Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 Oct 1;303(Pt 1):27–31. doi: 10.1042/bj3030027

The mechanism by which epidermal growth factor inhibits glycogen synthase kinase 3 in A431 cells.

Y Saito 1, J R Vandenheede 1, P Cohen 1
PMCID: PMC1137551  PMID: 7945252

Abstract

Glycogen synthase kinase 3 (GSK3) was inhibited by 50% within 5 min when A431 cells were stimulated with epidermal growth factor (EGF). The inhibition was unaffected by rapamycin at concentrations which blocked the activation of p70 S6 kinase, and reversed by incubation with protein phosphatase-1. EGF stimulation of A431 cells inhibited GSK3 alpha and GSK3 beta to a similar extent, and inhibition was accompanied by phosphorylation of the tryptic peptides containing the serine residues phosphorylated in vitro by p70 S6 kinase or MAP kinase-activated protein (MAPKAP) kinase-1 beta (also termed Rsk-2). These results demonstrate that EGF inhibits GSK3 by inducing phosphorylation of a serine residue and that GSK3 is not phosphorylated in vivo by either p70 S6 kinase or protein kinase C.

Full text

PDF
27

Images in this article

29Figure 2
on p.29
29Figure 3
on p.29

Selected References

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

  1. Alessi D. R., Street A. J., Cohen P., Cohen P. T. Inhibitor-2 functions like a chaperone to fold three expressed isoforms of mammalian protein phosphatase-1 into a conformation with the specificity and regulatory properties of the native enzyme. Eur J Biochem. 1993 May 1;213(3):1055–1066. doi: 10.1111/j.1432-1033.1993.tb17853.x. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Chung J., Kuo C. J., Crabtree G. R., Blenis J. Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases. Cell. 1992 Jun 26;69(7):1227–1236. doi: 10.1016/0092-8674(92)90643-q. [DOI] [PubMed] [Google Scholar]
  4. Cohen P., Alemany S., Hemmings B. A., Resink T. J., Strålfors P., Tung H. Y. Protein phosphatase-1 and protein phosphatase-2A from rabbit skeletal muscle. Methods Enzymol. 1988;159:390–408. doi: 10.1016/0076-6879(88)59039-0. [DOI] [PubMed] [Google Scholar]
  5. Cross D. A., Alessi D. R., Vandenheede J. R., McDowell H. E., Hundal H. S., Cohen P. The inhibition of glycogen synthase kinase-3 by insulin or insulin-like growth factor 1 in the rat skeletal muscle cell line L6 is blocked by wortmannin, but not by rapamycin: evidence that wortmannin blocks activation of the mitogen-activated protein kinase pathway in L6 cells between Ras and Raf. Biochem J. 1994 Oct 1;303(Pt 1):21–26. doi: 10.1042/bj3030021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Embi N., Rylatt D. B., Cohen P. Glycogen synthase kinase-3 from rabbit skeletal muscle. Separation from cyclic-AMP-dependent protein kinase and phosphorylase kinase. Eur J Biochem. 1980 Jun;107(2):519–527. [PubMed] [Google Scholar]
  7. Hardie D. G., Guy P. S. Reversible phosphorylation and inactivation of acetyl-CoA carboxylase from lactating rat mammary gland by cyclic AMP-dependent protein kinase. Eur J Biochem. 1980 Sep;110(1):167–177. doi: 10.1111/j.1432-1033.1980.tb04852.x. [DOI] [PubMed] [Google Scholar]
  8. Hughes K., Nikolakaki E., Plyte S. E., Totty N. F., Woodgett J. R. Modulation of the glycogen synthase kinase-3 family by tyrosine phosphorylation. EMBO J. 1993 Feb;12(2):803–808. doi: 10.1002/j.1460-2075.1993.tb05715.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hughes K., Ramakrishna S., Benjamin W. B., Woodgett J. R. Identification of multifunctional ATP-citrate lyase kinase as the alpha-isoform of glycogen synthase kinase-3. Biochem J. 1992 Nov 15;288(Pt 1):309–314. doi: 10.1042/bj2880309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Plyte S. E., Hughes K., Nikolakaki E., Pulverer B. J., Woodgett J. R. Glycogen synthase kinase-3: functions in oncogenesis and development. Biochim Biophys Acta. 1992 Dec 16;1114(2-3):147–162. doi: 10.1016/0304-419x(92)90012-n. [DOI] [PubMed] [Google Scholar]
  11. Ramakrishna S., Benjamin W. B. Insulin action rapidly decreases multifunctional protein kinase activity in rat adipose tissue. J Biol Chem. 1988 Sep 5;263(25):12677–12681. [PubMed] [Google Scholar]
  12. Ruel L., Bourouis M., Heitzler P., Pantesco V., Simpson P. Drosophila shaggy kinase and rat glycogen synthase kinase-3 have conserved activities and act downstream of Notch. Nature. 1993 Apr 8;362(6420):557–560. doi: 10.1038/362557a0. [DOI] [PubMed] [Google Scholar]
  13. Siegfried E., Perkins L. A., Capaci T. M., Perrimon N. Putative protein kinase product of the Drosophila segment-polarity gene zeste-white3. Nature. 1990 Jun 28;345(6278):825–829. doi: 10.1038/345825a0. [DOI] [PubMed] [Google Scholar]
  14. Sutherland C., Cohen P. The alpha-isoform of glycogen synthase kinase-3 from rabbit skeletal muscle is inactivated by p70 S6 kinase or MAP kinase-activated protein kinase-1 in vitro. FEBS Lett. 1994 Jan 24;338(1):37–42. doi: 10.1016/0014-5793(94)80112-6. [DOI] [PubMed] [Google Scholar]
  15. Sutherland C., Leighton I. A., Cohen P. Inactivation of glycogen synthase kinase-3 beta by phosphorylation: new kinase connections in insulin and growth-factor signalling. Biochem J. 1993 Nov 15;296(Pt 1):15–19. doi: 10.1042/bj2960015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Van Lint J., Khandelwal R. L., Merlevede W., Vandenheede J. R. A specific immunoprecipitation assay for the protein kinase FA/glycogen synthase kinase 3. Anal Biochem. 1993 Jan;208(1):132–137. doi: 10.1006/abio.1993.1018. [DOI] [PubMed] [Google Scholar]
  17. Wang Q. M., Fiol C. J., DePaoli-Roach A. A., Roach P. J. Glycogen synthase kinase-3 beta is a dual specificity kinase differentially regulated by tyrosine and serine/threonine phosphorylation. J Biol Chem. 1994 May 20;269(20):14566–14574. [PubMed] [Google Scholar]
  18. Welsh G. I., Foulstone E. J., Young S. W., Tavaré J. M., Proud C. G. Wortmannin inhibits the effects of insulin and serum on the activities of glycogen synthase kinase-3 and mitogen-activated protein kinase. Biochem J. 1994 Oct 1;303(Pt 1):15–20. doi: 10.1042/bj3030015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Welsh G. I., Proud C. G. Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B. Biochem J. 1993 Sep 15;294(Pt 3):625–629. doi: 10.1042/bj2940625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yu J. S., Yang S. D. Okadaic acid, a serine/threonine phosphatase inhibitor, induces tyrosine dephosphorylation/inactivation of protein kinase FA/GSK-3 alpha in A431 cells. J Biol Chem. 1994 May 20;269(20):14341–14344. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES