The serotonin 5-HT4 receptor (where 5-HT stands for 5-hydroxy-tryptamine) is a member of the seven transmembrane-spanning G-protein-coupled family of receptors and mediates many cellular functions both in the central nervous system and at the periphery. In the present study, we isolated and characterized the 5′-flanking region of the h5-HT4 (human 5-HT4) receptor. We demonstrate the existence of a novel exon that corresponds to the 5′-untranslated region of the h5-HT4 receptor gene. RNase protection analysis and reverse transcriptase–PCR experiments performed on human atrial RNA demonstrated that the major transcription start site of the h5-HT4 receptor gene is located at −3185 bp relative to the first ATG codon. In addition, a 1.2 kb promoter fragment which drives the transcription of the 5-HT4 receptor was characterized. The promoter region lacks TATA and CAAT canonical motifs in the appropriate location, but contains putative binding sites for several transcription factors. Transient transfection assays revealed that the (−3299/−3050) gene fragment possesses the ability to promote the expression of the luciferase reporter gene in human cell lines. In contrast, the promoter was silent in monkey COS-7 cells, indicating the requirement of specific factors to initiate transcription in human cells. In addition to the promoter element, enhancer activity was found in a region (−220/−61) located in the long 5′-untranslated region. Mutational analysis, gel shift and transfection assays identified an Nkx2.5 (NK2-transcription-factor-related 5)-like binding site as a regulatory sequence of this enhancer. Our results suggest a complex regulation of the h5-HT4 receptor gene expression involving distinct promoters and non-coding exons.
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Research Article|
April 05 2005
Functional studies of the 5′-untranslated region of human 5-HT4 receptor mRNA
Marjorie MAILLET;
Marjorie MAILLET
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Monique GASTINEAU;
Monique GASTINEAU
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Pascal BOCHET;
Pascal BOCHET
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Marie-Liesse ASSELIN-LABAT;
Marie-Liesse ASSELIN-LABAT
†INSERM U-461, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Eric MOREL;
Eric MOREL
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Jean-Noël LAVERRIÈRE;
Jean-Noël LAVERRIÈRE
‡CNRS UMR 7079, Université Pierre et Marie Curie, F-75270, Paris, France
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Anne-Marie LOMPRÉ;
Anne-Marie LOMPRÉ
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Rodolphe FISCHMEISTER;
Rodolphe FISCHMEISTER
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
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Frank LEZOUALC'H
Frank LEZOUALC'H
1
*Cardiologie Cellulaire et Moléculaire, INSERM U-446, IRF-75, Université Paris-Sud, Faculté de Pharmacie, F-92296 Châtenay-Malabry, France
1To whom correspondence should be addressed (email Frank.Lezoualch@cep.u-psud.fr).
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Publisher: Portland Press Ltd
Received:
May 06 2004
Revision Received:
November 23 2004
Accepted:
December 02 2004
Accepted Manuscript online:
December 02 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 387 (2): 463–471.
Article history
Received:
May 06 2004
Revision Received:
November 23 2004
Accepted:
December 02 2004
Accepted Manuscript online:
December 02 2004
Citation
Marjorie MAILLET, Monique GASTINEAU, Pascal BOCHET, Marie-Liesse ASSELIN-LABAT, Eric MOREL, Jean-Noël LAVERRIÈRE, Anne-Marie LOMPRÉ, Rodolphe FISCHMEISTER, Frank LEZOUALC'H; Functional studies of the 5′-untranslated region of human 5-HT4 receptor mRNA. Biochem J 15 April 2005; 387 (2): 463–471. doi: https://doi.org/10.1042/BJ20040744
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