title: UTP induces osteopontin expression through a coordinate action of NFkappaB, activator protein-1, and upstream stimulatory factor in arterial smooth muscle cells.
abstract: 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.
authors.affiliation: INSERM, U441 Athérosclérose, Bordeaux 33600, France.
authors.forename: Marie-Ange
authors.initials: MA
authors.lastname: Renault
authors.forename: Sandra
authors.initials: S
authors.lastname: Jalvy
authors.forename: Mylène
authors.initials: M
authors.lastname: Potier
authors.forename: Isabelle
authors.initials: I
authors.lastname: Belloc
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Genot
authors.forename: Lodewijk V
authors.initials: LV
authors.lastname: Dekker
authors.forename: Claude
authors.initials: C
authors.lastname: Desgranges
authors.forename: Alain-Pierre
authors.initials: AP
authors.lastname: Gadeau
mesh_terms: D000818:Animals; D001158:Arteries / Q000473:pathology*; D001665:Binding Sites; D015153:Blotting, Western; D002465:Cell Movement; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D017353:Gene Deletion; D005786:Gene Expression Regulation; D005799:Genes, Dominant; D008156:Luciferases / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D008954:Models, Biological; D009130:Muscle, Smooth / Q000166:cytology*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D019255:NADPH Oxidases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D053495:Osteopontin; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics; D015398:Signal Transduction; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D015854:Up-Regulation; D051720:Upstream Stimulatory Factors; D014544:Uridine Triphosphate / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D016328:NF-kappa B; D012795:Sialoglycoproteins; C506897:Spp1 protein, rat; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D051720:Upstream Stimulatory Factors; C497182:Usf1 protein, rat; D053495:Osteopontin; D004247:DNA; D008156:Luciferases; D019255:NADPH Oxidases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D014544:Uridine Triphosphate