Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Nov;92(5):2207–2214. doi: 10.1172/JCI116823

Early events of human T lymphocyte activation are associated with type I protein kinase A activity.

D Laxminarayana 1, A Berrada 1, G M Kammer 1
PMCID: PMC288400  PMID: 8227335

Abstract

Human T lymphocytes possess both the type I and II isozymes of protein kinase A (PKA). The type I (PKA-I) isozyme is predominantly associated with the plasma membrane, whereas the type II (PKA-II) isozyme is primarily localized to the cytosol. Because the functions of both PKA-I and PKA-II isozymes in the biochemical events of T lymphocyte activation have not been clearly elucidated, we tested the hypothesis that very early events of normal human T lymphocyte activation are mediated by the PKA-I and/or PKA-II isozyme(s). Fresh normal human T cells and a normal human CD4+ T cell line (GK606) activated with anti-CD3-epsilon and recombinant interleukin 1 alpha (rIL-1 alpha) exhibited a peak six- to sevenfold increase of PKA phosphotransferase activity at 5 min that returned to baseline by 60 min. Similarly, both fresh T cells and the T cell line activated by phorbol myristate acetate and ionomycin demonstrated a peak eightfold increase of PKA activity by 15 min that returned toward baseline by 60 min. Chromatographic separation of the PKA isozymes and quantification of phosphotransferase activities after T cell activation by either agonist pair showed preferential activation of the PKA-I isozyme, resulting in a significant reduction in the ratio of PKA-I to PKA-II isozyme activity from 3.1:1-6.2:1 to 1.1:1-3.2:1. PKA-I isozyme activation resulted in the release of free catalytic (C) subunit, an increase in C subunit phosphotransferase activity, and the phosphorylation of T cell plasma membrane-associated proteins, p14, p17, p20, p21, p38, and p48. However, activation of the PKA-I isozyme did not appear to be required for the transcription of IL-2 mRNA, an event necessary for mitosis. These data indicate that ligand-induced T cell activation is associated with rapid activation of the PKA-I, but not PKA-II, isozyme that results in phosphorylation of plasma membrane-associated proteins. The involvement of the PKA-I isozyme during the very early events of T cell activation suggests that this isozyme may be an antigen- or mitogen-stimulated protein kinase.

Full text

PDF
2207

Images in this article

2209Image
on p.2209
2209Image
on p.2209
2212Image
on p.2212

Selected References

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

  1. Bell J. D., Brunton L. L. Enhancement of adenylate cyclase activity in S49 lymphoma cells by phorbol esters. Withdrawal of GTP-dependent inhibition. J Biol Chem. 1986 Sep 15;261(26):12036–12041. [PubMed] [Google Scholar]
  2. Berry N., Nishizuka Y. Protein kinase C and T cell activation. Eur J Biochem. 1990 Apr 30;189(2):205–214. doi: 10.1111/j.1432-1033.1990.tb15478.x. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Braylan R. C., Benson N. A., Nourse V., Kruth H. S. Correlated analysis of cellular DNA, membrane antigens and light scatter of human lymphoid cells. Cytometry. 1982 Mar;2(5):337–343. doi: 10.1002/cyto.990020511. [DOI] [PubMed] [Google Scholar]
  5. Brenner C. A., Tam A. W., Nelson P. A., Engleman E. G., Suzuki N., Fry K. E., Larrick J. W. Message amplification phenotyping (MAPPing): a technique to simultaneously measure multiple mRNAs from small numbers of cells. Biotechniques. 1989 Nov-Dec;7(10):1096–1103. [PubMed] [Google Scholar]
  6. Byus C. V., Klimpel G. R., Lucas D. O., Russell D. H. Type I and type II cyclic AMP-dependent protein kinase as opposite effectors of lymphocyte mitogenesis. Nature. 1977 Jul 7;268(5615):63–64. doi: 10.1038/268063a0. [DOI] [PubMed] [Google Scholar]
  7. Cantrell D. A., Davies A. A., Crumpton M. J. Activators of protein kinase C down-regulate and phosphorylate the T3/T-cell antigen receptor complex of human T lymphocytes. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8158–8162. doi: 10.1073/pnas.82.23.8158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cantrell D., Davies A. A., Londei M., Feldman M., Crumpton M. J. Association of phosphorylation of the T3 antigen with immune activation of T lymphocytes. Nature. 1987 Feb 5;325(6104):540–542. doi: 10.1038/325540a0. [DOI] [PubMed] [Google Scholar]
  9. Chedid M., Mizel S. B. Involvement of cyclic AMP-dependent protein kinases in the signal transduction pathway for interleukin-1. Mol Cell Biol. 1990 Jul;10(7):3824–3827. doi: 10.1128/mcb.10.7.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chedid M., Yoza B. K., Brooks J. W., Mizel S. B. Activation of AP-1 by IL-1 and phorbol esters in T cells. Role of protein kinase A and protein phosphatases. J Immunol. 1991 Aug 1;147(3):867–873. [PubMed] [Google Scholar]
  11. Crabtree G. R. Contingent genetic regulatory events in T lymphocyte activation. Science. 1989 Jan 20;243(4889):355–361. doi: 10.1126/science.2783497. [DOI] [PubMed] [Google Scholar]
  12. Didier M., Aussel C., Pelassy C., Fehlmann M. IL-1 signaling for IL-2 production in T cells involves a rise in phosphatidylserine synthesis. J Immunol. 1988 Nov 1;141(9):3078–3080. [PubMed] [Google Scholar]
  13. Grove J. R., Deutsch P. J., Price D. J., Habener J. F., Avruch J. Plasmids encoding PKI(1-31), a specific inhibitor of cAMP-stimulated gene expression, inhibit the basal transcriptional activity of some but not all cAMP-regulated DNA response elements in JEG-3 cells. J Biol Chem. 1989 Nov 25;264(33):19506–19513. [PubMed] [Google Scholar]
  14. Hasler P., Moore J. J., Kammer G. M. Human T lymphocyte cAMP-dependent protein kinase: subcellular distributions and activity ranges of type I and type II isozymes. FASEB J. 1992 Jun;6(9):2735–2741. doi: 10.1096/fasebj.6.9.1319361. [DOI] [PubMed] [Google Scholar]
  15. Hasler P., Schultz L. A., Kammer G. M. Defective cAMP-dependent phosphorylation of intact T lymphocytes in active systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1978–1982. doi: 10.1073/pnas.87.5.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hidaka H., Kobayashi R. Pharmacology of protein kinase inhibitors. Annu Rev Pharmacol Toxicol. 1992;32:377–397. doi: 10.1146/annurev.pa.32.040192.002113. [DOI] [PubMed] [Google Scholar]
  17. Hollingsworth E. B., Ukena D., Daly J. W. The protein kinase C activator phorbol-12-myristate-13-acetate enhances cyclic AMP accumulation in pheochromocytoma cells. FEBS Lett. 1986 Feb 3;196(1):131–134. doi: 10.1016/0014-5793(86)80227-7. [DOI] [PubMed] [Google Scholar]
  18. Kammer G. M., Boehm C. A., Rudolph S. A., Schultz L. A. Mobility of the human T lymphocyte surface molecules CD3, CD4, and CD8: regulation by a cAMP-dependent pathway. Proc Natl Acad Sci U S A. 1988 Feb;85(3):792–796. doi: 10.1073/pnas.85.3.792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kammer G. M., Kurrasch R., Scillian J. J. Capping of the surface OKT3 binding molecule prevents the T-cell proliferative response to antigens: evidence that this molecule conveys the activation signal. Cell Immunol. 1984 Aug;87(1):284–294. doi: 10.1016/0008-8749(84)90152-7. [DOI] [PubMed] [Google Scholar]
  20. Kammer G. M. The adenylate cyclase-cAMP-protein kinase A pathway and regulation of the immune response. Immunol Today. 1988 Jul-Aug;9(7-8):222–229. doi: 10.1016/0167-5699(88)91220-0. [DOI] [PubMed] [Google Scholar]
  21. Katada T., Gilman A. G., Watanabe Y., Bauer S., Jakobs K. H. Protein kinase C phosphorylates the inhibitory guanine-nucleotide-binding regulatory component and apparently suppresses its function in hormonal inhibition of adenylate cyclase. Eur J Biochem. 1985 Sep 2;151(2):431–437. doi: 10.1111/j.1432-1033.1985.tb09120.x. [DOI] [PubMed] [Google Scholar]
  22. Klausner R. D., O'Shea J. J., Luong H., Ross P., Bluestone J. A., Samelson L. E. T cell receptor tyrosine phosphorylation. Variable coupling for different activating ligands. J Biol Chem. 1987 Sep 15;262(26):12654–12659. [PubMed] [Google Scholar]
  23. Klausner R. D., Samelson L. E. T cell antigen receptor activation pathways: the tyrosine kinase connection. Cell. 1991 Mar 8;64(5):875–878. doi: 10.1016/0092-8674(91)90310-u. [DOI] [PubMed] [Google Scholar]
  24. Livesey S. A., Martin T. J. Selective activation of the cAMP-dependent protein kinase isoenzymes. Methods Enzymol. 1988;159:105–118. doi: 10.1016/0076-6879(88)59012-2. [DOI] [PubMed] [Google Scholar]
  25. Lohmann S. M., Walter U. Regulation of the cellular and subcellular concentrations and distribution of cyclic nucleotide-dependent protein kinases. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;18:63–117. [PubMed] [Google Scholar]
  26. Muñoz E., Zubiaga A., Huang C., Huber B. T. Interleukin-1 induces protein tyrosine phosphorylation in T cells. Eur J Immunol. 1992 Jun;22(6):1391–1396. doi: 10.1002/eji.1830220610. [DOI] [PubMed] [Google Scholar]
  27. Naghshineh S., Noguchi M., Huang K. P., Londos C. Activation of adipocyte adenylate cyclase by protein kinase C. J Biol Chem. 1986 Nov 5;261(31):14534–14538. [PubMed] [Google Scholar]
  28. Nikula H., Vihko K., Huhtaniemi I. Protein kinase C and Gi-protein mediated modulation of cAMP production in different stages of the rat seminiferous epithelium. Mol Cell Endocrinol. 1990 May 7;70(3):247–253. doi: 10.1016/0303-7207(90)90215-t. [DOI] [PubMed] [Google Scholar]
  29. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  30. O'Neill L. A., Bird T. A., Saklatvala J. How does interleukin 1 activate cells? Interleukin 1 signal transduction. Immunol Today. 1990 Nov;11(11):392–394. doi: 10.1016/0167-5699(90)90155-3. [DOI] [PubMed] [Google Scholar]
  31. Osborn L., Kunkel S., Nabel G. J. Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2336–2340. doi: 10.1073/pnas.86.7.2336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Palacios R. Mechanisms by which accessory cells contribute in growth of resting T lymphocytes initiated by OKT3 antibody. Eur J Immunol. 1985 Jul;15(7):645–651. doi: 10.1002/eji.1830150702. [DOI] [PubMed] [Google Scholar]
  33. Pyne N. J., Freissmuth M., Palmer S. Phosphorylation of the spliced variant forms of the recombinant stimulatory guanine-nucleotide-binding regulatory protein (Gs alpha) by protein kinase C. Biochem J. 1992 Jul 1;285(Pt 1):333–338. doi: 10.1042/bj2850333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ray K., Thompson N., Kennard N., Rollins P., Grenfell S., Witham S., Smithers N., Solari R. Investigation of guanine-nucleotide-binding protein involvement and regulation of cyclic AMP metabolism in interleukin 1 signal transduction. Biochem J. 1992 Feb 15;282(Pt 1):59–67. doi: 10.1042/bj2820059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rollins P., Witham S., Ray K., Thompson N., Sadler H., Smithers N., Grenfell S., Solari R. Modification of biological responses to interleukin-1 by agents that perturb signal transduction pathways. Cytokine. 1991 Jan;3(1):42–53. doi: 10.1016/1043-4666(91)90009-3. [DOI] [PubMed] [Google Scholar]
  36. Rothermel J. D., Jastorff B., Botelho L. H. Inhibition of glucagon-induced glycogenolysis in isolated rat hepatocytes by the Rp diastereomer of adenosine cyclic 3',5'-phosphorothioate. J Biol Chem. 1984 Jul 10;259(13):8151–8155. [PubMed] [Google Scholar]
  37. Samelson L. E., Patel M. D., Weissman A. M., Harford J. B., Klausner R. D. Antigen activation of murine T cells induces tyrosine phosphorylation of a polypeptide associated with the T cell antigen receptor. Cell. 1986 Sep 26;46(7):1083–1090. doi: 10.1016/0092-8674(86)90708-7. [DOI] [PubMed] [Google Scholar]
  38. Shirakawa F., Chedid M., Suttles J., Pollok B. A., Mizel S. B. Interleukin 1 and cyclic AMP induce kappa immunoglobulin light-chain expression via activation of an NF-kappa B-like DNA-binding protein. Mol Cell Biol. 1989 Mar;9(3):959–964. doi: 10.1128/mcb.9.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shirakawa F., Yamashita U., Chedid M., Mizel S. B. Cyclic AMP--an intracellular second messenger for interleukin 1. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8201–8205. doi: 10.1073/pnas.85.21.8201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tortora G., Yokozaki H., Pepe S., Clair T., Cho-Chung Y. S. Differentiation of HL-60 leukemia by type I regulatory subunit antisense oligodeoxynucleotide of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):2011–2015. doi: 10.1073/pnas.88.5.2011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Unanue E. R., Allen P. M. The basis for the immunoregulatory role of macrophages and other accessory cells. Science. 1987 May 1;236(4801):551–557. doi: 10.1126/science.2437650. [DOI] [PubMed] [Google Scholar]
  42. Weiss A., Imboden J. B. Cell surface molecules and early events involved in human T lymphocyte activation. Adv Immunol. 1987;41:1–38. doi: 10.1016/s0065-2776(08)60029-2. [DOI] [PubMed] [Google Scholar]
  43. Yang S. Y., Chouaib S., Dupont B. A common pathway for T lymphocyte activation involving both the CD3-Ti complex and CD2 sheep erythrocyte receptor determinants. J Immunol. 1986 Aug 15;137(4):1097–1100. [PubMed] [Google Scholar]
  44. Yatomi Y., Arata Y., Tada S., Kume S., Ui M. Phosphorylation of the inhibitory guanine-nucleotide-binding protein as a possible mechanism of inhibition by protein kinase C of agonist-induced Ca2+ mobilization in human platelet. Eur J Biochem. 1992 May 1;205(3):1003–1009. doi: 10.1111/j.1432-1033.1992.tb16867.x. [DOI] [PubMed] [Google Scholar]
  45. Yoshimasa T., Sibley D. R., Bouvier M., Lefkowitz R. J., Caron M. G. Cross-talk between cellular signalling pathways suggested by phorbol-ester-induced adenylate cyclase phosphorylation. Nature. 1987 May 7;327(6117):67–70. doi: 10.1038/327067a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES