sci. USA 85, 8998–9002
  8. Gale, M. Jr. and Katze, M. J. (1998) Molecular mechanisms of interferon resistance mediated by viral-directed inhibition of PKR, the interferon-induced protein kinase. Pharmacol. Ther. 78, 29-46 https://doi.org/10.1016/S0163-7258(97)00165-4
  9. Hershey, J. W. B. (1989) Protein phosphorylation controls translation rates. J. Biol. Chem. 264, 20823–20826
  10. Icely, P. L., Gross, P., Bergeron, J. J. M., Devault, A., Afar, D. E. H., et al. (1991) TIK, a novel serine/threonine kinase, is recognized by antibodies directed against phosphotyrosine. J. Biol. Chem. 266, 16073–16077
  11. Katze, M. G., Wambach, M., Wong, M.-L., Garfinkel, M., Meurs, E., et al. (1991) Functional expression and RNA binding analysis of the interferon-induced, double-stranded RNA-activated, 68,000-Mr protein kinase in a cell free system. Mol. Cell. Biol. 11, 5497-5505
  12. Koromilas, A. E., Roy, S., Barber, G. N., Katze, M. G., and Sonenberg, N. (1992) Malignant transformation by a mutant of the IFN-inducible dsRNA-dependent protein kinase. Science 257, 1685-1689 https://doi.org/10.1126/science.1382315
  13. Kumar, A., Haque, J., Lacoste, J., Hiscott, J., and Williams, B. R. (1994) Double-stranded RNA-dependent protein kinase activates transcription factor NF-kappa B by phosphorylating I kappa B. Proc. Natl. Acad. Sci. USA 91, 6288–6292
  14. Lee, S. B. and Esteban, M. (1993) The interferon-induced double- stranded RNA-activated human p68 protein kinase inhibits the replication of vaccinia virus. Virology 193, 1037-1041 https://doi.org/10.1006/viro.1993.1223
  15. Li, S. and Koromilas, A. E. (2001) Dominant negative function by an alternative spliced form of the interferon-inducible protein kinase PKR. J. Biol. Chem. 276, 13881-13890
  16. Mellor, H., Flowers, K. M., Kimball, S. R., and Jefferson, L. S. (1994) Cloning and characterization of a cDNA encoding rat PKR, the double-stranded RNA-dependent eukaryotic initiation factor-2 kinase. Biochim. Biophys. Acta 1219, 693-696 https://doi.org/10.1016/0167-4781(94)90229-1
  17. Meurs, E., Chang, K., Galabru, J., Thomas, N. S., Kerr, I. M., et al. (1990) Molecular cloning and characterization of the human double-stranded RNA-activated protein kinase induced by interferon. Cell 62, 379-390 https://doi.org/10.1016/0092-8674(90)90374-N
  18. Meurs, E. F., Wantanabe, Y., Kadereit, S., Barber, G. N., Katze, M. G., et al. (1992) Constitutive expression of human double- stranded RNA-activated p68 kinase in murine cells mediates phosphorylation of eukaryotic initiation factor 2 and partial resistance to encephalomyocarditis virus growth. J. Virol. 66, 5805-5814
  19. Meurs, E. F., Galabru, J., Barber, G. N., Katze, M. G., and Hovanessian, A. G. (1993) Tumor suppressor function of the interferon-induced double-stranded RNA-activated protein kinase. Proc. Natl. Acad. Sci. USA 90, 232–236
  20. Samuel, C. E. (1979) Mechanism of interferon action: phosphorylation of protein synthesis initiation factor eIF-2 in interferon- treated human cells by a ribosome-associated protein kinase possessing site-specificity similar to heminregulated rabbit reticulocyte kinase. Proc. Natl. Acad. Sci. USA 76, 600–604
  21. Samuel, C. E. (1993) The eIF-2${\alpha}$ protein kinases, regulators of translation in eukaryotes from yeasts to humans. J. Biol. Chem. 268, 7603-7606
  22. Tan, S. L. and Katze, M. G. (1999) The emerging role of the interferon-induced PKR protein kinase as an apoptotic effector: a new face of death- J. Interferon Cytokine Res. 19, 543-554 https://doi.org/10.1089/107999099313677
  23. Thomis, D. C., Doohan, J. D., and Samuel, C. E. (1992) Mechanism of interferon action: cDNA structure, expression and regulation of the interferon-induced, RNA-dependent PI/eIF-2${\alpha}$ protein kinase from human cells. Virology 188, 33-46 https://doi.org/10.1016/0042-6822(92)90732-5
  24. Xu, Z. and Williams, B. R. (1998) Genomic features of human PKR: alternative splicing and a polymorphic CGG repeat in the 5′-untranslated region. J. Interferon Cytokine Res. 18, 609-616 https://doi.org/10.1089/jir.1998.18.609