Transcriptional regulation of the tissue-type plasminogen-activator gene in human endothelial cells: identification of nuclear factors that recognise functional elements in the tissue-type plasminogen-activator gene promoter

Détails

ID Serval
serval:BIB_344FBCBCEC63
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Transcriptional regulation of the tissue-type plasminogen-activator gene in human endothelial cells: identification of nuclear factors that recognise functional elements in the tissue-type plasminogen-activator gene promoter
Périodique
European Journal of Biochemistry
Auteur(s)
Costa M., Shen Y., Maurer F., Medcalf R. L.
ISSN
0014-2956 (Print)
Statut éditorial
Publié
Date de publication
11/1998
Volume
258
Numéro
1
Pages
123-31
Notes
Journal Article Research Support, Non-U.S. Gov't --- Old month value: Nov 15
Résumé
The gene encoding human tissue-type plasminogen activator (t-PA) is regulated in a cell-type-specific manner. Previous studies in non-endothelial cells have indicated that basal and phorbol ester mediated induction is controlled by a cAMP response element (CRE) referred to as the tPACRE, and an activating protein 2 (AP-2)-like site. The classification of the AP-2-like site was assigned on the basis of its sequence homology, but has been shown in some cell systems to be recognised by promoter-specific transcription factor-1 (Sp-1). Here, we have investigated the transcriptional regulation of the t-PA gene in endothelial cells and addressed the functional roles of the tPACRE and the Sp-1/AP-2-like sites. 5'-RACE experiments indicate that the t-PA gene uses two transcription initiation sites in these cells with the downstream site being preferred. Functional analyses of the t-PA promoter using reporter-gene constructs transfected into C11STH endothelial cells demonstrate that the first 410 bp of the t-PA promoter confers an increase in reporter-gene activity on treatment with 4beta-phorbol 12-myristate 13-acetate (PMA). Mutagenesis of either the tPACRE or the Sp-1/AP-2 site weakens both basal and inducible expression, while disruption of both sites renders the promoter completely unresponsive. Using supershift assays, we identify the predominant tPACRE-binding proteins in nuclear extracts prepared from both C11STH cells and primary umbilical vein endothelial cells (HUVECs) as activating transcription factor 2, CREB (cAMP-responsive-element-binding protein), CREM (cAMP response element modulator) and c-jun. Treatment of cells with PMA results in a selective recruitment of jun-D to the tPACRE, while Sp-1 was identified as the major transcription factor that recognises the AP-2-like site. Based on this data and previous reports, we have reassigned this as a Sp-1-binding site. Finally, the identification of specific endothelial-derived t-PACRE-binding proteins suggests an integral role for these factors in the regulation of t-PA gene expression in human endothelial cells.
Mots-clé
Base Sequence Cell Line, Transformed Cell Nucleus/metabolism DNA Primers *Gene Expression Regulation/drug effects Humans *Promoter Regions (Genetics) RNA, Messenger/genetics Tetradecanoylphorbol Acetate/pharmacology Tissue Plasminogen Activator/*genetics *Transcription, Genetic/drug effects Tumor Necrosis Factor-alpha/pharmacology
Pubmed
Web of science
Open Access
Oui
Création de la notice
25/01/2008 15:19
Dernière modification de la notice
08/05/2019 16:52
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