Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Nov 15;320(Pt 1):87–91. doi: 10.1042/bj3200087

Bombesin and thrombin affect discrete pools of intracellular calcium through different G-proteins.

J L Wang 1, S Kalyanaraman 1, M D Vivo 1, N Gautam 1
PMCID: PMC1217901  PMID: 8947471

Abstract

In mouse NIH 3T3 cells, the mitogens bombesin and thrombin induced Ca2+ release from intracellular stores. Ca2+ release induced by bombesin was inhibited by the Ca(2+)-ATPase inhibitor thapsigargin, while Ca2+ release induced by thrombin was unaffected by this agent. The Ca(2+)-release response to bombesin was not affected by pertussis toxin, but the response to thrombin was abolished by the toxin. Stable transfectants overexpressing the G-protein subunit type alpha 9 showed an accentuated response to bombesin, indicating that the bombesin receptor was coupled to a Gq-like G-protein. Together, these results show that the two mitogenic receptors are coupled to distinct G-proteins that affect functionally different pools of Ca2+. Organization of signalling pathways in this manner may allow cells to differentially encode information from different signals.

Full Text

The Full Text of this article is available as a PDF (373.7 KB).

Selected References

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

  1. Balla T., Sim S. S., Iida T., Choi K. Y., Catt K. J., Rhee S. G. Agonist-induced calcium signaling is impaired in fibroblasts overproducing inositol 1,3,4,5-tetrakisphosphate. J Biol Chem. 1991 Dec 25;266(36):24719–24726. [PubMed] [Google Scholar]
  2. Battey J., Wada E. Two distinct receptor subtypes for mammalian bombesin-like peptides. Trends Neurosci. 1991 Dec;14(12):524–528. doi: 10.1016/0166-2236(91)90005-f. [DOI] [PubMed] [Google Scholar]
  3. Bian J. H., Ghosh T. K., Wang J. C., Gill D. L. Identification of intracellular calcium pools. Selective modification by thapsigargin. J Biol Chem. 1991 May 15;266(14):8801–8806. [PubMed] [Google Scholar]
  4. Cattaneo M. G., Magrini L., Sparber S. B., Vicentini L. M. Interaction between mitogens upon intracellular Ca2+ pools in murine fibroblasts. Cell Calcium. 1992 Nov;13(10):603–614. doi: 10.1016/0143-4160(92)90071-y. [DOI] [PubMed] [Google Scholar]
  5. Chambard J. C., Paris S., L'Allemain G., Pouysségur J. Two growth factor signalling pathways in fibroblasts distinguished by pertussis toxin. Nature. 1987 Apr 23;326(6115):800–803. doi: 10.1038/326800a0. [DOI] [PubMed] [Google Scholar]
  6. Charlesworth A., Rozengurt E. Thapsigargin and di-tert-butylhydroquinone induce synergistic stimulation of DNA synthesis with phorbol ester and bombesin in Swiss 3T3 cells. J Biol Chem. 1994 Dec 23;269(51):32528–32535. [PubMed] [Google Scholar]
  7. Clapham D. E. Calcium signaling. Cell. 1995 Jan 27;80(2):259–268. doi: 10.1016/0092-8674(95)90408-5. [DOI] [PubMed] [Google Scholar]
  8. Conklin B. R., Farfel Z., Lustig K. D., Julius D., Bourne H. R. Substitution of three amino acids switches receptor specificity of Gq alpha to that of Gi alpha. Nature. 1993 May 20;363(6426):274–276. doi: 10.1038/363274a0. [DOI] [PubMed] [Google Scholar]
  9. Corjay M. H., Dobrzanski D. J., Way J. M., Viallet J., Shapira H., Worland P., Sausville E. A., Battey J. F. Two distinct bombesin receptor subtypes are expressed and functional in human lung carcinoma cells. J Biol Chem. 1991 Oct 5;266(28):18771–18779. [PubMed] [Google Scholar]
  10. Coughlin S. R., Vu T. K., Hung D. T., Wheaton V. I. Characterization of a functional thrombin receptor. Issues and opportunities. J Clin Invest. 1992 Feb;89(2):351–355. doi: 10.1172/JCI115592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Vivo M., Chen J., Codina J., Iyengar R. Enhanced phospholipase C stimulation and transformation in NIH-3T3 cells expressing Q209LGq-alpha-subunits. J Biol Chem. 1992 Sep 15;267(26):18263–18266. [PubMed] [Google Scholar]
  12. Exton J. H. Phosphoinositide phospholipases and G proteins in hormone action. Annu Rev Physiol. 1994;56:349–369. doi: 10.1146/annurev.ph.56.030194.002025. [DOI] [PubMed] [Google Scholar]
  13. Gill D. L., Ghosh T. K., Bian J., Short A. D., Waldron R. T., Rybak S. L. Function and organization of the inositol 1,4,5-trisphosphate-sensitive calcium pool. Adv Second Messenger Phosphoprotein Res. 1992;26:265–308. [PubMed] [Google Scholar]
  14. Goligorsky M. S., Loftus D. J., Hruska K. A. Cytoplasmic calcium in individual proximal tubular cells in culture. Am J Physiol. 1986 Nov;251(5 Pt 2):F938–F944. doi: 10.1152/ajprenal.1986.251.5.F938. [DOI] [PubMed] [Google Scholar]
  15. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  16. Hamm H. E., Deretic D., Arendt A., Hargrave P. A., Koenig B., Hofmann K. P. Site of G protein binding to rhodopsin mapped with synthetic peptides from the alpha subunit. Science. 1988 Aug 12;241(4867):832–835. doi: 10.1126/science.3136547. [DOI] [PubMed] [Google Scholar]
  17. Harmon J. T., Jamieson G. A. Activation of platelets by alpha-thrombin is a receptor-mediated event. D-phenylalanyl-L-prolyl-L-arginine chloromethyl ketone-thrombin, but not N alpha-tosyl-L-lysine chloromethyl ketone-thrombin, binds to the high affinity thrombin receptor. J Biol Chem. 1986 Dec 5;261(34):15928–15933. [PubMed] [Google Scholar]
  18. Kasai H., Li Y. X., Miyashita Y. Subcellular distribution of Ca2+ release channels underlying Ca2+ waves and oscillations in exocrine pancreas. Cell. 1993 Aug 27;74(4):669–677. doi: 10.1016/0092-8674(93)90514-q. [DOI] [PubMed] [Google Scholar]
  19. Kisselev O., Gautam N. Specific interaction with rhodopsin is dependent on the gamma subunit type in a G protein. J Biol Chem. 1993 Nov 25;268(33):24519–24522. [PubMed] [Google Scholar]
  20. Kisselev O., Pronin A., Ermolaeva M., Gautam N. Receptor-G protein coupling is established by a potential conformational switch in the beta gamma complex. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9102–9106. doi: 10.1073/pnas.92.20.9102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lechleiter J., Girard S., Clapham D., Peralta E. Subcellular patterns of calcium release determined by G protein-specific residues of muscarinic receptors. Nature. 1991 Apr 11;350(6318):505–508. doi: 10.1038/350505a0. [DOI] [PubMed] [Google Scholar]
  22. Milligan G. Mechanisms of multifunctional signalling by G protein-linked receptors. Trends Pharmacol Sci. 1993 Jun;14(6):239–244. doi: 10.1016/0165-6147(93)90019-g. [DOI] [PubMed] [Google Scholar]
  23. Neer E. J. Heterotrimeric G proteins: organizers of transmembrane signals. Cell. 1995 Jan 27;80(2):249–257. doi: 10.1016/0092-8674(95)90407-7. [DOI] [PubMed] [Google Scholar]
  24. Petersen C. C., Toescu E. C., Petersen O. H. Different patterns of receptor-activated cytoplasmic Ca2+ oscillations in single pancreatic acinar cells: dependence on receptor type, agonist concentration and intracellular Ca2+ buffering. EMBO J. 1991 Mar;10(3):527–533. doi: 10.1002/j.1460-2075.1991.tb07979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Petersen O. H., Petersen C. C., Kasai H. Calcium and hormone action. Annu Rev Physiol. 1994;56:297–319. doi: 10.1146/annurev.ph.56.030194.001501. [DOI] [PubMed] [Google Scholar]
  26. Pronin A. N., Gautam N. Interaction between G-protein beta and gamma subunit types is selective. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6220–6224. doi: 10.1073/pnas.89.13.6220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Putney J. W., Jr Excitement about calcium signaling in inexcitable cells. Science. 1993 Oct 29;262(5134):676–678. doi: 10.1126/science.8235587. [DOI] [PubMed] [Google Scholar]
  28. Raben D. M., Yasuda K., Cunningham D. D. Modulation of thrombin-stimulated lipid responses in cultured fibroblasts. Evidence for two coupling mechanisms. Biochemistry. 1987 May 19;26(10):2759–2765. doi: 10.1021/bi00384a016. [DOI] [PubMed] [Google Scholar]
  29. Rhee S. G., Choi K. D. Regulation of inositol phospholipid-specific phospholipase C isozymes. J Biol Chem. 1992 Jun 25;267(18):12393–12396. [PubMed] [Google Scholar]
  30. Ross E. M. Signal sorting and amplification through G protein-coupled receptors. Neuron. 1989 Aug;3(2):141–152. doi: 10.1016/0896-6273(89)90027-5. [DOI] [PubMed] [Google Scholar]
  31. Schmidt C. J., Thomas T. C., Levine M. A., Neer E. J. Specificity of G protein beta and gamma subunit interactions. J Biol Chem. 1992 Jul 15;267(20):13807–13810. [PubMed] [Google Scholar]
  32. Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
  33. Sinnett-Smith J., Zachary I., Valverde A. M., Rozengurt E. Bombesin stimulation of p125 focal adhesion kinase tyrosine phosphorylation. Role of protein kinase C, Ca2+ mobilization, and the actin cytoskeleton. J Biol Chem. 1993 Jul 5;268(19):14261–14268. [PubMed] [Google Scholar]
  34. Smallridge R. C., Kiang J. G., Gist I. D., Fein H. G., Galloway R. J. U-73122, an aminosteroid phospholipase C antagonist, noncompetitively inhibits thyrotropin-releasing hormone effects in GH3 rat pituitary cells. Endocrinology. 1992 Oct;131(4):1883–1888. doi: 10.1210/endo.131.4.1396332. [DOI] [PubMed] [Google Scholar]
  35. Verma A., Hirsch D. J., Hanley M. R., Thastrup O., Christensen S. B., Snyder S. H. Inositol trisphosphate and thapsigargin discriminate endoplasmic reticulum stores of calcium in rat brain. Biochem Biophys Res Commun. 1990 Oct 30;172(2):811–816. doi: 10.1016/0006-291x(90)90747-b. [DOI] [PubMed] [Google Scholar]
  36. Vickers J. D. U73122 affects the equilibria between the phosphoinositides as well as phospholipase C activity in unstimulated and thrombin-stimulated human and rabbit platelets. J Pharmacol Exp Ther. 1993 Sep;266(3):1156–1163. [PubMed] [Google Scholar]
  37. Zachary I., Millar J., Nånberg E., Higgins T., Rozengurt E. Inhibition of bombesin-induced mitogenesis by pertussis toxin: dissociation from phospholipase C pathway. Biochem Biophys Res Commun. 1987 Jul 31;146(2):456–463. doi: 10.1016/0006-291x(87)90551-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES