UTP Induces Osteopontin Expression through a Coordinate Action of NFκB, Activator Protein-1, and Upstream Stimulatory Factor in Arterial Smooth Muscle Cells
| Autor: | Isabelle Belloc, Mylène Potier, Lodewijk V. Dekker, Elisabeth Génot, Marie-Ange Renault, Alain-Pierre Gadeau, Sandra Jalvy, Claude Desgranges |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
MAPK/ERK pathway
Transcription Genetic Uridine Triphosphate IκB kinase Polymerase Chain Reaction Biochemistry Upstream Stimulatory Factor chemistry.chemical_compound Cell Movement Luciferases Promoter Regions Genetic Cells Cultured Genes Dominant Uridine triphosphate Mitogen-Activated Protein Kinase 1 Mitogen-Activated Protein Kinase 3 NF-kappa B Arteries Up-Regulation DNA-Binding Proteins Signal transduction Plasmids Protein Binding Signal Transduction Sialoglycoproteins Blotting Western Biology Models Biological DNA-binding protein stomatognathic system Animals Electrophoretic mobility shift assay Rats Wistar Molecular Biology Binding Sites NADPH Oxidases Muscle Smooth DNA Cell Biology Molecular biology Rats Transcription Factor AP-1 Gene Expression Regulation chemistry Mutation Mutagenesis Site-Directed Upstream Stimulatory Factors Osteopontin Chromatin immunoprecipitation Gene Deletion Transcription Factors |
| Zdroj: | Journal of Biological Chemistry. 280:2708-2713 |
| ISSN: | 0021-9258 |
| Popis: | Osteopontin (OPN) is an important chemokinetic agent for several cell types. Our earlier studies have shown that its expression is essential for uridine triphosphate (UTP)-mediated migration of vascular smooth muscle cells. We demonstrated previously that the activation of an AP-1 binding site located 76 bp upstream of the transcription start in the rat OPN promoter is involved in the induction of OPN expression. In this work, using a luciferase promoter deletion assay, we identified a new region of the rat OPN promoter (-1837 to -1757) that is responsive to UTP. This region contains an NFkappaB site located at -1800 and an Ebox located at -1768. Supershift electrophoretic mobility shift assay and chromatin immunoprecipitation assays identified NFkappaB and USF-1/USF-2 as the DNA binding proteins induced by UTP, respectively, for these two sites. Using dominant negative mutants of IkappaB kinase and USF transcription factors, we confirmed that NFkappaB and USF-1/USF-2 are involved in the UTP-mediated expression of OPN. Using a pharmacological approach, we demonstrated that USF proteins are regulated by the extracellular signal-regulated kinase (ERK)1/2 pathway, just as the earlier discovered AP-1 complex, whereas NFkappaB is up-regulated through PKCdelta signals. Finally, our work suggests that the UTP-stimulated OPN expression involves a coordinate regulation of PKCdelta-NFkappaB, ERK1/2-USF, and ERK1/2/NAD(P)H oxidase AP-1 signaling pathways. |
| Databáze: | OpenAIRE |
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