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J. Biol. Chem., Vol. 277, Issue 25, 22407-22413, June 21, 2002
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From INSERM Unité 45 and IFR 62, Hôpital Edouard
Herriot, Pavillon H, F-69437 Lyon Cedex 3, France
Little is known about the mechanisms by which
protein-derived nutrients regulate hormone gene expression in the
intestine. We have previously reported that protein hydrolysates
(i.e. peptones), which are representative of the protein
fraction in the lumen, increased cholecystokinin
(CCK) gene transcription in the STC-1 enteroendocrine cell
line. In the present work, we examined the intracellular events evoked
by peptones to stimulate CCK gene transcription. In STC-1
cells, peptones stimulated cyclic AMP production and protein kinase A
(PKA) activity. This was associated with a nuclear translocation of the
PKA catalytic subunit and with a PKA-dependent
phosphorylation of the CRE-binding protein (CREB) at
Ser133. Using transient transfection experiments and
reporter luciferase assays, we show that peptone-stimulated
transcriptional activity of the CCK gene promoter was
significantly decreased when the PKA pathway was inhibited.
Furthermore, the intracellular calcium chelator
1,2-bis-(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-tetra(acetoxymethyl)ester completely inhibited peptone-induced stimulation of the CCK gene promoter activity,
phosphorylation of CREB, and PKA activity. Peptones increased, in a
calcium-dependent manner, the phosphorylation of
extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and the
MEK inhibitor PD98059 decreased the peptone-induced stimulation of
CCK gene promoter activity. This stimulation was also
reduced by 30% in the presence of the
calcium/calmodulin-dependent protein kinase (CaMK)
inhibitor KN-93. Total inhibition was obtained when the PKA, ERK, and
CaMK pathways were simultaneously blocked with appropriate inhibitors to these pathways. These results demonstrate the simultaneous involvement of cAMP- and calcium-dependent protein kinases
in the stimulation of intestinal CCK gene transcription by
protein-derived nutrients.
Co-requirement of Cyclic AMP- and Calcium-dependent
Protein Kinases for Transcriptional Activation of Cholecystokinin Gene
by Protein Hydrolysates*
,
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a Ph.D. grant from the French Ministère de
l'Education Nationale de la Recherche et de la Technologie.
§
To whom correspondence should be addressed: INSERM Unité 45, Hôpital Edouard Herriot, Pavillon H, F-69437 Lyon Cedex 3, France. Tel.: 33-4-72-11-75-50; Fax: 33-4-72-11-75-76; E-mail: abello@lyon151.inserm.fr.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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