Functional studies of the 5′-untranslated region of human 5-HT 4 receptor mRNA

Marjorie Maillet, Monique Gastineau, Pascal Buchet, Marie-Liesse Labat, Eric Morel, Jean Noël Laverrière, Anne Marie Lompré, Rodolphe Fischmeister, Frank Lezoualc'h

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8 Scopus citations

Abstract

The serotonin 5-HT4 receptor (where 5-HT stands for 5-hydroxytryptamine) 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.

Original languageEnglish (US)
Pages (from-to)463-471
Number of pages9
JournalBiochemical Journal
Volume387
Issue number2
DOIs
StatePublished - Apr 15 2005
Externally publishedYes

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Keywords

  • 5′-Untranslated region
  • 5-Hydroxytryptamine (serotonin)
  • CAMP
  • MRNA
  • Nkx2.5
  • Transcriptional regulation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Maillet, M., Gastineau, M., Buchet, P., Labat, M-L., Morel, E., Laverrière, J. N., Lompré, A. M., Fischmeister, R., & Lezoualc'h, F. (2005). Functional studies of the 5′-untranslated region of human 5-HT 4 receptor mRNA. Biochemical Journal, 387(2), 463-471. https://doi.org/10.1042/BJ20040744