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. 1994 Jul;94(1):328–336. doi: 10.1172/JCI117325

Regulation of hormone-sensitive calcium influx by the adenylyl cyclase system in renal epithelial cells.

K Kitamura 1, R T Miller 1
PMCID: PMC296313  PMID: 8040274

Abstract

To study signaling pathways regulated by alpha s and alpha i1 in renal epithelial cells, we expressed mutant, activated forms of alpha s and alpha i1 in a continuous proximal tubule cell line (MCT cells). alpha sQ227L increased cAMP production, and alpha ilQ204L reduced forskolin-sensitive cAMP production. alpha ilQ204L increased and alpha sQ227L decreased bradykinin-induced Ca influx across the cell membrane, but neither mutant affected bradykinin-stimulated intracellular Ca release or basal Ca influx. Bradykinin-stimulated Ca influx was reduced by dibutyryl cAMP, isoproterenol, and forskolin. Expression of a mutant regulatory type I subunit for cAMP-dependent protein kinase with reduced affinity for cAMP and treatment with KT-5720, a specific cAMP-dependent protein kinase inhibitor, enhanced Ca influx to a degree similar to that in cells expressing alpha ilQ204L. Bradykinin-stimulated c-fos mRNA expression is partially dependent on extracellular Ca. alpha sQ227L reduced and alpha ilQ204L enhanced bradykinin-stimulated c-fos expression. Consequently, in bradykinin-stimulated cells, the adenylyl cyclase system regulates Ca influx through cAMP-dependent protein kinase, but not intracellular Ca release. Furthermore, the Ca influx mechanism acts as an integrator of two signaling pathways such that Ca-dependent signals are damped by activators of adenylyl cyclase and enhanced by inhibitors of adenylyl cyclase.

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Selected References

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

  1. Aboolian A., Nord E. P. Bradykinin increases cytosolic free [Ca2+] in proximal tubule cells. Am J Physiol. 1988 Sep;255(3 Pt 2):F486–F493. doi: 10.1152/ajprenal.1988.255.3.F486. [DOI] [PubMed] [Google Scholar]
  2. Ashkenazi A., Winslow J. W., Peralta E. G., Peterson G. L., Schimerlik M. I., Capon D. J., Ramachandran J. An M2 muscarinic receptor subtype coupled to both adenylyl cyclase and phosphoinositide turnover. Science. 1987 Oct 30;238(4827):672–675. doi: 10.1126/science.2823384. [DOI] [PubMed] [Google Scholar]
  3. Bading H., Ginty D. D., Greenberg M. E. Regulation of gene expression in hippocampal neurons by distinct calcium signaling pathways. Science. 1993 Apr 9;260(5105):181–186. doi: 10.1126/science.8097060. [DOI] [PubMed] [Google Scholar]
  4. Bahnson T. D., Pandol S. J., Dionne V. E. Cyclic GMP modulates depletion-activated Ca2+ entry in pancreatic acinar cells. J Biol Chem. 1993 May 25;268(15):10808–10812. [PubMed] [Google Scholar]
  5. Berridge M. J. Cell signalling. A tale of two messengers. Nature. 1993 Sep 30;365(6445):388–389. doi: 10.1038/365388a0. [DOI] [PubMed] [Google Scholar]
  6. Berridge M. J. Inositol trisphosphate and calcium signalling. Nature. 1993 Jan 28;361(6410):315–325. doi: 10.1038/361315a0. [DOI] [PubMed] [Google Scholar]
  7. Birnbaumer L., Codina J., Mattera R., Yatani A., Scherer N., Toro M. J., Brown A. M. Signal transduction by G proteins. Kidney Int Suppl. 1987 Dec;23:S14–S42. [PubMed] [Google Scholar]
  8. Bourne H. R., Sanders D. A., McCormick F. The GTPase superfamily: a conserved switch for diverse cell functions. Nature. 1990 Nov 8;348(6297):125–132. doi: 10.1038/348125a0. [DOI] [PubMed] [Google Scholar]
  9. Brown A. M., Birnbaumer L. Direct G protein gating of ion channels. Am J Physiol. 1988 Mar;254(3 Pt 2):H401–H410. doi: 10.1152/ajpheart.1988.254.3.H401. [DOI] [PubMed] [Google Scholar]
  10. Carty D. J., Padrell E., Codina J., Birnbaumer L., Hildebrandt J. D., Iyengar R. Distinct guanine nucleotide binding and release properties of the three Gi proteins. J Biol Chem. 1990 Apr 15;265(11):6268–6273. [PubMed] [Google Scholar]
  11. Casey P. J., Fong H. K., Simon M. I., Gilman A. G. Gz, a guanine nucleotide-binding protein with unique biochemical properties. J Biol Chem. 1990 Feb 5;265(4):2383–2390. [PubMed] [Google Scholar]
  12. Clapham D. E. Cellular calcium. A mysterious new influx factor? Nature. 1993 Aug 26;364(6440):763–764. doi: 10.1038/364763a0. [DOI] [PubMed] [Google Scholar]
  13. Clegg C. H., Correll L. A., Cadd G. G., McKnight G. S. Inhibition of intracellular cAMP-dependent protein kinase using mutant genes of the regulatory type I subunit. J Biol Chem. 1987 Sep 25;262(27):13111–13119. [PubMed] [Google Scholar]
  14. Curran T., Gordon M. B., Rubino K. L., Sambucetti L. C. Isolation and characterization of the c-fos(rat) cDNA and analysis of post-translational modification in vitro. Oncogene. 1987;2(1):79–84. [PubMed] [Google Scholar]
  15. Federman A. D., Conklin B. R., Schrader K. A., Reed R. R., Bourne H. R. Hormonal stimulation of adenylyl cyclase through Gi-protein beta gamma subunits. Nature. 1992 Mar 12;356(6365):159–161. doi: 10.1038/356159a0. [DOI] [PubMed] [Google Scholar]
  16. Feinstein P. G., Schrader K. A., Bakalyar H. A., Tang W. J., Krupinski J., Gilman A. G., Reed R. R. Molecular cloning and characterization of a Ca2+/calmodulin-insensitive adenylyl cyclase from rat brain. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10173–10177. doi: 10.1073/pnas.88.22.10173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gao B. N., Gilman A. G. Cloning and expression of a widely distributed (type IV) adenylyl cyclase. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10178–10182. doi: 10.1073/pnas.88.22.10178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gil-Longo J., Dufour M. N., Guillon G., Lugnier C. G proteins in aortic endothelial cells and bradykinin-induced formation of nitric oxide. Eur J Pharmacol. 1993 Oct 15;247(2):119–125. doi: 10.1016/0922-4106(93)90068-k. [DOI] [PubMed] [Google Scholar]
  20. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  21. Graziano M. P., Gilman A. G. Synthesis in Escherichia coli of GTPase-deficient mutants of Gs alpha. J Biol Chem. 1989 Sep 15;264(26):15475–15482. [PubMed] [Google Scholar]
  22. 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]
  23. Haverty T. P., Kelly C. J., Hines W. H., Amenta P. S., Watanabe M., Harper R. A., Kefalides N. A., Neilson E. G. Characterization of a renal tubular epithelial cell line which secretes the autologous target antigen of autoimmune experimental interstitial nephritis. J Cell Biol. 1988 Oct;107(4):1359–1368. doi: 10.1083/jcb.107.4.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hermouet S., Merendino J. J., Jr, Gutkind J. S., Spiegel A. M. Activating and inactivating mutations of the alpha subunit of Gi2 protein have opposite effects on proliferation of NIH 3T3 cells. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10455–10459. doi: 10.1073/pnas.88.23.10455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jones D. T., Reed R. R. Molecular cloning of five GTP-binding protein cDNA species from rat olfactory neuroepithelium. J Biol Chem. 1987 Oct 15;262(29):14241–14249. [PubMed] [Google Scholar]
  26. Kase H., Iwahashi K., Nakanishi S., Matsuda Y., Yamada K., Takahashi M., Murakata C., Sato A., Kaneko M. K-252 compounds, novel and potent inhibitors of protein kinase C and cyclic nucleotide-dependent protein kinases. Biochem Biophys Res Commun. 1987 Jan 30;142(2):436–440. doi: 10.1016/0006-291x(87)90293-2. [DOI] [PubMed] [Google Scholar]
  27. Katada T., Northup J. K., Bokoch G. M., Ui M., Gilman A. G. The inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase. Subunit dissociation and guanine nucleotide-dependent hormonal inhibition. J Biol Chem. 1984 Mar 25;259(6):3578–3585. [PubMed] [Google Scholar]
  28. Katsushika S., Chen L., Kawabe J., Nilakantan R., Halnon N. J., Homcy C. J., Ishikawa Y. Cloning and characterization of a sixth adenylyl cyclase isoform: types V and VI constitute a subgroup within the mammalian adenylyl cyclase family. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8774–8778. doi: 10.1073/pnas.89.18.8774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  30. Linder M. E., Ewald D. A., Miller R. J., Gilman A. G. Purification and characterization of Go alpha and three types of Gi alpha after expression in Escherichia coli. J Biol Chem. 1990 May 15;265(14):8243–8251. [PubMed] [Google Scholar]
  31. Lowndes J. M., Gupta S. K., Osawa S., Johnson G. L. GTPase-deficient G alpha i2 oncogene gip2 inhibits adenylylcyclase and attenuates receptor-stimulated phospholipase A2 activity. J Biol Chem. 1991 Aug 5;266(22):14193–14197. [PubMed] [Google Scholar]
  32. Masters S. B., Miller R. T., Chi M. H., Chang F. H., Beiderman B., Lopez N. G., Bourne H. R. Mutations in the GTP-binding site of GS alpha alter stimulation of adenylyl cyclase. J Biol Chem. 1989 Sep 15;264(26):15467–15474. [PubMed] [Google Scholar]
  33. Muallem S., Khademazad M., Sachs G. The route of Ca2+ entry during reloading of the intracellular Ca2+ pool in pancreatic acini. J Biol Chem. 1990 Feb 5;265(4):2011–2016. [PubMed] [Google Scholar]
  34. Méry P. F., Brechler V., Pavoine C., Pecker F., Fischmeister R. Glucagon stimulates the cardiac Ca2+ current by activation of adenylyl cyclase and inhibition of phosphodiesterase. Nature. 1990 May 10;345(6271):158–161. doi: 10.1038/345158a0. [DOI] [PubMed] [Google Scholar]
  35. Neer E. J., Clapham D. E. Roles of G protein subunits in transmembrane signalling. Nature. 1988 May 12;333(6169):129–134. doi: 10.1038/333129a0. [DOI] [PubMed] [Google Scholar]
  36. Parekh A. B., Terlau H., Stühmer W. Depletion of InsP3 stores activates a Ca2+ and K+ current by means of a phosphatase and a diffusible messenger. Nature. 1993 Aug 26;364(6440):814–818. doi: 10.1038/364814a0. [DOI] [PubMed] [Google Scholar]
  37. Peralta E. G., Ashkenazi A., Winslow J. W., Ramachandran J., Capon D. J. Differential regulation of PI hydrolysis and adenylyl cyclase by muscarinic receptor subtypes. Nature. 1988 Aug 4;334(6181):434–437. doi: 10.1038/334434a0. [DOI] [PubMed] [Google Scholar]
  38. Premont R. T., Chen J., Ma H. W., Ponnapalli M., Iyengar R. Two members of a widely expressed subfamily of hormone-stimulated adenylyl cyclases. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9809–9813. doi: 10.1073/pnas.89.20.9809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Premont R. T., Jacobowitz O., Iyengar R. Lowered responsiveness of the catalyst of adenylyl cyclase to stimulation by GS in heterologous desensitization: a role for adenosine 3',5'-monophosphate-dependent phosphorylation. Endocrinology. 1992 Dec;131(6):2774–2784. doi: 10.1210/endo.131.6.1332848. [DOI] [PubMed] [Google Scholar]
  40. Randriamampita C., Tsien R. Y. Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature. 1993 Aug 26;364(6440):809–814. doi: 10.1038/364809a0. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Salomon Y., Londos C., Rodbell M. A highly sensitive adenylate cyclase assay. Anal Biochem. 1974 Apr;58(2):541–548. doi: 10.1016/0003-2697(74)90222-x. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
  45. Tang W. J., Gilman A. G. Adenylyl cyclases. Cell. 1992 Sep 18;70(6):869–872. doi: 10.1016/0092-8674(92)90236-6. [DOI] [PubMed] [Google Scholar]
  46. Tang W. J., Gilman A. G. Type-specific regulation of adenylyl cyclase by G protein beta gamma subunits. Science. 1991 Dec 6;254(5037):1500–1503. doi: 10.1126/science.1962211. [DOI] [PubMed] [Google Scholar]
  47. Trejo J., Brown J. H. c-fos and c-jun are induced by muscarinic receptor activation of protein kinase C but are differentially regulated by intracellular calcium. J Biol Chem. 1991 Apr 25;266(12):7876–7882. [PubMed] [Google Scholar]
  48. Tsien R. W., Tsien R. Y. Calcium channels, stores, and oscillations. Annu Rev Cell Biol. 1990;6:715–760. doi: 10.1146/annurev.cb.06.110190.003435. [DOI] [PubMed] [Google Scholar]
  49. Wong Y. H., Conklin B. R., Bourne H. R. Gz-mediated hormonal inhibition of cyclic AMP accumulation. Science. 1992 Jan 17;255(5042):339–342. doi: 10.1126/science.1347957. [DOI] [PubMed] [Google Scholar]
  50. Wong Y. H., Federman A., Pace A. M., Zachary I., Evans T., Pouysségur J., Bourne H. R. Mutant alpha subunits of Gi2 inhibit cyclic AMP accumulation. Nature. 1991 May 2;351(6321):63–65. doi: 10.1038/351063a0. [DOI] [PubMed] [Google Scholar]
  51. Yatani A., Codina J., Imoto Y., Reeves J. P., Birnbaumer L., Brown A. M. A G protein directly regulates mammalian cardiac calcium channels. Science. 1987 Nov 27;238(4831):1288–1292. doi: 10.1126/science.2446390. [DOI] [PubMed] [Google Scholar]

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