, Volume 8, Issue 2, pp 193–200 | Cite as

Insulin induces tyrosine phosphorylation of Shc and stimulates Shc/GRB2 association in insulin-sensitive tissues of the intact rat

  • Verónica Páez-Espinosa
  • Carla R. O. Carvalho
  • Fernanda Alvarez-Rojas
  • Luis Janeri
  • Licio A. Velloso
  • Antonio C. Boschero
  • Mario J. A. Saad


Shc is a novel type of tyrosine-phosphorylated protein activated in response to a wide variety of polypeptide ligands. In this study, we used immunoprecipitation and immunoblotting to examine the effect of insulin on Shc tyrosine phosphorylation and Shc/GRB2 association in insulin-sensitive tissues of the intact rat. Following an infusion of insulin, Shc was tyrosine-phosphorylated in the liver, skeletal muscle, and adipose tissue in a time- and dose-dependent fashion, which peaked 5 min after exposure to the hormone and, except in the case of adipose tissue, returned to basal values after 15 min. There was coimmunoprecipitation of Shc and the insulin receptor after stimulation with insulin. Receptor tyrosine kinase activity toward Shc was also observed. Following an infusion of insulin, Shc was found to associate with GRB2. These results demonstrate that after stimulation of rat tissues with insulin, Shc binds to the insulin receptor, is tyrosine-phosphorylated, and subsequently associates with GRB2.

Key Words

Shc GRB2 tyrosine phosphorylation tyrosine kinase activity 


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  1. 1.
    Kasuga, M., Hedo, J. A., Yamada, K. M., and Kahn, C. R. (1982). J. Biol. Chem. 257, 10,392–10,399.Google Scholar
  2. 2.
    White, M. F. and Kahn, C. R. (1989). J. Cell. Biochem. 39, 429–441.PubMedCrossRefGoogle Scholar
  3. 3.
    Fratalli, A. L., Treadway, J. L., and Pessin, J. E. (1992). J. Biol. Chem. 267, 19,521–19,528.Google Scholar
  4. 4.
    Myers, M. G., Jr. and White, M. F. (1993). Diabetes 42, 643–650.PubMedCrossRefGoogle Scholar
  5. 5.
    White, M. F. and Kahn, C. R. (1994), J. Biol. Chem. 269, 1–4.PubMedGoogle Scholar
  6. 6.
    White, M. F., Maron, R., and Khan, C. R. (1985). Nature 318, 183–186.PubMedCrossRefGoogle Scholar
  7. 7.
    Sun, X. J., Rothenberg, P. A., Kahn, C. R., Backer, J. M., Araki, E. Wilden, P. A., et al. (1991). Nature 352, 73–77.PubMedCrossRefGoogle Scholar
  8. 8.
    Araki, E., Sun, X. J., Haag, B. L., Chuang, L. M., Yang-Feng, T., White, M. F., et al. (1993). Diabetes 42, 1041–1054.PubMedCrossRefGoogle Scholar
  9. 9.
    Keller, S. R., Aebersold, R., Gardner, C. W., and Lienhard, G. E. (1993). Biochem. Biophys. Acta. 1172, 323–326.PubMedGoogle Scholar
  10. 10.
    Sun, X. J., Wang, L. M., Zhang, Y., Yenush, L. P, Myers, M. G., Glasheen, E., et al. (1995). Nature 377, 173–177.PubMedCrossRefGoogle Scholar
  11. 11.
    Pelicci, G., Lanfrancone, L., Grinani, F., McGlade, J., Cavallo, F., Forni, G., et al. (1992). Cell 70, 93–104.PubMedCrossRefGoogle Scholar
  12. 12.
    Burns, L. A., Karnitz, L. M., Sutor, S. L., and Abraham, R. T. (1993). J. Biol. Chem. 268, 17,659–17,661.Google Scholar
  13. 13.
    Giorgetti, S., Pelicci, P. G., Pelicci, G., and Van Obberghen, E. (1994). Eur. J. Biochem. 223, 195–202.PubMedCrossRefGoogle Scholar
  14. 14.
    Okada, S., Yamauchi, K., and Pessin, J. E. (1995). J. Biol. Chem. 270, 20,737–20,741.Google Scholar
  15. 15.
    Pronk, G. J., McGlade, J., Pelicci, G., Pawson, T., and Bos, J. L. (1993). J. Biol. Chem. 268, 5748–5753.PubMedGoogle Scholar
  16. 16.
    Lowestein, E. J., Daly, R. J., Batzer, A. G., Li, W., Margolis, B., Lammers, R., et al. (1992). Cell 70, 431–442.CrossRefGoogle Scholar
  17. 17.
    Mc Glade, J., Cheng, A., Pelicci, G., Pelicci, P. G., and Pawson, T. (1992). Proc. Natl. Acad. Sci. USA 89, 8869–8873.CrossRefGoogle Scholar
  18. 18.
    Rozakis-Adock, M., McGlade, J., Mbamalu, G., Pelicci, G., Daly, R., Li, W., et al. (1992). Nature 360, 689–692.CrossRefGoogle Scholar
  19. 19.
    Waters, S. B., Chen, D., Kao, A. W., Okada, S. Holt, K. H., and Pessin, J. E. (1996). J. Biol. Chem. 271, 18,224–18,230.Google Scholar
  20. 20.
    Simon, M. A., Botwell, D. D. L., Dodson, G. S., Laverty, T. R., and Rubin, G. M. (1991). Cell 67, 701–716.PubMedCrossRefGoogle Scholar
  21. 21.
    Martegani, E., Vanoni, M., Zippel, R., Coccetti, P., Brambilla, R., Ferrari, C., et al. (1992). EMBO J. 11, 2151–2157.PubMedGoogle Scholar
  22. 22.
    Skolnik, E. Y., Batzer, A., Li, N., Lee, C. H., Lowestein, E. J., Mohammadi, M., et al. (1993). Science. 260, 1953–1955.PubMedCrossRefGoogle Scholar
  23. 23.
    Gustafson, T. A., He, W., Craparo, A., Schaub, C. D., and O’Neill, T. J. (1995). Mol. Cell. Biol. 15, 2500–2508.PubMedGoogle Scholar
  24. 24.
    Sasaoka, T., Draznin, B., Leitner, J. W., Langlois, W. J., and Olefsky, J. M. (1994). J. Biol. Chem. 269, 10,734–10,738.Google Scholar
  25. 25.
    Sasaoka, T., Langlois, W. J., Leitner, J. W., and Olefsky, J. M. (1994). J. Biol. Chem. 269, 32,621–32,625.Google Scholar
  26. 26.
    Pawson, T. and Gish, G. D. (1992). Cell 71, 359–362.PubMedCrossRefGoogle Scholar
  27. 27.
    Schlessinger, J. and Ullrich, A. (1992). Neuron 9, 383–391.PubMedCrossRefGoogle Scholar
  28. 28.
    Fantl, W. J., Johnson, D. E., and Williams, L. T. (1993). Annu. Rev. Biochem. 62, 453–481.PubMedGoogle Scholar
  29. 29.
    Mayer, B. J. and Baltimore, D. (1993). Trends. Cell. Biol. 3, 8–13.PubMedCrossRefGoogle Scholar
  30. 30.
    van der Geer, P. and Hunter, T. (1994). Annu. Rev. Cell. Biol. 10, 251–337.PubMedCrossRefGoogle Scholar
  31. 31.
    Di Guglielmo, G. M., Baass, P. C., Ou, W., Posner, B. I., and Bergeron, J. J. M. (1994). EMBO J. 13, 4269–4277.PubMedGoogle Scholar
  32. 32.
    Tartare-Deckert, S., Sawka-Verhelle, D., Murdaca, J., and Van Obberghen, E. (1995). J. Biol. Chem. 270, 23,456–23,460.Google Scholar
  33. 33.
    Segatto, O., Pelicci, G., Giuli, S., Digiesi, G., Di Fiore, P. P., McGlade, J., Pawson, T., et al. (1993). Oncogene 8, 2105–2112.PubMedGoogle Scholar
  34. 34.
    Obermeier, A., Bradshaw, R. A., Seedorf, K., Choidas, A., Schlessinger, J. and Ullrich, A. (1994). EMBO. 13, 1585–1590.Google Scholar
  35. 35.
    Craparo, A., O’Neill, T. J., and Gustafson, A. (1995). J. Biol. Chem. 270, 15,639–15,643.Google Scholar
  36. 36.
    Bork, P. and Margolis, B. (1995). Cell 80, 693–694.PubMedCrossRefGoogle Scholar
  37. 37.
    Batzer, A. G., Blaikie, P., Nelson, K., Schlessinger, J., and Margolis, B. (1995). Mol. Cell. Biol. 15, 4403–4409.PubMedGoogle Scholar
  38. 38.
    van der Geer, P., Willey, S., Lai, V. K. M., Olivier, J. P., Gish, G. D., Stephens, R., et al. (1996). Curr. Biol. 5, 404–412.Google Scholar
  39. 39.
    Isakoff, S. J., Yu, Y. P., Su, Y. Ch., Blaikie, P., Yajnik, V., Rose, E., et al. (1996). J. Biol. Chem. 271, 3959–3962.PubMedCrossRefGoogle Scholar
  40. 40.
    Carvalho, C. R. O., Brenelli, S. L., Silva, A. C., Nunes, A. L. B., Velloso, L. A., and Saad, M. J. A. (1996). Endocrinology 137, 151–159.PubMedCrossRefGoogle Scholar
  41. 41.
    Saad, M. J. A., Araki, E., Miralpeix, M., Rothenberg, O. L., White, M. F., and Kahn, C. R. (1992). J. Clin. Invest. 90, 1839–1849.PubMedGoogle Scholar
  42. 42.
    Ouwens, D. M., van der Zon, G. C. M., Pronk, G. J., Bos, J. L., Moller, W., Cheatam, B., et al. (1994). J. Biol. Chem. 269, 33,116–33,122.Google Scholar
  43. 43.
    Yonezawa, K., Ando, A., Kaburagi, Y., Yamamoto-Honda, R., Kitamura, T., Hara, K., et al. (1994). J. Biol. Chem. 269, 4634–4640.PubMedGoogle Scholar
  44. 44.
    Giorgino, F. and Smith, R. J. (1995). J. Clin. Invest. 96, 1473–1483.PubMedCrossRefGoogle Scholar
  45. 45.
    Li, P. M. and Goldstein, B. J. (1996). Biochem. Biophys. Res. Commun. 223, 80–84.PubMedCrossRefGoogle Scholar
  46. 46.
    Towbin, H., Staehlin, J., and Gordon, J. (1979). Proc. Natl. Acad. Sci. USA 76, 4350–4354.PubMedCrossRefGoogle Scholar
  47. 47.
    Laemmli, U. K. (1970). Nature 227, 680–685.PubMedCrossRefGoogle Scholar
  48. 48.
    Saad, M. J. A., Carvalho, C. R. O., Thirone, A. C. P., and Velloso, L. A. (1996). J. Biol. Chem. 271, 22,100–22,104.Google Scholar
  49. 49.
    Sokal, R. R. and Rohlf, F. J. (eds.) (1981). Biometry. W. H. Freeman and Company, New York.Google Scholar

Copyright information

© Humana Press Inc 1998

Authors and Affiliations

  • Verónica Páez-Espinosa
    • 1
  • Carla R. O. Carvalho
    • 2
  • Fernanda Alvarez-Rojas
    • 1
  • Luis Janeri
    • 2
  • Licio A. Velloso
    • 2
  • Antonio C. Boschero
    • 1
  • Mario J. A. Saad
    • 2
  1. 1.Departamento de Fisiologia e Biofísica, Instituto de BiologiaUNICAMPCampinas, SPBrasil
  2. 2.Departamento de Clínica MédicaFCM-UNICAMPCampinas, SPBrasil

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