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01.05.2004 | Article

Protein hydrolysates stimulate proglucagon gene transcription in intestinal endocrine cells via two elements related to cyclic AMP response element

verfasst von: J.-C. Gevrey, M. Malapel, J. Philippe, G. Mithieux, J.-A. Chayvialle, J. Abello, M. Cordier-Bussat

Erschienen in: Diabetologia | Ausgabe 5/2004

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Abstract

Aims/hypothesis

Protein hydrolysates (peptones) increase not only glucagon-like peptide-1 (GLP-1) secretion but also transcription of the proglucagon (PG) gene in the intestine. The critical physiological roles of gut-derived GLPs raised hope for their therapeutic use in several disorders, especially GLP-1 in diabetes. We aimed to investigate the molecular mechanisms involved in this nutrient–PG gene interaction.

Methods

Wild-type and mutated PG promoter fragments fused to the luciferase reporter gene were transfected into enteroendocrine STC-1 cells, which were then either treated or not with peptones. Co-transfection with expression vectors of dominant-negative forms of cAMP response element binding protein (CREB) and protein kinase A (PKA) proteins were performed, as well as electrophoresis mobility shift assays.

Results

Deletion analysis showed that the promoter region spanning between −350 and −292 bp was crucial for the transcriptional stimulation induced by peptones. Site-directed mutagenesis of the canonical cAMP response element (CRE_PG) and of the adjacent putative CRE site (CRE-like1) led to a dramatic inhibition of the promoter responsiveness to peptones. Over expression of a dominant-negative mutant of CREB or of PKA produced a comparable and selective inhibitory effect on the activity of transfected promoter fragment containing the −350/−292 sequence. EMSA showed that CREB and fra2 transcription factors bound to CRE_PG and CRE-like1 elements respectively, independently of peptone treatment.

Conclusions/interpretation

Our report identified cis- and trans-regulatory elements implicated in the transcriptional control of PG gene by nutrients in enteroendocrine cells. It highlights the role of a previously unsuspected CRE-like1 element, and emphasises the importance of CRE-related sequences in the regulation of PG gene transcription in the intestine.

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Titel: Protein hydrolysates stimulate proglucagon gene transcription in intestinal endocrine cells via two elements related to cyclic AMP response element
verfasst von: J.-C. Gevrey
M. Malapel
J. Philippe
G. Mithieux
J.-A. Chayvialle
J. Abello
M. Cordier-Bussat
Publikationsdatum: 01.05.2004
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 5/2004
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-004-1380-0

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