Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Feb 15;88(4):1227–1231. doi: 10.1073/pnas.88.4.1227

Interleukin 2 induces tyrosine phosphorylation and activation of p72-74 Raf-1 kinase in a T-cell line.

B Turner 1, U Rapp 1, H App 1, M Greene 1, K Dobashi 1, J Reed 1
PMCID: PMC50990  PMID: 1996324

Abstract

Interleukin 2 (IL-2) is a lymphokine, produced by T cells upon antigenic or mitogenic stimulation, that is a critical regulator of T-cell proliferation. Although the binding of IL-2 to its receptor has been well characterized, the molecular mechanisms by which IL-2 transmits its signal from the membrane to the interior of the cell are poorly understood. Like most other growth factors, IL-2 causes rapid phosphorylation of proteins within its target cells. Unlike many other growth factors, however, the known subunits of the IL-2 receptor lack tyrosine-specific kinase activity, and little is known about the kinases whose activities are regulated by IL-2. Here we show that IL-2 (but not IL-4) induces rapid phosphorylation of the p72-74 serine/threonine-specific kinase encoded by the c-Raf-1 protooncogene in an IL-2-dependent murine T-cell line, CTLL-2, and that this phosphorylation is associated with increased kinase activity in p72-74 Raf-1-containing immune complexes. The concentration dependence of IL-2-mediated elevations in Raf-1 kinase activity correlated well with IL-2-stimulated proliferation of CTLL-2 cells. Furthermore, much of the IL-2-stimulated phosphorylation of p72-74 Raf-1 occurred on tyrosines. To our knowledge, the Raf-1 kinase represents the first endogenous substrate of an IL-2-regulated tyrosine kinase to be identified.

Full text

PDF
1227

Images in this article

1228Image
on p.1228
1229Image
on p.1229
1230Image
on p.1230
1230Image
on p.1230
1231Image
on p.1231

Selected References

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

  1. Carroll M. P., Clark-Lewis I., Rapp U. R., May W. S. Interleukin-3 and granulocyte-macrophage colony-stimulating factor mediate rapid phosphorylation and activation of cytosolic c-raf. J Biol Chem. 1990 Nov 15;265(32):19812–19817. [PubMed] [Google Scholar]
  2. Cooper J. A., Sefton B. M., Hunter T. Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 1983;99:387–402. doi: 10.1016/0076-6879(83)99075-4. [DOI] [PubMed] [Google Scholar]
  3. Daniel T. O., Tremble P. M., Frackelton A. R., Jr, Williams L. T. Purification of the platelet-derived growth factor receptor by using an anti-phosphotyrosine antibody. Proc Natl Acad Sci U S A. 1985 May;82(9):2684–2687. doi: 10.1073/pnas.82.9.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Farrar W. L., Ferris D. K. Two-dimensional analysis of interleukin 2-regulated tyrosine kinase activation mediated by the p70-75 beta subunit of the interleukin 2 receptor. J Biol Chem. 1989 Jul 25;264(21):12562–12567. [PubMed] [Google Scholar]
  5. Frackelton A. R., Jr, Ross A. H., Eisen H. N. Characterization and use of monoclonal antibodies for isolation of phosphotyrosyl proteins from retrovirus-transformed cells and growth factor-stimulated cells. Mol Cell Biol. 1983 Aug;3(8):1343–1352. doi: 10.1128/mcb.3.8.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gaulton G. N., Eardley D. D. Interleukin 2-dependent phosphorylation of interleukin 2 receptors and other T cell membrane proteins. J Immunol. 1986 Apr 1;136(7):2470–2477. [PubMed] [Google Scholar]
  7. Gelfand E. W., Mills G. B., Cheung R. K., Lee J. W., Grinstein S. Transmembrane ion fluxes during activation of human T lymphocytes: role of Ca2+, Na+/H+ exchange and phospholipid turnover. Immunol Rev. 1987 Feb;95:59–87. doi: 10.1111/j.1600-065x.1987.tb00500.x. [DOI] [PubMed] [Google Scholar]
  8. Gillis S., Smith K. A. Long term culture of tumour-specific cytotoxic T cells. Nature. 1977 Jul 14;268(5616):154–156. doi: 10.1038/268154a0. [DOI] [PubMed] [Google Scholar]
  9. Goodwin R. G., Friend D., Ziegler S. F., Jerzy R., Falk B. A., Gimpel S., Cosman D., Dower S. K., March C. J., Namen A. E. Cloning of the human and murine interleukin-7 receptors: demonstration of a soluble form and homology to a new receptor superfamily. Cell. 1990 Mar 23;60(6):941–951. doi: 10.1016/0092-8674(90)90342-c. [DOI] [PubMed] [Google Scholar]
  10. Greene W. C., Robb R. J., Svetlik P. B., Rusk C. M., Depper J. M., Leonard W. J. Stable expression of cDNA encoding the human interleukin 2 receptor in eukaryotic cells. J Exp Med. 1985 Jul 1;162(1):363–368. doi: 10.1084/jem.162.1.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hatakeyama M., Mori H., Doi T., Taniguchi T. A restricted cytoplasmic region of IL-2 receptor beta chain is essential for growth signal transduction but not for ligand binding and internalization. Cell. 1989 Dec 1;59(5):837–845. doi: 10.1016/0092-8674(89)90607-7. [DOI] [PubMed] [Google Scholar]
  12. Hatakeyama M., Tsudo M., Minamoto S., Kono T., Doi T., Miyata T., Miyasaka M., Taniguchi T. Interleukin-2 receptor beta chain gene: generation of three receptor forms by cloned human alpha and beta chain cDNA's. Science. 1989 May 5;244(4904):551–556. doi: 10.1126/science.2785715. [DOI] [PubMed] [Google Scholar]
  13. Ishii T., Takeshita T., Numata N., Sugamura K. Protein phosphorylation mediated by IL-2/IL-2 receptor beta-chain interaction. J Immunol. 1988 Jul 1;141(1):174–179. [PubMed] [Google Scholar]
  14. Itoh N., Yonehara S., Schreurs J., Gorman D. M., Maruyama K., Ishii A., Yahara I., Arai K., Miyajima A. Cloning of an interleukin-3 receptor gene: a member of a distinct receptor gene family. Science. 1990 Jan 19;247(4940):324–327. doi: 10.1126/science.2404337. [DOI] [PubMed] [Google Scholar]
  15. Kolch W., Weissinger E., Mischak H., Troppmair J., Showalter S. D., Lloyd P., Heidecker G., Rapp U. R. Probing structure and function of the raf protein kinase domain with monoclonal antibodies. Oncogene. 1990 May;5(5):713–720. [PubMed] [Google Scholar]
  16. Kondo S., Kinoshita M., Shimizu A., Saito Y., Konishi M., Sabe H., Honjo T. Expression and functional characterization of artificial mutants of interleukin-2 receptor. Nature. 1987 May 7;327(6117):64–67. doi: 10.1038/327064a0. [DOI] [PubMed] [Google Scholar]
  17. LeGrue S. J. Does interleukin 2 stimulus-response coupling result in generation of intracellular second messengers? Lymphokine Res. 1988 Summer;7(2):187–200. [PubMed] [Google Scholar]
  18. Leonard W. J., Depper J. M., Crabtree G. R., Rudikoff S., Pumphrey J., Robb R. J., Krönke M., Svetlik P. B., Peffer N. J., Waldmann T. A. Molecular cloning and expression of cDNAs for the human interleukin-2 receptor. Nature. 1984 Oct 18;311(5987):626–631. doi: 10.1038/311626a0. [DOI] [PubMed] [Google Scholar]
  19. Merida I., Gaulton G. N. Protein tyrosine phosphorylation associated with activation of the interleukin 2 receptor. J Biol Chem. 1990 Apr 5;265(10):5690–5694. [PubMed] [Google Scholar]
  20. Mills G. B., May C., McGill M., Fung M., Baker M., Sutherland R., Greene W. C. Interleukin 2-induced tyrosine phosphorylation. Interleukin 2 receptor beta is tyrosine phosphorylated. J Biol Chem. 1990 Feb 25;265(6):3561–3567. [PubMed] [Google Scholar]
  21. Morla A. O., Schreurs J., Miyajima A., Wang J. Y. Hematopoietic growth factors activate the tyrosine phosphorylation of distinct sets of proteins in interleukin-3-dependent murine cell lines. Mol Cell Biol. 1988 May;8(5):2214–2218. doi: 10.1128/mcb.8.5.2214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morrison D. K., Kaplan D. R., Escobedo J. A., Rapp U. R., Roberts T. M., Williams L. T. Direct activation of the serine/threonine kinase activity of Raf-1 through tyrosine phosphorylation by the PDGF beta-receptor. Cell. 1989 Aug 25;58(4):649–657. doi: 10.1016/0092-8674(89)90100-1. [DOI] [PubMed] [Google Scholar]
  23. Morrison D. K., Kaplan D. R., Rapp U., Roberts T. M. Signal transduction from membrane to cytoplasm: growth factors and membrane-bound oncogene products increase Raf-1 phosphorylation and associated protein kinase activity. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8855–8859. doi: 10.1073/pnas.85.23.8855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nemeth S. P., Fox L. G., DeMarco M., Brugge J. S. Deletions within the amino-terminal half of the c-src gene product that alter the functional activity of the protein. Mol Cell Biol. 1989 Mar;9(3):1109–1119. doi: 10.1128/mcb.9.3.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nikaido T., Shimizu A., Ishida N., Sabe H., Teshigawara K., Maeda M., Uchiyama T., Yodoi J., Honjo T. Molecular cloning of cDNA encoding human interleukin-2 receptor. Nature. 1984 Oct 18;311(5987):631–635. doi: 10.1038/311631a0. [DOI] [PubMed] [Google Scholar]
  26. Rapp U. R., Cleveland J. L., Brightman K., Scott A., Ihle J. N. Abrogation of IL-3 and IL-2 dependence by recombinant murine retroviruses expressing v-myc oncogenes. Nature. 1985 Oct 3;317(6036):434–438. doi: 10.1038/317434a0. [DOI] [PubMed] [Google Scholar]
  27. Reed J. C., Tadmori W., Kamoun M., Koretzky G., Nowell P. C. Suppression of interleukin 2 receptor acquisition by monoclonal antibodies recognizing the 50 KD protein associated with the sheep erythrocyte receptor on human T lymphocytes. J Immunol. 1985 Mar;134(3):1631–1639. [PubMed] [Google Scholar]
  28. Rosenberg S. A., Grimm E. A., McGrogan M., Doyle M., Kawasaki E., Koths K., Mark D. F. Biological activity of recombinant human interleukin-2 produced in Escherichia coli. Science. 1984 Mar 30;223(4643):1412–1414. doi: 10.1126/science.6367046. [DOI] [PubMed] [Google Scholar]
  29. Saltzman E. M., Thom R. R., Casnellie J. E. Activation of a tyrosine protein kinase is an early event in the stimulation of T lymphocytes by interleukin-2. J Biol Chem. 1988 May 25;263(15):6956–6959. [PubMed] [Google Scholar]
  30. Schultz A. M., Copeland T. D., Mark G. E., Rapp U. R., Oroszlan S. Detection of the myristylated gag-raf transforming protein with raf-specific antipeptide sera. Virology. 1985 Oct 15;146(1):78–89. doi: 10.1016/0042-6822(85)90054-6. [DOI] [PubMed] [Google Scholar]
  31. Severinson E., Naito T., Tokumoto H., Fukushima D., Hirano A., Hama K., Honjo T. Interleukin 4 (IgG1 induction factor): a multifunctional lymphokine acting also on T cells. Eur J Immunol. 1987 Jan;17(1):67–72. doi: 10.1002/eji.1830170112. [DOI] [PubMed] [Google Scholar]
  32. Sharon M., Gnarra J. R., Leonard W. J. The beta-chain of the IL-2 receptor (p70) is tyrosine-phosphorylated on YT and HUT-102B2 cells. J Immunol. 1989 Oct 15;143(8):2530–2533. [PubMed] [Google Scholar]
  33. Tsudo M., Karasuyama H., Kitamura F., Nagasaka Y., Tanaka T., Miyasaka M. Reconstitution of a functional IL-2 receptor by the beta-chain cDNA. A newly acquired receptor transduces negative signal. J Immunol. 1989 Dec 15;143(12):4039–4043. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES