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Structure of the human 5-HT7 receptor gene and characterization of its promoter region.
Gene. 2007 Apr 15; 391(1-2):252-63.GENE

Abstract

The molecular mechanism regulating serotonin 5-HT(7) receptor expression is still unclear. In this study we provide evidence that transcription of the 5-HT(7) gene is at least partly regulated by Sp1 and Sp3. We isolated and sequenced >3000 bp of the upstream sequences and identified by RACE a number of transcriptional initiation sites over a region of 300 bp upstream of the coding region. This region has a high GC content, but contains no obvious TATA or a CAAT box. Besides a Sp1/Sp3 consensus motif, regulatory elements for AP2, Egr-1 and MAZ are also present. Transient transfection assays using deletion variants indicated that the GC-rich region is essential for full promoter activity. The role of Sp1 in this was confirmed by transient overexpression of both wild type Sp1 or dominant-negative forms. By gel shift and supershift analyses, targeting the Sp1 consensus sequence and the GC-rich region just upstream of the transcription initiation sites, binding of Sp1 and Sp3 was demonstrated. Both in vitro as well as in vivo experiments, using a cell line which endogenously expresses the 5-HT(7) receptor, indicated that mithramycin A, an inhibitor of Sp1/3 transcription factor binding, was able to inhibit 5-HT(7) promoter activity. Taken together these results support the essential role of Sp factors in regulating 5-HT(7) promoter activity.

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Authors+Show Affiliations

Laenen K
Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST), Department of Molecular Biology, Ghent University, UGent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium.
Haegeman G
No affiliation info available
Vanhoenacker P
No affiliation info available

MeSH

AnimalsBase SequenceBinding SitesBlotting, WesternCell LineCell Line, TumorElectrophoretic Mobility Shift AssayGC Rich SequenceGene ExpressionHumansLuciferasesMolecular Sequence DataPlicamycinPromoter Regions, GeneticProtein BindingReceptors, SerotoninRecombinant Fusion ProteinsReverse Transcriptase Polymerase Chain ReactionSequence AlignmentSequence Homology, Nucleic AcidSp1 Transcription FactorSp3 Transcription FactorTranscription Initiation SiteTransfection

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17321075

Citation

Laenen, Koen, et al. "Structure of the Human 5-HT7 Receptor Gene and Characterization of Its Promoter Region." Gene, vol. 391, no. 1-2, 2007, pp. 252-63.
Laenen K, Haegeman G, Vanhoenacker P. Structure of the human 5-HT7 receptor gene and characterization of its promoter region. Gene. 2007;391(1-2):252-63.
Laenen, K., Haegeman, G., & Vanhoenacker, P. (2007). Structure of the human 5-HT7 receptor gene and characterization of its promoter region. Gene, 391(1-2), 252-63.
Laenen K, Haegeman G, Vanhoenacker P. Structure of the Human 5-HT7 Receptor Gene and Characterization of Its Promoter Region. Gene. 2007 Apr 15;391(1-2):252-63. PubMed PMID: 17321075.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structure of the human 5-HT7 receptor gene and characterization of its promoter region. AU - Laenen,Koen, AU - Haegeman,Guy, AU - Vanhoenacker,Peter, Y1 - 2007/01/26/ PY - 2007/01/05/received PY - 2007/01/08/revised PY - 2007/01/08/accepted PY - 2007/2/27/pubmed PY - 2007/5/30/medline PY - 2007/2/27/entrez SP - 252 EP - 63 JF - Gene JO - Gene VL - 391 IS - 1-2 N2 - The molecular mechanism regulating serotonin 5-HT(7) receptor expression is still unclear. In this study we provide evidence that transcription of the 5-HT(7) gene is at least partly regulated by Sp1 and Sp3. We isolated and sequenced >3000 bp of the upstream sequences and identified by RACE a number of transcriptional initiation sites over a region of 300 bp upstream of the coding region. This region has a high GC content, but contains no obvious TATA or a CAAT box. Besides a Sp1/Sp3 consensus motif, regulatory elements for AP2, Egr-1 and MAZ are also present. Transient transfection assays using deletion variants indicated that the GC-rich region is essential for full promoter activity. The role of Sp1 in this was confirmed by transient overexpression of both wild type Sp1 or dominant-negative forms. By gel shift and supershift analyses, targeting the Sp1 consensus sequence and the GC-rich region just upstream of the transcription initiation sites, binding of Sp1 and Sp3 was demonstrated. Both in vitro as well as in vivo experiments, using a cell line which endogenously expresses the 5-HT(7) receptor, indicated that mithramycin A, an inhibitor of Sp1/3 transcription factor binding, was able to inhibit 5-HT(7) promoter activity. Taken together these results support the essential role of Sp factors in regulating 5-HT(7) promoter activity. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/17321075/Structure_of_the_human_5_HT7_receptor_gene_and_characterization_of_its_promoter_region_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-1119(07)00040-6 DB - PRIME DP - Unbound Medicine ER -