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. 1994 Dec 1;304(Pt 2):611–616. doi: 10.1042/bj3040611

Protein kinase C-dependent cyclic AMP formation in airway smooth muscle: the role of type II adenylate cyclase and the blockade of extracellular-signal-regulated kinase-2 (ERK-2) activation.

N J Pyne 1, N Moughal 1, P A Stevens 1, D Tolan 1, S Pyne 1
PMCID: PMC1137535  PMID: 7998998

Abstract

Bradykinin activates adenylate cyclase via a pathway that involves the 'up-stream' regulation of phospholipase D (PLD)-catalysed hydrolysis of phosphatidylcholine and activation of protein kinase C (PKC) in airway smooth muscle [Stevens, Pyne, Grady and Pyne (1994) Biochem. J. 297, 233-239]. Coincident signal (Gs alpha and PKC) amplification of the cyclic AMP response can be completely attenuated either by diverting PLD-derived phosphatidate or by inhibiting PKC. In this regard, the coincident signal detector type II adenylate cyclase is expressed as a 110/112 kDa polypeptide in these cells. PKC alpha is not involved in the activation of adenylate cyclase, since a B2-receptor antagonist (NPC567, 10 microM) blocked its bradykinin-stimulated translocation to the membrane and was without effect against both bradykinin-stimulated PLD activity and cyclic AMP formation. Cyclic AMP formation can also be activated by platelet-derived growth factor (PDGF), via a PKC-dependent pathway, although the magnitude of the response is less than that elicited by bradykinin. Nevertheless, these results indicate that multiple receptor types employ PKC to initiate cyclic AMP signals. PDGF (10 ng/ml) elicited the marked sustained activation of extracellular-signal-regulated kinase-2 (ERK-2), whereas bradykinin (1 microM) provoked only modest transient activation of ERK-2. Deoxyadenosine (0.1 mM), a P-site inhibitor of adenylate cyclase, blocked bradykinin-stimulated cyclic AMP formation and converted the activation of ERK-2 into a sustained response. Thus the PKC-stimulated cyclic AMP response can limit the activation of ERK-2 in response to bradykinin. These studies indicate that the integration of distinct signal pathways by adenylate cyclase can determine the kinetics of ERK activation, an enzyme that appears to be important for mitogenic progression.

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Selected References

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

  1. Boulton T. G., Cobb M. H. Identification of multiple extracellular signal-regulated kinases (ERKs) with antipeptide antibodies. Cell Regul. 1991 May;2(5):357–371. doi: 10.1091/mbc.2.5.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen J., Iyengar R. Inhibition of cloned adenylyl cyclases by mutant-activated Gi-alpha and specific suppression of type 2 adenylyl cyclase inhibition by phorbol ester treatment. J Biol Chem. 1993 Jun 15;268(17):12253–12256. [PubMed] [Google Scholar]
  3. Cook S. J., McCormick F. Inhibition by cAMP of Ras-dependent activation of Raf. Science. 1993 Nov 12;262(5136):1069–1072. doi: 10.1126/science.7694367. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Grady M., Stevens P. A., Pyne S., Pyne N. Adenylyl cyclase in lung from hypersensitive guinea pig displays increased responsiveness to guanine nucleotides and isoprenaline: the role of the G proteins Gs and Gi. Biochim Biophys Acta. 1993 Apr 16;1176(3):313–320. doi: 10.1016/0167-4889(93)90060-3. [DOI] [PubMed] [Google Scholar]
  7. Howe L. R., Marshall C. J. Lysophosphatidic acid stimulates mitogen-activated protein kinase activation via a G-protein-coupled pathway requiring p21ras and p74raf-1. J Biol Chem. 1993 Oct 5;268(28):20717–20720. [PubMed] [Google Scholar]
  8. Iyengar R. Molecular and functional diversity of mammalian Gs-stimulated adenylyl cyclases. FASEB J. 1993 Jun;7(9):768–775. doi: 10.1096/fasebj.7.9.8330684. [DOI] [PubMed] [Google Scholar]
  9. Jacobowitz O., Chen J., Premont R. T., Iyengar R. Stimulation of specific types of Gs-stimulated adenylyl cyclases by phorbol ester treatment. J Biol Chem. 1993 Feb 25;268(6):3829–3832. [PubMed] [Google Scholar]
  10. Kolch W., Heidecker G., Kochs G., Hummel R., Vahidi H., Mischak H., Finkenzeller G., Marmé D., Rapp U. R. Protein kinase C alpha activates RAF-1 by direct phosphorylation. Nature. 1993 Jul 15;364(6434):249–252. doi: 10.1038/364249a0. [DOI] [PubMed] [Google Scholar]
  11. Lustig K. D., Conklin B. R., Herzmark P., Taussig R., Bourne H. R. Type II adenylylcyclase integrates coincident signals from Gs, Gi, and Gq. J Biol Chem. 1993 Jul 5;268(19):13900–13905. [PubMed] [Google Scholar]
  12. Pyne S., Pyne N. J. Bradykinin stimulates phospholipase D in primary cultures of guinea-pig tracheal smooth muscle. Biochem Pharmacol. 1993 Feb 9;45(3):593–603. doi: 10.1016/0006-2952(93)90132-g. [DOI] [PubMed] [Google Scholar]
  13. Pyne S., Pyne N. J. Bradykinin-stimulated phosphatidate and 1,2-diacylglycerol accumulation in guinea-pig airway smooth muscle: evidence for regulation 'down-stream' of phospholipases. Cell Signal. 1994 Mar;6(3):269–277. doi: 10.1016/0898-6568(94)90031-0. [DOI] [PubMed] [Google Scholar]
  14. Pyne S., Pyne N. J. Differential effects of B2 receptor antagonists upon bradykinin-stimulated phospholipase C and D in guinea-pig cultured tracheal smooth muscle. Br J Pharmacol. 1993 Sep;110(1):477–481. doi: 10.1111/j.1476-5381.1993.tb13835.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rubin C. S., Erlichman J., Rosen O. M. Cyclic AMP-dependent protein kinase from bovine heart muscle. Methods Enzymol. 1974;38:308–315. doi: 10.1016/0076-6879(74)38047-0. [DOI] [PubMed] [Google Scholar]
  16. Stevens P. A., Pyne S., Grady M., Pyne N. J. Bradykinin-dependent activation of adenylate cyclase activity and cyclic AMP accumulation in tracheal smooth muscle occurs via protein kinase C-dependent and -independent pathways. Biochem J. 1994 Jan 1;297(Pt 1):233–239. doi: 10.1042/bj2970233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wu J., Dent P., Jelinek T., Wolfman A., Weber M. J., Sturgill T. W. Inhibition of the EGF-activated MAP kinase signaling pathway by adenosine 3',5'-monophosphate. Science. 1993 Nov 12;262(5136):1065–1069. doi: 10.1126/science.7694366. [DOI] [PubMed] [Google Scholar]
  18. Xia Z., Choi E. J., Wang F., Blazynski C., Storm D. R. Type I calmodulin-sensitive adenylyl cyclase is neural specific. J Neurochem. 1993 Jan;60(1):305–311. doi: 10.1111/j.1471-4159.1993.tb05852.x. [DOI] [PubMed] [Google Scholar]

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