The proliferative effects of gastrin on normal and neoplastic gastro-intestinal tissues have been shown to be mediated by the gastrin/CCKB (G/CCKB) G-protein-coupled receptors. We have recently reported that gastrin stimulates the tyrosine phosphorylation of Shc proteins and their subsequent association with the Grb2/Sos complex, leading to mitogen-activated protein kinase (MAPK) activation, a pathway known to play an important role in cell proliferation. We undertook the present study to characterize the signalling pathways used by this receptor to mediate the activation of the Shc/Grb2 complex. Since G/CCKB receptor occupancy leads to the activation of the phospholipase C (PLC)/protein kinase C (PKC) pathway, we examined whether PKC stimulation and Ca2+ mobilization contribute to the phosphorylation of Shc proteins and their association with Grb2 in response to gastrin. Our results indicate that Shc proteins are tyrosine phosphorylated and associate with Grb2 in response to phorbol esters, suggesting that activation of PKC is a potential signalling pathway leading to activation of the Shc/Grb2 complex. Inhibition of PKC by GF109203X completely blocked the effect of PMA on Shc tyrosine phosphorylation and its subsequent association with Grb2, but had a partial inhibitory effect on the response to gastrin. Depletion of the intracellular Ca2+ pools by treatment with thapsigargin blocked the increase in intracellular free calcium concentration induced by gastrin and diminished the ability of the peptide to stimulate Shc phosphorylation and recruitment of Grb2. In addition, removal of extracellular Ca2+ partially inhibited the effect of gastrin on Shc phosphorylation as well as its association with Grb2, indicating that the effects of gastrin are also mediated by Ca2+-dependent mechanisms. Furthermore, we show that blockage of the two major early signals generated by activation of PLC, which induced the activation of the Shc/Grb2 complex, also blocked gastrin-induced MAPK activation.
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Research Article|
July 15 1997
Ca2+ and protein kinase C-dependent mechanisms involved in gastrin-induced Shc/Grb2 complex formation and P44-mitogen-activated protein kinase activation
Laurence DAULHAC;
Laurence DAULHAC
1INSERM U.151, Groupe de Recherche de Biologie et Pathologie Digestive, Institut Louis Bugnard, CHU Rangueil, 31054 Toulouse, France
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Aline KOWALSKI-CHAUVEL;
Aline KOWALSKI-CHAUVEL
1INSERM U.151, Groupe de Recherche de Biologie et Pathologie Digestive, Institut Louis Bugnard, CHU Rangueil, 31054 Toulouse, France
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Lucien PRADAYROL;
Lucien PRADAYROL
1INSERM U.151, Groupe de Recherche de Biologie et Pathologie Digestive, Institut Louis Bugnard, CHU Rangueil, 31054 Toulouse, France
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Nicole VAYSSE;
Nicole VAYSSE
1INSERM U.151, Groupe de Recherche de Biologie et Pathologie Digestive, Institut Louis Bugnard, CHU Rangueil, 31054 Toulouse, France
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Catherine SEVA
Catherine SEVA
*
1INSERM U.151, Groupe de Recherche de Biologie et Pathologie Digestive, Institut Louis Bugnard, CHU Rangueil, 31054 Toulouse, France
*To whom correspondence should be addressed.
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Publisher: Portland Press Ltd
Received:
November 11 1996
Revision Received:
March 03 1997
Accepted:
March 04 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 325 (2): 383–389.
Article history
Received:
November 11 1996
Revision Received:
March 03 1997
Accepted:
March 04 1997
Citation
Laurence DAULHAC, Aline KOWALSKI-CHAUVEL, Lucien PRADAYROL, Nicole VAYSSE, Catherine SEVA; Ca2+ and protein kinase C-dependent mechanisms involved in gastrin-induced Shc/Grb2 complex formation and P44-mitogen-activated protein kinase activation. Biochem J 15 July 1997; 325 (2): 383–389. doi: https://doi.org/10.1042/bj3250383
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