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. 1984 Jul;81(14):4316–4320. doi: 10.1073/pnas.81.14.4316

Phorbol ester induces desensitization of adenylate cyclase and phosphorylation of the beta-adrenergic receptor in turkey erythrocytes.

D J Kelleher, J E Pessin, A E Ruoho, G L Johnson
PMCID: PMC345579  PMID: 6087317

Abstract

Incubation of turkey erythrocytes with the phorbol ester phorbol 12-myristate 13-acetate (PMA) results in a dose- and time-dependent desensitization of isoproterenol-stimulated adenylate cyclase activity. Compared to controls, membranes from PMA-treated cells have an isoproterenol-stimulated adenylate cyclase activity that is decreased 20%-40%, with little effect on forskolin or fluoride activation of adenylate cyclase. No change in beta-adrenergic receptor number is observed after PMA treatment, indicating that the major effect of PMA is to uncouple receptor interactions with Ns, the stimulatory guanine nucleotide regulatory protein of adenylate cyclase. Purification of beta-adrenergic receptors from 32Pi-labeled turkey erythrocytes, incubated in the presence or absence of PMA, indicates that the phorbol ester is capable of inducing a 3-fold increase in phosphorylation of the beta-adrenergic receptor. The PMA effect is similar to the phosphorylation of the beta-adrenergic receptor during isoproterenol- and dibutyryl cAMP-induced desensitization of adenylate cyclase in turkey erythrocytes. The findings indicate that decreased receptor-Ns coupling is correlated with receptor phosphorylation and that phorbol esters can influence the responsiveness of hormone-sensitive adenylate cyclase in certain cell types.

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Selected References

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

  1. Alper S. L., Beam K. G., Greengard P. Hormonal control of Na+-K+ co-transport in turkey erythrocytes. Multiple site phosphorylation of goblin, a high molecular weight protein of the plasma membrane. J Biol Chem. 1980 May 25;255(10):4864–4871. [PubMed] [Google Scholar]
  2. Alper S. L., Palfrey H. C., DeRiemer S. A., Greengard P. Hormonal control of protein phosphorylation in turkey erythrocytes. Phosphorylation by cAMP-dependent and Ca2+-dependent protein kinases of distinct sites in goblin, a high molecular weight protein of the plasma membrane. J Biol Chem. 1980 Nov 25;255(22):11029–11039. [PubMed] [Google Scholar]
  3. Anderson W. B., Jaworski C. J. Isoproterenol-induced desensitization of adenylate cyclase responsiveness in a cell-free system. J Biol Chem. 1979 Jun 10;254(11):4596–4601. [PubMed] [Google Scholar]
  4. Blumberg P. M., Driedger P. E., Rossow P. W. Effect of a phorbol ester on a transformation-sensitive surface protein of chick fibroblasts. Nature. 1976 Dec 2;264(5585):446–447. doi: 10.1038/264446a0. [DOI] [PubMed] [Google Scholar]
  5. Blumberg P. M. In vitro studies on the mode of action of the phorbol esters, potent tumor promoters: part 1. Crit Rev Toxicol. 1980 Dec;8(2):153–197. doi: 10.3109/10408448009037493. [DOI] [PubMed] [Google Scholar]
  6. Bockaert J., Hunzicker-Dunn M., Birnbaumer L. Hormone-stimulated desensitization of hormone-dependent adenylyl cyclase. Dual action of luteninizing hormone on pig graafian follicle membranes. J Biol Chem. 1976 May 10;251(9):2653–2663. [PubMed] [Google Scholar]
  7. Brandt D. R., Asano T., Pedersen S. E., Ross E. M. Reconstitution of catecholamine-stimulated guanosinetriphosphatase activity. Biochemistry. 1983 Sep 13;22(19):4357–4362. doi: 10.1021/bi00288a002. [DOI] [PubMed] [Google Scholar]
  8. Brown K. D., Dicker P., Rozengurt E. Inhibition of epidermal growth factor binding to surface receptors by tumor promotors. Biochem Biophys Res Commun. 1979 Feb 28;86(4):1037–1043. doi: 10.1016/0006-291x(79)90221-3. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Cerione R. A., Strulovici B., Benovic J. L., Lefkowitz R. J., Caron M. G. Pure beta-adrenergic receptor: the single polypeptide confers catecholamine responsiveness to adenylate cyclase. Nature. 1983 Dec 8;306(5943):562–566. doi: 10.1038/306562a0. [DOI] [PubMed] [Google Scholar]
  11. Chandler C. E., Herschman H. R. Tumor promoter modulation of epidermal growth factor- and nerve growth factor-induced adhesion and growth factor binding of PC-12 pheochromocytoma cells. J Cell Physiol. 1980 Nov;105(2):275–285. doi: 10.1002/jcp.1041050211. [DOI] [PubMed] [Google Scholar]
  12. Diamond L., O'Brien S., Donaldson C., Shimizu Y. Growth stimulation of human diploid fibro-blasts by the tumor promoter, 12-0-tetradecanoly-phorbol-13-acetate. Int J Cancer. 1974 May 15;13(5):721–730. doi: 10.1002/ijc.2910130516. [DOI] [PubMed] [Google Scholar]
  13. End D., Tolson N., Yu M. Y., Guroff G. Effects of 12-0-Tetradecanoylphorbol-13-acetate (TPA) on rat pheochromocytoma (PC12) cells: interactions with epidermal growth factor and nerve growth factor. J Cell Physiol. 1982 May;111(2):140–148. doi: 10.1002/jcp.1041110204. [DOI] [PubMed] [Google Scholar]
  14. Green D. A., Clark R. B. Adenylate cyclase coupling proteins are not essential for agonist-specific desensitization of lymphoma cells. J Biol Chem. 1981 Mar 10;256(5):2105–2108. [PubMed] [Google Scholar]
  15. Grunberger G., Gorden P. Affinity alteration of insulin receptor induced by a phorbol ester. Am J Physiol. 1982 Oct;243(4):E319–E324. doi: 10.1152/ajpendo.1982.243.4.E319. [DOI] [PubMed] [Google Scholar]
  16. Hoffman B. B., Mullikin-Kilpatrick D., Lefkowitz R. J. Desensitization of beta-adrenergic stimulated adenylate cyclase in turkey erythrocytes. J Cyclic Nucleotide Res. 1979 Oct;5(5):355–366. [PubMed] [Google Scholar]
  17. Huberman E., Weeks C., Herrmann A., Callaham M., Slaga T. Alterations in polyamine levels induced by phorbol diesters and other agents that promote differentiation in human promyelocytic leukemia cells. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1062–1066. doi: 10.1073/pnas.78.2.1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hudson T. H., Johnson G. L. Functional alterations in components of pigeon erythrocyte adenylate cyclase following desensitization to isoproterenol. Mol Pharmacol. 1981 Nov;20(3):694–703. [PubMed] [Google Scholar]
  19. Iyengar R., Mintz P. W., Swartz T. L., Birnbaumer L. Divalent cation-induced desensitization of glucagon-stimulable adenylyl cyclase in rat liver plasma membrane. GTP-dependent stimulation by glucagon. J Biol Chem. 1980 Dec 25;255(24):11875–11882. [PubMed] [Google Scholar]
  20. Jacobs S., Sahyoun N. E., Saltiel A. R., Cuatrecasas P. Phorbol esters stimulate the phosphorylation of receptors for insulin and somatomedin C. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6211–6213. doi: 10.1073/pnas.80.20.6211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kelleher D. J., Rashidbaigi A., Ruoho A. E., Johnson G. L. Rapid vesicle reconstitution of alprenolol-Sepharose-purified beta 1-adrenergic receptors. Interaction of the purified receptor with N. J Biol Chem. 1983 Nov 10;258(21):12881–12885. [PubMed] [Google Scholar]
  22. King A. C., Cuatrecasas P. Resolution of high and low affinity epidermal growth factor receptors. Inhibition of high affinity component by low temperature, cycloheximide, and phorbol esters. J Biol Chem. 1982 Mar 25;257(6):3053–3060. [PubMed] [Google Scholar]
  23. Koeffler H. P., Bar-Eli M., Territo M. Phorbol diester-induced macrophage differentiation of leukemic blasts from patients with human myelogenous leukemia. J Clin Invest. 1980 Nov;66(5):1101–1108. doi: 10.1172/JCI109939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kraft A. S., Anderson W. B., Cooper H. L., Sando J. J. Decrease in cytosolic calcium/phospholipid-dependent protein kinase activity following phorbol ester treatment of EL4 thymoma cells. J Biol Chem. 1982 Nov 25;257(22):13193–13196. [PubMed] [Google Scholar]
  25. Lee L. S., Weinstein I. B. Mechanism of tumor promoter inhibition of cellular binding of epidermal growth factor. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5168–5172. doi: 10.1073/pnas.76.10.5168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lefkowitz R. J., Stadel J. M., Caron M. G. Adenylate cyclase-coupled beta-adrenergic receptors: structure and mechanisms of activation and desensitization. Annu Rev Biochem. 1983;52:159–186. doi: 10.1146/annurev.bi.52.070183.001111. [DOI] [PubMed] [Google Scholar]
  27. Lotem J., Sachs L. Regulation of normal differentiation in mouse and human myeloid leukemic cells by phorbol esters and the mechanism of tumor promotion. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5158–5162. doi: 10.1073/pnas.76.10.5158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mallorga P., Tallman J. F., Henneberry R. C., Hirata F., Strittmatter W. T., Axelrod J. Mepacrine blocks beta-adrenergic agonist-induced desensitization in astrocytoma cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1341–1345. doi: 10.1073/pnas.77.3.1341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mufson R. A., Simsiman R. C., Boutwell R. K. The effect of the phorbol ester tumor promoters on the basal and catecholamine-stimulated levels of cyclic adenosine 3':5'-monophosphate in mouse skin and epidermis in vivo. Cancer Res. 1977 Mar;37(3):665–669. [PubMed] [Google Scholar]
  30. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pegoraro L., Abrahm J., Cooper R. A., Levis A., Lange B., Meo P., Rovera G. Differentiation of human leukemias in response to 12-0-tetradecanoylphorbol-13-acetate in vitro. Blood. 1980 May;55(5):859–862. [PubMed] [Google Scholar]
  32. Ross E. M. Phosphatidylcholine-promoted interaction of the catalytic and regulatory proteins of adenylate cyclase. J Biol Chem. 1982 Sep 25;257(18):10751–10758. [PubMed] [Google Scholar]
  33. Roy C., Guillon G., Jard S. Hormone-dependent desensitization of vasopressin-sensitive adenylate cyclase. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1265–1270. doi: 10.1016/s0006-291x(76)80151-9. [DOI] [PubMed] [Google Scholar]
  34. Shear M., Insel P. A., Melmon K. L., Coffino P. Agonist-specific refractoriness induced by isoproterenol. Studies with mutant cells. J Biol Chem. 1976 Dec 10;251(23):7572–7576. [PubMed] [Google Scholar]
  35. Shorr R. G., Strohsacker M. W., Lavin T. N., Lefkowitz R. J., Caron M. G. The beta 1-adrenergic receptor of the turkey erythrocyte. Molecular heterogeneity revealed by purification and photoaffinity labeling. J Biol Chem. 1982 Oct 25;257(20):12341–12350. [PubMed] [Google Scholar]
  36. Shoyab M., De Larco J. E., Todaro G. J. Biologically active phorbol esters specifically alter affinity of epidermal growth factor membrane receptors. Nature. 1979 May 31;279(5712):387–391. doi: 10.1038/279387a0. [DOI] [PubMed] [Google Scholar]
  37. Simpson I. A., Pfeuffer T. Functional desensitisation of beta-adrenergic receptors of avian erythrocytes by catecholamines and adenosine 3',5'-phosphate. Eur J Biochem. 1980 Oct;111(1):111–116. doi: 10.1111/j.1432-1033.1980.tb06081.x. [DOI] [PubMed] [Google Scholar]
  38. Stadel J. M., Nambi P., Shorr R. G., Sawyer D. F., Caron M. G., Lefkowitz R. J. Catecholamine-induced desensitization of turkey erythrocyte adenylate cyclase is associated with phosphorylation of the beta-adrenergic receptor. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3173–3177. doi: 10.1073/pnas.80.11.3173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Strulovici B., Stadel J. M., Lefkowitz R. J. Functional integrity of desensitized beta-adrenergic receptors. J Biol Chem. 1983 May 25;258(10):6410–6414. [PubMed] [Google Scholar]
  40. Su Y. F., Harden T. K., Perkins J. P. Isoproterenol-induced desensitization of adenylate cyclase in human astrocytoma cells. Relation of loss of hormonal responsiveness and decrement in beta-adrenergic receptors. J Biol Chem. 1979 Jan 10;254(1):38–41. [PubMed] [Google Scholar]
  41. Wertz P. W., Mueller G. C. Rapid stimulation of phospholipid metabolism in bovine lymphocytes by tumor-promoting phorbol esters. Cancer Res. 1978 Sep;38(9):2900–2904. [PubMed] [Google Scholar]
  42. Yuspa S. H., Lichti U., Ben T., Patterson E., Hennings H., Slaga T. J., Colburn N., Kelsey W. Phorbol esters stimulate DNA synthesis and ornithine decarboxylase activity in mouse epidermal cell cultures. Nature. 1976 Jul 29;262(5567):402–404. doi: 10.1038/262402a0. [DOI] [PubMed] [Google Scholar]

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