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. 1995 Aug 1;182(2):593–597. doi: 10.1084/jem.182.2.593

Insulin-like growth factor induces phosphorylation of immunoreactive insulin receptor substrate and its association with phosphatidylinositol-3 kinase in human thymocytes

PMCID: PMC2192136  PMID: 7543144

Abstract

Insulin receptor substrate 1 (IRS-1) is the principle cellular substrate for insulin and insulin-like growth factor I (IGF-I) receptor signaling. After phosphorylation of tyrosine residues within the YMXM or YXXM motifs, IRS-1 associates with phosphatidylinositol-3 kinase (PI3K). This signaling pathway and the presence of an IRS-1-like molecule have been demonstrated in IRS-1-transfected and in nontransfected hematopoietic cell lines, respectively. IGF-I has been implicated in lymphocyte development and function, and recently, we showed that functional type-I IGF receptors are present on human thymocytes and peripheral T cells. In this study, we addressed IGF-I signal transduction in nontransformed, freshly isolated, human thymocytes, as well as in blood T cells. Using Western blot analysis, we found that IGF-I induced phosphorylation of a 160-180-kD protein (pp170) in human thymocytes and that phosphorylated pp170 becomes associated with PI3K and is recognized by anti-IRS-1. In blood T cells, this immunoreactive IRS-1 (irIRS-1) is less abundantly expressed than in thymocytes, as assessed with immunoblotting. As a consequence, phosphorylated pp170 was not or hardly detectable after stimulation with IGF-I, and irIRS-1 was not detected in PI3K immunoprecipitates from lysates of IGF-I-stimulated T cells. However, IGF-I induced the tyrosine phosphorylation of other cellular proteins, indicating that differential expression of irIRS-1 contributes to a distinct signaling pathway in T cells.

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Selected References

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  1. Backer J. M., Myers M. G., Jr, Shoelson S. E., Chin D. J., Sun X. J., Miralpeix M., Hu P., Margolis B., Skolnik E. Y., Schlessinger J. Phosphatidylinositol 3'-kinase is activated by association with IRS-1 during insulin stimulation. EMBO J. 1992 Sep;11(9):3469–3479. doi: 10.1002/j.1460-2075.1992.tb05426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bajpai A., Brahmi Z. Regulation of resting and IL-2-activated human cytotoxic lymphocytes by exogenous nucleotides: role of IL-2 and ecto-ATPases. Cell Immunol. 1993 Apr 15;148(1):130–143. doi: 10.1006/cimm.1993.1096. [DOI] [PubMed] [Google Scholar]
  3. Clark R., Strasser J., McCabe S., Robbins K., Jardieu P. Insulin-like growth factor-1 stimulation of lymphopoiesis. J Clin Invest. 1993 Aug;92(2):540–548. doi: 10.1172/JCI116621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dadi H. K., Roifman C. M. Activation of phosphatidylinositol-3 kinase by ligation of the interleukin-7 receptor on human thymocytes. J Clin Invest. 1993 Sep;92(3):1559–1563. doi: 10.1172/JCI116736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gmelig-Meyling F., UytdeHaag A. G., Ballieux R. E. Human B-cell activation in vitro. T cell-dependent pokeweed mitogen-induced differentiation of blood B lymphocytes. Cell Immunol. 1977 Sep;33(1):156–169. doi: 10.1016/0008-8749(77)90143-5. [DOI] [PubMed] [Google Scholar]
  6. Hosomi Y., Shii K., Ogawa W., Matsuba H., Yoshida M., Okada Y., Yokono K., Kasuga M., Baba S., Roth R. A. Characterization of a 60-kilodalton substrate of the insulin receptor kinase. J Biol Chem. 1994 Apr 15;269(15):11498–11502. [PubMed] [Google Scholar]
  7. Izumi T., White M. F., Kadowaki T., Takaku F., Akanuma Y., Kasuga M. Insulin-like growth factor I rapidly stimulates tyrosine phosphorylation of a Mr 185,000 protein in intact cells. J Biol Chem. 1987 Jan 25;262(3):1282–1287. [PubMed] [Google Scholar]
  8. Johnson E. W., Jones L. A., Kozak R. W. Expression and function of insulin-like growth factor receptors on anti-CD3-activated human T lymphocytes. J Immunol. 1992 Jan 1;148(1):63–71. [PubMed] [Google Scholar]
  9. June C. H., Fletcher M. C., Ledbetter J. A., Schieven G. L., Siegel J. N., Phillips A. F., Samelson L. E. Inhibition of tyrosine phosphorylation prevents T-cell receptor-mediated signal transduction. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7722–7726. doi: 10.1073/pnas.87.19.7722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kelley K. W., Arkins S., Li Y. M. Growth hormone, prolactin, and insulin-like growth factors: new jobs for old players. Brain Behav Immun. 1992 Dec;6(4):317–326. [PubMed] [Google Scholar]
  11. Kooijman R. K., Scholtens L. E., Rijkers G. T., Zegers B. J. Differential expression of type I insulin-like growth factor receptors in different stages of human T cells. Eur J Immunol. 1995 Apr;25(4):931–935. doi: 10.1002/eji.1830250411. [DOI] [PubMed] [Google Scholar]
  12. Kooijman R., Willems M., De Haas C. J., Rijkers G. T., Schuurmans A. L., Van Buul-Offers S. C., Heijnen C. J., Zegers B. J. Expression of type I insulin-like growth factor receptors on human peripheral blood mononuclear cells. Endocrinology. 1992 Nov;131(5):2244–2250. doi: 10.1210/endo.131.5.1425423. [DOI] [PubMed] [Google Scholar]
  13. Kooijman R., Willems M., Rijkers G. T., Brinkman A., van Buul-Offers S. C., Heijnen C. J., Zegers B. J. Effects of insulin-like growth factors and growth hormone on the in vitro proliferation of T lymphocytes. J Neuroimmunol. 1992 May;38(1-2):95–104. doi: 10.1016/0165-5728(92)90094-2. [DOI] [PubMed] [Google Scholar]
  14. Kovacina K. S., Roth R. A. Identification of SHC as a substrate of the insulin receptor kinase distinct from the GAP-associated 62 kDa tyrosine phosphoprotein. Biochem Biophys Res Commun. 1993 May 14;192(3):1303–1311. doi: 10.1006/bbrc.1993.1558. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Landreth K. S., Narayanan R., Dorshkind K. Insulin-like growth factor-I regulates pro-B cell differentiation. Blood. 1992 Sep 1;80(5):1207–1212. [PubMed] [Google Scholar]
  17. Lavan B. E., Lienhard G. E. The insulin-elicited 60-kDa phosphotyrosine protein in rat adipocytes is associated with phosphatidylinositol 3-kinase. J Biol Chem. 1993 Mar 15;268(8):5921–5928. [PubMed] [Google Scholar]
  18. Margolis R. N., Schell M. J., Taylor S. I., Hubbard A. L. Hepatocyte plasma membrane ECTO-ATPase (pp120/HA4) is a substrate for tyrosine kinase activity of the insulin receptor. Biochem Biophys Res Commun. 1990 Jan 30;166(2):562–566. doi: 10.1016/0006-291x(90)90845-e. [DOI] [PubMed] [Google Scholar]
  19. Myers M. G., Jr, Sun X. J., Cheatham B., Jachna B. R., Glasheen E. M., Backer J. M., White M. F. IRS-1 is a common element in insulin and insulin-like growth factor-I signaling to the phosphatidylinositol 3'-kinase. Endocrinology. 1993 Apr;132(4):1421–1430. doi: 10.1210/endo.132.4.8384986. [DOI] [PubMed] [Google Scholar]
  20. Perrotti N., Accili D., Marcus-Samuels B., Rees-Jones R. W., Taylor S. I. Insulin stimulates phosphorylation of a 120-kDa glycoprotein substrate (pp120) for the receptor-associated protein kinase in intact H-35 hepatoma cells. Proc Natl Acad Sci U S A. 1987 May;84(10):3137–3140. doi: 10.1073/pnas.84.10.3137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Porras A., Nebreda A. R., Benito M., Santos E. Activation of Ras by insulin in 3T3 L1 cells does not involve GTPase-activating protein phosphorylation. J Biol Chem. 1992 Oct 15;267(29):21124–21131. [PubMed] [Google Scholar]
  22. Reiss K., Porcu P., Sell C., Pietrzkowski Z., Baserga R. The insulin-like growth factor 1 receptor is required for the proliferation of hemopoietic cells. Oncogene. 1992 Nov;7(11):2243–2248. [PubMed] [Google Scholar]
  23. Roth R. A., Zhang B., Chin J. E., Kovacina K. Substrates and signalling complexes: the tortured path to insulin action. J Cell Biochem. 1992 Jan;48(1):12–18. doi: 10.1002/jcb.240480104. [DOI] [PubMed] [Google Scholar]
  24. Samelson L. E., Fletcher M. C., Ledbetter J. A., June C. H. Activation of tyrosine phosphorylation in human T cells via the CD2 pathway. Regulation by the CD45 tyrosine phosphatase. J Immunol. 1990 Oct 15;145(8):2448–2454. [PubMed] [Google Scholar]
  25. Schimpff R. M., Repellin A. M., Salvatoni A., Thieriot-Prevost G., Chatelain P. Effect of purified somatomedins on thymidine incorporation into lectin-activated human lymphocytes. Acta Endocrinol (Copenh) 1983 Jan;102(1):21–26. doi: 10.1530/acta.0.1020021. [DOI] [PubMed] [Google Scholar]
  26. Shoelson S. E., Chatterjee S., Chaudhuri M., White M. F. YMXM motifs of IRS-1 define substrate specificity of the insulin receptor kinase. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2027–2031. doi: 10.1073/pnas.89.6.2027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sun X. J., Rothenberg P., Kahn C. R., Backer J. M., Araki E., Wilden P. A., Cahill D. A., Goldstein B. J., White M. F. Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein. Nature. 1991 Jul 4;352(6330):73–77. doi: 10.1038/352073a0. [DOI] [PubMed] [Google Scholar]
  28. Tapson V. F., Boni-Schnetzler M., Pilch P. F., Center D. M., Berman J. S. Structural and functional characterization of the human T lymphocyte receptor for insulin-like growth factor I in vitro. J Clin Invest. 1988 Sep;82(3):950–957. doi: 10.1172/JCI113703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ugi S., Maegawa H., Olefsky J. M., Shigeta Y., Kashiwagi A. Src homology 2 domains of protein tyrosine phosphatase are associated in vitro with both the insulin receptor and insulin receptor substrate-1 via different phosphotyrosine motifs. FEBS Lett. 1994 Mar 7;340(3):216–220. doi: 10.1016/0014-5793(94)80141-x. [DOI] [PubMed] [Google Scholar]
  31. Wang L. M., Keegan A. D., Li W., Lienhard G. E., Pacini S., Gutkind J. S., Myers M. G., Jr, Sun X. J., White M. F., Aaronson S. A. Common elements in interleukin 4 and insulin signaling pathways in factor-dependent hematopoietic cells. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4032–4036. doi: 10.1073/pnas.90.9.4032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wang L. M., Myers M. G., Jr, Sun X. J., Aaronson S. A., White M., Pierce J. H. IRS-1: essential for insulin- and IL-4-stimulated mitogenesis in hematopoietic cells. Science. 1993 Sep 17;261(5128):1591–1594. doi: 10.1126/science.8372354. [DOI] [PubMed] [Google Scholar]
  33. White M. F., Maron R., Kahn C. R. Insulin rapidly stimulates tyrosine phosphorylation of a Mr-185,000 protein in intact cells. Nature. 1985 Nov 14;318(6042):183–186. doi: 10.1038/318183a0. [DOI] [PubMed] [Google Scholar]
  34. White M. F., Stegmann E. W., Dull T. J., Ullrich A., Kahn C. R. Characterization of an endogenous substrate of the insulin receptor in cultured cells. J Biol Chem. 1987 Jul 15;262(20):9769–9777. [PubMed] [Google Scholar]

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