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Regulation of CCK-induced ERK1/2 activation by PKC epsilon in rat pancreatic acinar cells

Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109-0622, USA

The extracellular signal-regulated kinase ERK1/2 is activated in pancreatic acinar cells by cholecystokinin (CCK) and other secretagogues with this activation mediated primarily by protein kinase C (PKC). To identify the responsible PKC isoform, we utilized chemical inhibitors, cell permeant inhibitory peptides and overexpression of individual PKC dominant negative variants by means of adenoviral vectors. While the broad-spectrum PKC inhibitor GF109203X strongly inhibited ERK1/2 activation induced by 100 pM CCK, Go6976 which inhibits the classical PKC isoforms (alpha, beta and gamma), as well as Rottlerin, a specific PKC delta inhibitor, had no inhibitory effect. To test the role of PKC epsilon, we used specific cell permeant peptide inhibitors which block PKC interaction with their intracellular receptors or RACKs. Only PP93 (PKC epsilon peptide inhibitor) inhibited CCK-induced ERK1/2 activation, while PP95, PP101 and PP98, which are PKC alpha, delta and zeta peptide inhibitors respectively, had no effect. We also utilized adenovirus to express dominant negative PKC isoforms in pancreatic acini. Only PKC epsilon dominant negative inhibited CCK-induced ERK1/2 activation. Dominant negative PKC epsilon expression similarly blocked the effect of carbachol and bombesin to activate ERK1/2. Immunoprecipitation results demonstrated that CCK can induce an interaction of c-Raf-1 and PKC epsilon, but not that of other isoforms of Raf or PKC. We conclude that PKC epsilon is the isoform of PKC primarily involved with CCK-induced ERK1/2 activation in pancreatic acinar cells.
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Keywords cholecystokinin; carbachol; protein kinase C; ERK1/2; adenoviral vectors

Citation: Chenwei Li, John A. Williams. Regulation of CCK-induced ERK1/2 activation by PKC epsilon in rat pancreatic acinar cells. AIMS Molecular Science, 2017, 4(4): 463-477. doi: 10.3934/molsci.2017.4.463

References

  • 1. Roskoski R Jr (2012) ERK1/2 MAP kinases: Structure, function, and regulation. Pharmacol Res 66: 105-143.    
  • 2. Krishna M, Narang H (2008) The complexity of mitogen-activated protein kinases (MAPKs) made simple. Cell Mol Life Sci 65: 3525-3544.    
  • 3. Yao Z, Seger R (2009) The ERK signaling cascade–views from different subcellular compartments. Biofactors 35: 407-416.    
  • 4. Duan RD, Williams JA (1994) Cholecystokinin rapidly activates mitogen-activated protein kinase in rat pancreatic acini. Am J Physiol 267: 401-408.
  • 5. Dabrowski A, Groblewski GE, Schafer C, et al. (1997) Cholecystokinin and EGF activate a MAPK cascade by different mechanisms in rat pancreatic acinar cells. Am J Physiol 273: C1472-C1479.
  • 6. Daulhac L, Kowalski-Chauvel A, Pradayrol L, et al. (1997) Ca2+ and protein kinase C-dependent mechanisms involved in gastrin-induced Shc/Grb2 complex formation and P44-mitogen-activated protein kinase activation. Biochem J 325: 383-389.    
  • 7. Piiper A, Gebhardt R, Kronenberger B, et al. (2000) Pertussis Toxin Inhibits Cholecytoskinin- and Epidermal Growth Factor-Induced Mitogen-Activated Protein Kinase Activation by Disinhibitation of the cAMP Signaling Pathway and Inhibition of c-Raf-1. Mol Pharmacol 58: 608-613.
  • 8. Piiper A, Elez R, You S, et al. (2003) Cholecystokinin Stimulates Extracellular Signal-regulated Kinase through Activation of the Epidermal Growth Factor Receptor, Yes, and Protein Kinase C. J Biol Chem 278: 7065-7072.    
  • 9. Koh Y, Tamizhselvi R, Bhatia M (2009) Extracellular Signal-Regulated Kinase 1/2 and c-Jun NH2-Terminal Kinase, through Nuclear Factor-kB and Activator Protein-1, Contribute to Caerulein-Induced Expression of Substance P and Neurokinin-1 Receptors in Pancreatic Acinar Cells. JPET 332: 940-948.
  • 10. Williams JA (2008) Receptor-mediated signal transduction pathways and the regulation of pancreatic acinar cell function. Curr Opin Gastroenterol 24: 573-579.    
  • 11. Jacob C, Bunnet NW (2006) Transmembrane Signaling by G Protein-Coupled Receptors, In: Physiology of the Gastrointestinal Tract, 4 Eds., Academic Press, 63-85.
  • 12. Williams JA, Yule DI (2012) Stimulus-secretion Coupling in Pancreatic Acinar Cells, In: Physiology of the Gastrointestinal Tract, 5 Eds., Elsevier, 1361-1398.
  • 13. Ramos JW (2008) The regulation of extracellular signal-regulated kinase (ERK) in mammalian cells. Int J Biochem Cell Biol 40: 2707-2719.    
  • 14. Nicke B, Tseng MJ, Fenrich M, et al. (1999) Adenovirus-mediated gene transfer of RasN17 inhibits specific CCK actions on pancreatic acinar cells. Am Physiol 276: 499-506.
  • 15. Nishizuka Y (1992) Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258: 607-614.    
  • 16. Newton AC (2009) Protein kinase C: poised to signal. Am J Physiol Endocrinol Metab 298: E395-E402.
  • 17. Mochly-Rosen D (1995) Localization of protein kinases by anchoring proteins: a theme in signal transduction. Science 268: 247-251.    
  • 18. Mochly-Rosen D, Wu G, Hahn H, et al. (2000) Cardiotrophic effects of protein kinase C-analysis in vivo modulation of PKCε Translocation. Circ Res 86: 1172-1179.
  • 19. Wu-Zhang AX, Newton AC (2013) Protein kinase C pharmacology: refining the toolbox. Biochem J 452: 195-209.    
  • 20. Churchill EN, Qvit N, Mochly-Rosen D (2009) Rationally designed peptide regulators of protein kinase C. Trends Endocrinol Metab 20: 25-33.    
  • 21. Bastani B, Yang L, Baldassare JJ, et al. (1995) Cellular distribution of isoforms of protein kinase C (PCK) in pancreatic acini. Biochimica et Biophysica Acta 1269: 307-315.    
  • 22. Kim MJ, Lee YS, Lee KH, et al. (2001) Site-specific localization of protein kinase C isoforms in rat pancreas. Pancreatology 1: 36-42    
  • 23. Li C, Chen X, Williams JA (2004) Regulation of CCK-induced amylase release by PKC-δ in rat pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol 287: G764-G771.    
  • 24. Rodriguez-Martin E, Boyano-Adanex MC, Bodega G, et al. (1999) Redistribution of protein kinase C isoforms in rat pancreatic acini during lactation and weaning. FEBS Lett 445: 356-360.    
  • 25. Fleming AK, Storz P (2017) Protein kinase C isoforms in the normal pancreas and in pancreatic disease. Cell Signal 40: 1-9    
  • 26. Braz JC, Bueno OF, De Windt LJ, et al. (2002) PKC alpha regulates the hypertrophic growth of cardiomyocytes through extracellular signal-regulated kinase1/2 (ERK1/2). J Cell Biol 156: 905-919.    
  • 27. Chen X, Edwards JA, Logsdon CD, et al. (2002) Dominant negative Rab3D inhibits amylase release from mous pancreatic acini. J Biol Chem 277: 18002-18009.    
  • 28. Besson A, Davy A, Robbins SM, et al. (2001) Differential activation of ERKs to focal adhesions by PKC epsilon is required for PMA-induced adhesion and migration of human glioma cells. Oncogene 20: 7398-7407.    
  • 29. Ginnan R, Pfleiderer PJ, Pumiglia K, et al. (2004) PKC-delta and CaMKII-delta 2 mediate ATP-dependent activation of ERK1/2 in vascular smooth muscle. Am J Physiol Cell Physiol 286: C1281-C1289.    
  • 30. Kampfer S, Windegger M, Hochholdinger F, et al. (2001) Protein kinase C isoforms involved in the transcriptional activation of cyclin D1 by transforming Ha-Ras. J Biol Chem 276: 42834-42842.    
  • 31. Torricelli C, Valacchi G, Maioli E (2001) Novel PKCs activate ERK through PKD1 in MCF-7 cells. In Vitro Cell Dev Bio-Animal 47: 73-81.
  • 32. Olson ER, Shamhart PE, Naugle JE, et al. (2008) Angiotensin II-induced extracellular signal-regulated kinase 1/2 activation is mediated by protein kinase Cdelta and intracellular calcium in adult rat cardiac fibroblasts. Hypertension 51: 704-711.    
  • 33. Gschwendt M, Muller HJ, Kielbassa K, et al. (1994) Rottlerin, a novel protein kinase inhibitor. Biochem Biophys Res Commun 199: 93-98.    
  • 34. Martiny-Baron G, Kazanietz MG, Mischak H, et al. (1993) Selective inhibition of protein kinase C isozymes by the indolocarbazol Go 6976. J Biol Chem 268: 9194-9197.
  • 35. Tapia JA, Jensen RT, Garcia-Marin LJ (2006) Rottlerin inhibits stimulated enzymatic secretion and several intracellular signaling transduction pathways in pancreatic acinar cells by a non-PKC-delta-dependent mechanism. Biochim Biophys Acta 1763: 25-38.    
  • 36. Chen L, Hahn H, Wu G, et al. (2001) Opposing cardioprotective actions and parallel hypertrophic effects of δPKC and εPKC. PNAS 98: 11114-11119.    
  • 37. Churchill E, Budas G, Vallentin A, et al. (2008) PKC isozymes in chronic cardiac disease: possible therapeutic targets. Annu Rev Pharmacol Toxicol 48: 569-599.    
  • 38. Chen L, Mochly-Rosen D (2001) Opposing effects δ of ξ and PKC in ethanol-induced cardioprotection. J Mol Cell Cardiol 33: 581-585.    
  • 39. Budas GR, Churchill EN, Mochly-Rosen D (2007) Cardioprotective mechanisms of PKC isozyme-selective activators and inhibitors in the treatment of ischemia-reperfusion injury. Pharm Res 55: 523-536.    
  • 40. Sabbatini ME, Chen X, Ernst SA, et al. (2008) Rap1 activation plays a regulatory role in pancreatic amylase secretion. J Biol Chem 283: 23884-23894.    
  • 41. Corbit KC, Trakul N, Eves EM, et al. (2003) Activation of Raf-1 signaling by protein kinase C through a mechanism involving Raf Kinase Inhibitory Protein. J Biol Chem 278: 13061-13068.    
  • 42. Han B, Ji B, Logsdon CD (2001) CCK independently activates intracellular trypsinogen and NF-kappaB in rat pancreatic acinar cells. Am J Physiol Cell Physiol 180: C465-C472.
  • 43. Satoh A, Gukovskaya AS, Nieto JM, et al. (2004) PKC-delta and -epsilon regulate NF-kappaB activation induced by cholecystokinin and TNF-alpha in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol 387: G582-G591.
  • 44. Thrower EC, Osgood S, Shugrue CA, et al. (2008) The novel protein kinase C isoforms -delta and -epsilon modulate caerulein-induced zymogen activation in pancreatic acinar cells. Am J Physicol Gastrointest Liver Physiol 294: G1344-1353.    
  • 45. Uchida T, Iwashita N, Ohara-Imaizumi M, et al. (2007) Protein kinase Cδ plays a non-redundant role in insulin secretion in pancreatic beta cells. J Biol Chem 282: 2707-2716.    
  • 46. Thrower EC, Wang J, Cheriyan S, et al. (2009) Protein kinase C δ-mediated processes in cholecystokinin-8-stimulated pancreatic acini. Pancreas 38: 930-935.    
  • 47. Cosen-Binker LI, Lam PP, Binker MG, et al. (2007) Alcohol/cholecystokinin-evoked pancreatic acinar basolateral exocytosis is mediated by protein kinase C alpha phosphorylation of Munc18c. J Biol Chem 282: 13047-13058.    

 

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