Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1995 Mar 1;181(3):1217–1222. doi: 10.1084/jem.181.3.1217

Calcium/calmodulin-dependent protein kinase II downregulates both calcineurin and protein kinase C-mediated pathways for cytokine gene transcription in human T cells

PMCID: PMC2191942  PMID: 7869038

Abstract

Engagement of the T cell receptor for antigen activates phospholipase C resulting in an increase in intracellular free calcium concentration ([Ca2+]i) and activation of protein kinase C (PKC). Increased [Ca2+]i activates Ca2+/calmodulin-dependent kinases including the multifunctional Ca2+/calmodulin-dependent protein kinase II (CaM-K II), as well as calcineurin, a type 2B protein phosphatase. Recent studies have identified calcineurin as a key enzyme for interleukin (IL)-2 and IL-4 promoter activation. However, the role of CaM-K II remains unknown. We have used mutants of these kinases and phosphatases (gamma B*CaM-K and delta CaM-AI, respectively) to explore their relative role in cytokine gene transcription and their interactions with PKC- dependent signaling systems. gamma B*CaM-K and delta CaM-AI, known to exhibit constitutive Ca(2+)-independent activity, were cotransfected (alone or in combination) in Jurkat T cells with a plasmid containing the intact IL-2 promoter driving the expression of the chloramphenicol acetyltransferase reporter gene. Cotransfection of gamma B*CaM-K with the IL-2 promoter construct downregulated its transcription in response to stimulation with ionomycin and phorbol myristate acetate (PMA). The inhibitory effect of CaM-K II on IL-2 promoter was associated with decreased transcription of its AP-1 and NF-AT transactivating pathways. Under the same conditions, delta CaM-AI superinduced IL-2 promoter activity (approximately twofold increase). When both mutants were used in combination, gamma B*CaM-K inhibited the induction of the IL-2 promoter by delta CaM-AI. Similar results were obtained when a construct containing the IL-4 promoter also was used. gamma B*CaM-K also downregulated the activation of AP-1 in response to transfection with a constitutively active mutant of PKC or stimulation with PMA. These results suggest that CaM-K II may exert negative influences on cytokine gene transcription in human T cells, and provide preliminary evidence for negative cross-talk with the calcineurin- and PKC- dependent signaling systems.

Full Text

The Full Text of this article is available as a PDF (652.4 KB).

Selected References

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

  1. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  2. Clipstone N. A., Crabtree G. R. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature. 1992 Jun 25;357(6380):695–697. doi: 10.1038/357695a0. [DOI] [PubMed] [Google Scholar]
  3. Edman C. F., Schulman H. Identification and characterization of delta B-CaM kinase and delta C-CaM kinase from rat heart, two new multifunctional Ca2+/calmodulin-dependent protein kinase isoforms. Biochim Biophys Acta. 1994 Mar 10;1221(1):89–101. doi: 10.1016/0167-4889(94)90221-6. [DOI] [PubMed] [Google Scholar]
  4. Frantz B., Nordby E. C., Bren G., Steffan N., Paya C. V., Kincaid R. L., Tocci M. J., O'Keefe S. J., O'Neill E. A. Calcineurin acts in synergy with PMA to inactivate I kappa B/MAD3, an inhibitor of NF-kappa B. EMBO J. 1994 Feb 15;13(4):861–870. doi: 10.1002/j.1460-2075.1994.tb06329.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fraser J. D., Irving B. A., Crabtree G. R., Weiss A. Regulation of interleukin-2 gene enhancer activity by the T cell accessory molecule CD28. Science. 1991 Jan 18;251(4991):313–316. doi: 10.1126/science.1846244. [DOI] [PubMed] [Google Scholar]
  6. Go C., Miller J. Differential induction of transcription factors that regulate the interleukin 2 gene during anergy induction and restimulation. J Exp Med. 1992 May 1;175(5):1327–1336. doi: 10.1084/jem.175.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hanson P. I., Schulman H. Neuronal Ca2+/calmodulin-dependent protein kinases. Annu Rev Biochem. 1992;61:559–601. doi: 10.1146/annurev.bi.61.070192.003015. [DOI] [PubMed] [Google Scholar]
  8. Harding F. A., McArthur J. G., Gross J. A., Raulet D. H., Allison J. P. CD28-mediated signalling co-stimulates murine T cells and prevents induction of anergy in T-cell clones. Nature. 1992 Apr 16;356(6370):607–609. doi: 10.1038/356607a0. [DOI] [PubMed] [Google Scholar]
  9. Hashimoto Y., Soderling T. R. Regulation of calcineurin by phosphorylation. Identification of the regulatory site phosphorylated by Ca2+/calmodulin-dependent protein kinase II and protein kinase C. J Biol Chem. 1989 Oct 5;264(28):16524–16529. [PubMed] [Google Scholar]
  10. Kang S. M., Beverly B., Tran A. C., Brorson K., Schwartz R. H., Lenardo M. J. Transactivation by AP-1 is a molecular target of T cell clonal anergy. Science. 1992 Aug 21;257(5073):1134–1138. doi: 10.1126/science.257.5073.1134. [DOI] [PubMed] [Google Scholar]
  11. Kapiloff M. S., Mathis J. M., Nelson C. A., Lin C. R., Rosenfeld M. G. Calcium/calmodulin-dependent protein kinase mediates a pathway for transcriptional regulation. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3710–3714. doi: 10.1073/pnas.88.9.3710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kubo M., Kincaid R. L., Webb D. R., Ransom J. T. The Ca2+/calmodulin-activated, phosphoprotein phosphatase calcineurin is sufficient for positive transcriptional regulation of the mouse IL-4 gene. Int Immunol. 1994 Feb;6(2):179–188. doi: 10.1093/intimm/6.2.179. [DOI] [PubMed] [Google Scholar]
  13. MacNicol M., Schulman H. Cross-talk between protein kinase C and multifunctional Ca2+/calmodulin-dependent protein kinase. J Biol Chem. 1992 Jun 15;267(17):12197–12201. [PubMed] [Google Scholar]
  14. Muramatsu M., Kaibuchi K., Arai K. A protein kinase C cDNA without the regulatory domain is active after transfection in vivo in the absence of phorbol ester. Mol Cell Biol. 1989 Feb;9(2):831–836. doi: 10.1128/mcb.9.2.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakayama T., Ueda Y., Yamada H., Shores E. W., Singer A., June C. H. In vivo calcium elevations in thymocytes with T cell receptors that are specific for self ligands. Science. 1992 Jul 3;257(5066):96–99. doi: 10.1126/science.1621102. [DOI] [PubMed] [Google Scholar]
  16. Nghiem P., Ollick T., Gardner P., Schulman H. Interleukin-2 transcriptional block by multifunctional Ca2+/calmodulin kinase. Nature. 1994 Sep 22;371(6495):347–350. doi: 10.1038/371347a0. [DOI] [PubMed] [Google Scholar]
  17. Nghiem P., Saati S. M., Martens C. L., Gardner P., Schulman H. Cloning and analysis of two new isoforms of multifunctional Ca2+/calmodulin-dependent protein kinase. Expression in multiple human tissues. J Biol Chem. 1993 Mar 15;268(8):5471–5479. [PubMed] [Google Scholar]
  18. O'Keefe S. J., Tamura J., Kincaid R. L., Tocci M. J., O'Neill E. A. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin. Nature. 1992 Jun 25;357(6380):692–694. doi: 10.1038/357692a0. [DOI] [PubMed] [Google Scholar]
  19. Paliogianni F., Kincaid R. L., Boumpas D. T. Prostaglandin E2 and other cyclic AMP elevating agents inhibit interleukin 2 gene transcription by counteracting calcineurin-dependent pathways. J Exp Med. 1993 Nov 1;178(5):1813–1817. doi: 10.1084/jem.178.5.1813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Paliogianni F., Raptis A., Ahuja S. S., Najjar S. M., Boumpas D. T. Negative transcriptional regulation of human interleukin 2 (IL-2) gene by glucocorticoids through interference with nuclear transcription factors AP-1 and NF-AT. J Clin Invest. 1993 Apr;91(4):1481–1489. doi: 10.1172/JCI116353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schwartz R. H. Costimulation of T lymphocytes: the role of CD28, CTLA-4, and B7/BB1 in interleukin-2 production and immunotherapy. Cell. 1992 Dec 24;71(7):1065–1068. doi: 10.1016/s0092-8674(05)80055-8. [DOI] [PubMed] [Google Scholar]
  22. Tamura T., Yanagida T., Nariuchi H. Difference in signal transduction pathway for IL-2 and IL-4 production in T helper 1 and T helper 2 cell clones in response to anti-CD3. J Immunol. 1993 Dec 1;151(11):6051–6061. [PubMed] [Google Scholar]
  23. Ullman K. S., Northrop J. P., Admon A., Crabtree G. R. Jun family members are controlled by a calcium-regulated, cyclosporin A-sensitive signaling pathway in activated T lymphocytes. Genes Dev. 1993 Feb;7(2):188–196. doi: 10.1101/gad.7.2.188. [DOI] [PubMed] [Google Scholar]
  24. Ullman K. S., Northrop J. P., Verweij C. L., Crabtree G. R. Transmission of signals from the T lymphocyte antigen receptor to the genes responsible for cell proliferation and immune function: the missing link. Annu Rev Immunol. 1990;8:421–452. doi: 10.1146/annurev.iy.08.040190.002225. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES