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. 1986 Dec;83(24):9294–9297. doi: 10.1073/pnas.83.24.9294

Multisite phosphorylation of the alpha subunit of transducin by the insulin receptor kinase and protein kinase C.

Y Zick, R Sagi-Eisenberg, M Pines, P Gierschik, A M Spiegel
PMCID: PMC387124  PMID: 3099281

Abstract

The GDP-bound alpha subunit of transducin, but not the guanosine 5'-[gamma-thio]triphosphate-bound one, undergoes phosphorylation on tyrosine residues by the insulin receptor kinase and on serine residues by protein kinase C. Holotransducin is poorly phosphorylated by the insulin receptor kinase and is not phosphorylated by protein kinase C. Neither holotransducin nor any of its subunits were phosphorylated by the cAMP-dependent protein kinase. That a given subunit of transducin undergoes multisite phosphorylation depending on the type of nucleotide bound to it or the nature of the kinase suggests that hormone-dependent phosphorylation could provide a versatile mode for regulation of guanine nucleotide-binding protein (G protein) function. In particular, the findings that certain G proteins serve as substrates for both the insulin receptor kinase and protein kinase C implicate G proteins in playing a key role in mediating the action of insulin and ligands that act to activate protein kinase C.

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Selected References

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

  1. Avruch J., Nemenoff R. A., Blackshear P. J., Pierce M. W., Osathanondh R. Insulin-stimulated tyrosine phosphorylation of the insulin receptor in detergent extracts of human placental membranes. Comparison to epidermal growth factor-stimulated phosphorylation. J Biol Chem. 1982 Dec 25;257(24):15162–15166. [PubMed] [Google Scholar]
  2. Bray G. A., York D. A. Genetically transmitted obesity in rodents. Physiol Rev. 1971 Jul;51(3):598–646. doi: 10.1152/physrev.1971.51.3.598. [DOI] [PubMed] [Google Scholar]
  3. Bégin-Heick N. Absence of the inhibitory effect of guanine nucleotides on adenylate cyclase activity in white adipocyte membranes of the ob/ob mouse. Effect of the ob gene. J Biol Chem. 1985 May 25;260(10):6187–6193. [PubMed] [Google Scholar]
  4. Cassel D., Selinger Z. Catecholamine-stimulated GTPase activity in turkey erythrocyte membranes. Biochim Biophys Acta. 1976 Dec 8;452(2):538–551. doi: 10.1016/0005-2744(76)90206-0. [DOI] [PubMed] [Google Scholar]
  5. Cassel D., Selinger Z. Mechanism of adenylate cyclase activation through the beta-adrenergic receptor: catecholamine-induced displacement of bound GDP by GTP. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4155–4159. doi: 10.1073/pnas.75.9.4155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  7. Elks M. L., Watkins P. A., Manganiello V. C., Moss J., Hewlett E., Vaughan M. Selective regulation by pertussis toxin of insulin-induced activation of particulate cAMP phosphodiesterase activity in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 1983 Oct 31;116(2):593–598. doi: 10.1016/0006-291x(83)90565-x. [DOI] [PubMed] [Google Scholar]
  8. Enomoto K., Gill D. M. Cholera toxin activation of adenylate cyclase. Roles of nucleoside triphosphates and a macromolecular factor in the ADP ribosylation of the GTP-dependent regulatory component. J Biol Chem. 1980 Feb 25;255(4):1252–1258. [PubMed] [Google Scholar]
  9. Farese R. V., Standaert M. L., Barnes D. E., Davis J. S., Pollet R. J. Phorbol ester provokes insulin-like effects on glucose transport, amino acid uptake, and pyruvate dehydrogenase activity in BC3H-1 cultured myocytes. Endocrinology. 1985 Jun;116(6):2650–2655. doi: 10.1210/endo-116-6-2650. [DOI] [PubMed] [Google Scholar]
  10. Fung B. K., Nash C. R. Characterization of transducin from bovine retinal rod outer segments. II. Evidence for distinct binding sites and conformational changes revealed by limited proteolysis with trypsin. J Biol Chem. 1983 Sep 10;258(17):10503–10510. [PubMed] [Google Scholar]
  11. Ghalayini A., Anderson R. E. Phosphatidylinositol 4,5-bisphosphate: light-mediated breakdown in the vertebrate retina. Biochem Biophys Res Commun. 1984 Oct 30;124(2):503–506. doi: 10.1016/0006-291x(84)91582-1. [DOI] [PubMed] [Google Scholar]
  12. Gierschik P., Codina J., Simons C., Birnbaumer L., Spiegel A. Antisera against a guanine nucleotide binding protein from retina cross-react with the beta subunit of the adenylyl cyclase-associated guanine nucleotide binding proteins, Ns and Ni. Proc Natl Acad Sci U S A. 1985 Feb;82(3):727–731. doi: 10.1073/pnas.82.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Graves C. B., McDonald J. M. Insulin and phorbol ester stimulate phosphorylation of a 40-kDa protein in adipocyte plasma membranes. J Biol Chem. 1985 Sep 15;260(20):11286–11292. [PubMed] [Google Scholar]
  14. Heyworth C. M., Whetton A. D., Wong S., Martin B. R., Houslay M. D. Insulin inhibits the cholera-toxin-catalysed ribosylation of a Mr-25000 protein in rat liver plasma membranes. Biochem J. 1985 Jun 15;228(3):593–603. doi: 10.1042/bj2280593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hildebrandt J. D., Sekura R. D., Codina J., Iyengar R., Manclark C. R., Birnbaumer L. Stimulation and inhibition of adenylyl cyclases mediated by distinct regulatory proteins. Nature. 1983 Apr 21;302(5910):706–709. doi: 10.1038/302706a0. [DOI] [PubMed] [Google Scholar]
  16. Hunter T., Sefton B. M. Transforming gene product of Rous sarcoma virus phosphorylates tyrosine. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1311–1315. doi: 10.1073/pnas.77.3.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jacobs S., Cuatrecasas P. Phosphorylation of receptors for insulin and insulin-like growth factor I. Effects of hormones and phorbol esters. J Biol Chem. 1986 Jan 15;261(2):934–939. [PubMed] [Google Scholar]
  18. Kadowaki T., Fujita-Yamaguchi Y., Nishida E., Takaku F., Akiyama T., Kathuria S., Akanuma Y., Kasuga M. Phosphorylation of tubulin and microtubule-associated proteins by the purified insulin receptor kinase. J Biol Chem. 1985 Apr 10;260(7):4016–4020. [PubMed] [Google Scholar]
  19. Kahn C. R., Baird K. L., Flier J. S., Grunfeld C., Harmon J. T., Harrison L. C., Karlsson F. A., Kasuga M., King G. L., Lang U. C. Insulin receptors, receptor antibodies, and the mechanism of insulin action. Recent Prog Horm Res. 1981;37:477–538. doi: 10.1016/b978-0-12-571137-1.50015-3. [DOI] [PubMed] [Google Scholar]
  20. Kapoor C. L., Chader G. J. Endogenous phosphorylation of retinal photoreceptor outer segment proteins by calcium phospholipid-dependent protein kinase. Biochem Biophys Res Commun. 1984 Aug 16;122(3):1397–1403. doi: 10.1016/0006-291x(84)91246-4. [DOI] [PubMed] [Google Scholar]
  21. Kasuga M., Karlsson F. A., Kahn C. R. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. doi: 10.1126/science.7031900. [DOI] [PubMed] [Google Scholar]
  22. Katada T., Gilman A. G., Watanabe Y., Bauer S., Jakobs K. H. Protein kinase C phosphorylates the inhibitory guanine-nucleotide-binding regulatory component and apparently suppresses its function in hormonal inhibition of adenylate cyclase. Eur J Biochem. 1985 Sep 2;151(2):431–437. doi: 10.1111/j.1432-1033.1985.tb09120.x. [DOI] [PubMed] [Google Scholar]
  23. Ku Y., Kishimoto A., Takai Y., Ogawa Y., Kimura S., Nishizuka Y. A new possible regulatory system for protein phosphorylation in human peripheral lymphocytes. II. Possible relation to phosphatidylinositol turnover induced by mitogens. J Immunol. 1981 Oct;127(4):1375–1379. [PubMed] [Google Scholar]
  24. Kühn H. Light- and GTP-regulated interaction of GTPase and other proteins with bovine photoreceptor membranes. Nature. 1980 Feb 7;283(5747):587–589. doi: 10.1038/283587a0. [DOI] [PubMed] [Google Scholar]
  25. Le Peuch C. J., Ballester R., Rosen O. M. Purified rat brain calcium- and phospholipid-dependent protein kinase phosphorylates ribosomal protein S6. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6858–6862. doi: 10.1073/pnas.80.22.6858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Manning D. R., Gilman A. G. The regulatory components of adenylate cyclase and transducin. A family of structurally homologous guanine nucleotide-binding proteins. J Biol Chem. 1983 Jun 10;258(11):7059–7063. [PubMed] [Google Scholar]
  27. Neer E. J., Lok J. M., Wolf L. G. Purification and properties of the inhibitory guanine nucleotide regulatory unit of brain adenylate cyclase. J Biol Chem. 1984 Nov 25;259(22):14222–14229. [PubMed] [Google Scholar]
  28. Northup J. K., Smigel M. D., Gilman A. G. The guanine nucleotide activating site of the regulatory component of adenylate cyclase. Identification by ligand binding. J Biol Chem. 1982 Oct 10;257(19):11416–11423. [PubMed] [Google Scholar]
  29. Ohkubo S., Yamada E., Endo T., Itoh H., Hidaka H. Vitamin A acid-induced activation of Ca2+-activated, phospholipid-dependent protein kinase from rabbit retina. Biochem Biophys Res Commun. 1984 Jan 30;118(2):460–466. doi: 10.1016/0006-291x(84)91325-1. [DOI] [PubMed] [Google Scholar]
  30. Petruzzelli L. M., Ganguly S., Smith C. J., Cobb M. H., Rubin C. S., Rosen O. M. Insulin activates a tyrosine-specific protein kinase in extracts of 3T3-L1 adipocytes and human placenta. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6792–6796. doi: 10.1073/pnas.79.22.6792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pines M., Gierschik P., Milligan G., Klee W., Spiegel A. Antibodies against the carboxyl-terminal 5-kDa peptide of the alpha subunit of transducin crossreact with the 40-kDa but not the 39-kDa guanine nucleotide binding protein from brain. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4095–4099. doi: 10.1073/pnas.82.12.4095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sagi-Eisenberg R., Pecht I. Protein kinase C, a coupling element between stimulus and secretion of basophils. Immunol Lett. 1984;8(5):237–241. doi: 10.1016/0165-2478(84)90002-6. [DOI] [PubMed] [Google Scholar]
  33. Spiegel A. M., Gierschik P., Levine M. A., Downs R. W., Jr Clinical implications of guanine nucleotide-binding proteins as receptor-effector couplers. N Engl J Med. 1985 Jan 3;312(1):26–33. doi: 10.1056/NEJM198501033120106. [DOI] [PubMed] [Google Scholar]
  34. Valentine-Braun K. A., Northup J. K., Hollenberg M. D. Epidermal growth factor (urogastrone)-mediated phosphorylation of a 35-kDa substrate in human placental membranes: relationship to the beta subunit of the guanine nucleotide regulatory complex. Proc Natl Acad Sci U S A. 1986 Jan;83(2):236–240. doi: 10.1073/pnas.83.2.236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Van Dop C., Yamanaka G., Steinberg F., Sekura R. D., Manclark C. R., Stryer L., Bourne H. R. ADP-ribosylation of transducin by pertussis toxin blocks the light-stimulated hydrolysis of GTP and cGMP in retinal photoreceptors. J Biol Chem. 1984 Jan 10;259(1):23–26. [PubMed] [Google Scholar]
  36. Yamazaki A., Stein P. J., Chernoff N., Bitensky M. W. Activation mechanism of rod outer segment cyclic GMP phosphodiesterase. Release of inhibitor by the GTP/GTP-binding protein. J Biol Chem. 1983 Jul 10;258(13):8188–8194. [PubMed] [Google Scholar]
  37. Zick Y., Kasuga M., Kahn C. R., Roth J. Characterization of insulin-mediated phosphorylation of the insulin receptor in a cell-free system. J Biol Chem. 1983 Jan 10;258(1):75–80. [PubMed] [Google Scholar]
  38. Zick Y., Whittaker J., Roth J. Insulin stimulated phosphorylation of its own receptor. Activation of a tyrosine-specific protein kinase that is tightly associated with the receptor. J Biol Chem. 1983 Mar 25;258(6):3431–3434. [PubMed] [Google Scholar]

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