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. 2002 Aug 15;366(Pt 1):57–62. doi: 10.1042/BJ20020198

The Src-family tyrosine kinase inhibitor PP1 interferes with the activation of ribosomal protein S6 kinases.

O Jameel Shah 1, Scot R Kimball 1, Leonard S Jefferson 1
PMCID: PMC1222761  PMID: 12014987

Abstract

Considerable biochemical and pharmacological evidence suggests that the activation of ribosomal protein S6 kinases (S6Ks) by activated receptor tyrosine kinases involves multiple co-ordinated input signals. However, the identities of many of these inputs remain poorly described, and their precise involvement in S6K activation has been the subject of great investigative effort. In the present study, we have shown that 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP1), a selective inhibitor of the Src family of non-receptor tyrosine kinases, interferes with the activation of 70 and 85 kDa S6K gene products (p70S6K1 and p85S6K1) by insulin, insulin-like growth factor 1, sodium orthovanadate and activated alleles of phosphoinositide 3-kinase and H-Ras. PP1 also impedes the activation of AKT/protein kinase B and the extracellular signal-regulated protein kinases 1 and 2 by these various stimuli. Insulin-like growth factor 1 was observed to induce a sustained increase in c-Src autophosphorylation as revealed using anti-phospho-Y416 antisera, but this effect was absent from the cells treated with PP1. To conclude, an activated allele of p70S6K1 is compared with the wild-type allele, resistant to inhibition by PP1 when co-expressed with phosphoinositide-dependent kinase 1 (PDK1), suggesting that PP1 affects p70S6K1 via a PDK1-independent pathway. Thus activation of Src may supply a necessary signal for the activation of p70S6K1 and possibly other S6Ks.

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Selected References

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  1. Akimoto K., Nakaya M., Yamanaka T., Tanaka J., Matsuda S., Weng Q. P., Avruch J., Ohno S. Atypical protein kinase Clambda binds and regulates p70 S6 kinase. Biochem J. 1998 Oct 15;335(Pt 2):417–424. doi: 10.1042/bj3350417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alessi D. R., Kozlowski M. T., Weng Q. P., Morrice N., Avruch J. 3-Phosphoinositide-dependent protein kinase 1 (PDK1) phosphorylates and activates the p70 S6 kinase in vivo and in vitro. Curr Biol. 1998 Jan 15;8(2):69–81. doi: 10.1016/s0960-9822(98)70037-5. [DOI] [PubMed] [Google Scholar]
  3. Ballif B. A., Shimamura A., Pae E., Blenis J. Disruption of 3-phosphoinositide-dependent kinase 1 (PDK1) signaling by the anti-tumorigenic and anti-proliferative agent n-alpha-tosyl-l-phenylalanyl chloromethyl ketone. J Biol Chem. 2001 Jan 18;276(15):12466–12475. doi: 10.1074/jbc.M009939200. [DOI] [PubMed] [Google Scholar]
  4. Bar-Sagi D., Hall A. Ras and Rho GTPases: a family reunion. Cell. 2000 Oct 13;103(2):227–238. doi: 10.1016/s0092-8674(00)00115-x. [DOI] [PubMed] [Google Scholar]
  5. Chou M. M., Blenis J. The 70 kDa S6 kinase complexes with and is activated by the Rho family G proteins Cdc42 and Rac1. Cell. 1996 May 17;85(4):573–583. doi: 10.1016/s0092-8674(00)81257-x. [DOI] [PubMed] [Google Scholar]
  6. Datta K., Bellacosa A., Chan T. O., Tsichlis P. N. Akt is a direct target of the phosphatidylinositol 3-kinase. Activation by growth factors, v-src and v-Ha-ras, in Sf9 and mammalian cells. J Biol Chem. 1996 Nov 29;271(48):30835–30839. doi: 10.1074/jbc.271.48.30835. [DOI] [PubMed] [Google Scholar]
  7. Dennis P. B., Pullen N., Kozma S. C., Thomas G. The principal rapamycin-sensitive p70(s6k) phosphorylation sites, T-229 and T-389, are differentially regulated by rapamycin-insensitive kinase kinases. Mol Cell Biol. 1996 Nov;16(11):6242–6251. doi: 10.1128/mcb.16.11.6242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dufner A., Thomas G. Ribosomal S6 kinase signaling and the control of translation. Exp Cell Res. 1999 Nov 25;253(1):100–109. doi: 10.1006/excr.1999.4683. [DOI] [PubMed] [Google Scholar]
  9. Edelmann H. M., Kühne C., Petritsch C., Ballou L. M. Cell cycle regulation of p70 S6 kinase and p42/p44 mitogen-activated protein kinases in Swiss mouse 3T3 fibroblasts. J Biol Chem. 1996 Jan 12;271(2):963–971. doi: 10.1074/jbc.271.2.963. [DOI] [PubMed] [Google Scholar]
  10. Grammer T. C., Cheatham L., Chou M. M., Blenis J. The p70S6K signalling pathway: a novel signalling system involved in growth regulation. Cancer Surv. 1996;27:271–292. [PubMed] [Google Scholar]
  11. Hanke J. H., Gardner J. P., Dow R. L., Changelian P. S., Brissette W. H., Weringer E. J., Pollok B. A., Connelly P. A. Discovery of a novel, potent, and Src family-selective tyrosine kinase inhibitor. Study of Lck- and FynT-dependent T cell activation. J Biol Chem. 1996 Jan 12;271(2):695–701. doi: 10.1074/jbc.271.2.695. [DOI] [PubMed] [Google Scholar]
  12. Imamoto A., Soriano P. Disruption of the csk gene, encoding a negative regulator of Src family tyrosine kinases, leads to neural tube defects and embryonic lethality in mice. Cell. 1993 Jun 18;73(6):1117–1124. doi: 10.1016/0092-8674(93)90641-3. [DOI] [PubMed] [Google Scholar]
  13. Karnitz L. M., Sutor S. L., Abraham R. T. The Src-family kinase, Fyn, regulates the activation of phosphatidylinositol 3-kinase in an interleukin 2-responsive T cell line. J Exp Med. 1994 Jun 1;179(6):1799–1808. doi: 10.1084/jem.179.6.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kimball S. R., Horetsky R. L., Jefferson L. S. Signal transduction pathways involved in the regulation of protein synthesis by insulin in L6 myoblasts. Am J Physiol. 1998 Jan;274(1 Pt 1):C221–C228. doi: 10.1152/ajpcell.1998.274.1.C221. [DOI] [PubMed] [Google Scholar]
  15. Kimball S. R., Karinch A. M., Feldhoff R. C., Mellor H., Jefferson L. S. Purification and characterization of eukaryotic translational initiation factor eIF-2B from liver. Biochim Biophys Acta. 1994 Dec 15;1201(3):473–481. doi: 10.1016/0304-4165(94)90079-5. [DOI] [PubMed] [Google Scholar]
  16. Liu A. X., Testa J. R., Hamilton T. C., Jove R., Nicosia S. V., Cheng J. Q. AKT2, a member of the protein kinase B family, is activated by growth factors, v-Ha-ras, and v-src through phosphatidylinositol 3-kinase in human ovarian epithelial cancer cells. Cancer Res. 1998 Jul 15;58(14):2973–2977. [PubMed] [Google Scholar]
  17. Liu X., Marengere L. E., Koch C. A., Pawson T. The v-Src SH3 domain binds phosphatidylinositol 3'-kinase. Mol Cell Biol. 1993 Sep;13(9):5225–5232. doi: 10.1128/mcb.13.9.5225-5232.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mak P., He Z., Kurosaki T. Identification of amino acid residues required for a specific interaction between Src-tyrosine kinase and proline-rich region of phosphatidylinositol-3' kinase. FEBS Lett. 1996 Nov 18;397(2-3):183–185. doi: 10.1016/s0014-5793(96)01179-9. [DOI] [PubMed] [Google Scholar]
  19. Nada S., Yagi T., Takeda H., Tokunaga T., Nakagawa H., Ikawa Y., Okada M., Aizawa S. Constitutive activation of Src family kinases in mouse embryos that lack Csk. Cell. 1993 Jun 18;73(6):1125–1135. doi: 10.1016/0092-8674(93)90642-4. [DOI] [PubMed] [Google Scholar]
  20. Papst P. J., Sugiyama H., Nagasawa M., Lucas J. J., Maller J. L., Terada N. Cdc2-cyclin B phosphorylates p70 S6 kinase on Ser411 at mitosis. J Biol Chem. 1998 Jun 12;273(24):15077–15084. doi: 10.1074/jbc.273.24.15077. [DOI] [PubMed] [Google Scholar]
  21. Pearson R. B., Dennis P. B., Han J. W., Williamson N. A., Kozma S. C., Wettenhall R. E., Thomas G. The principal target of rapamycin-induced p70s6k inactivation is a novel phosphorylation site within a conserved hydrophobic domain. EMBO J. 1995 Nov 1;14(21):5279–5287. doi: 10.1002/j.1460-2075.1995.tb00212.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Penuel E., Martin G. S. Transformation by v-Src: Ras-MAPK and PI3K-mTOR mediate parallel pathways. Mol Biol Cell. 1999 Jun;10(6):1693–1703. doi: 10.1091/mbc.10.6.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pullen N., Dennis P. B., Andjelkovic M., Dufner A., Kozma S. C., Hemmings B. A., Thomas G. Phosphorylation and activation of p70s6k by PDK1. Science. 1998 Jan 30;279(5351):707–710. doi: 10.1126/science.279.5351.707. [DOI] [PubMed] [Google Scholar]
  24. Rodriguez-Viciana P., Warne P. H., Vanhaesebroeck B., Waterfield M. D., Downward J. Activation of phosphoinositide 3-kinase by interaction with Ras and by point mutation. EMBO J. 1996 May 15;15(10):2442–2451. [PMC free article] [PubMed] [Google Scholar]
  25. Romanelli A., Martin K. A., Toker A., Blenis J. p70 S6 kinase is regulated by protein kinase Czeta and participates in a phosphoinositide 3-kinase-regulated signalling complex. Mol Cell Biol. 1999 Apr;19(4):2921–2928. doi: 10.1128/mcb.19.4.2921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schwartz M. A., Baron V. Interactions between mitogenic stimuli, or, a thousand and one connections. Curr Opin Cell Biol. 1999 Apr;11(2):197–202. doi: 10.1016/s0955-0674(99)80026-x. [DOI] [PubMed] [Google Scholar]
  27. Shah O. J., Kimball S. R., Jefferson L. S. Among translational effectors, p70S6k is uniquely sensitive to inhibition by glucocorticoids. Biochem J. 2000 Apr 15;347(Pt 2):389–397. doi: 10.1042/0264-6021:3470389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thomas G. An encore for ribosome biogenesis in the control of cell proliferation. Nat Cell Biol. 2000 May;2(5):E71–E72. doi: 10.1038/35010581. [DOI] [PubMed] [Google Scholar]
  29. Wages D. S., Keefer J., Rall T. B., Weber M. J. Mutations in the SH3 domain of the src oncogene which decrease association of phosphatidylinositol 3'-kinase activity with pp60v-src and alter cellular morphology. J Virol. 1992 Apr;66(4):1866–1874. doi: 10.1128/jvi.66.4.1866-1874.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weng Q. P., Andrabi K., Klippel A., Kozlowski M. T., Williams L. T., Avruch J. Phosphatidylinositol 3-kinase signals activation of p70 S6 kinase in situ through site-specific p70 phosphorylation. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5744–5748. doi: 10.1073/pnas.92.12.5744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Whitman M., Downes C. P., Keeler M., Keller T., Cantley L. Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate. Nature. 1988 Apr 14;332(6165):644–646. doi: 10.1038/332644a0. [DOI] [PubMed] [Google Scholar]
  32. Wooten M. W., Seibenhener M. L., Neidigh K. B., Vandenplas M. L. Mapping of atypical protein kinase C within the nerve growth factor signaling cascade: relationship to differentiation and survival of PC12 cells. Mol Cell Biol. 2000 Jul;20(13):4494–4504. doi: 10.1128/mcb.20.13.4494-4504.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]

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