title: Identification of a cis-element regulating transcriptional activity in response to fluid shear stress in bovine aortic endothelial cells.
abstract: Fluid shear stress exerts numerous effects on gene expression in endothelial cells. To investigate the regulatory mechanisms involved, we designed oligonucleotides composed of a 20-bp core containing the classical shear stress response element (SSRE+) or of a 20-bp core, in which base pairs flanking the SSRE were mutated (SSRE-). Hexamers of the oligonucleotides were cloned in front of reporter genes, transfected in bovine aortic endothelial cells (BAECs), and subjected to either a continuous low shear stress (3 dynes cm(-2)) or to a stepwise increase in shear stress from 3 dynes cm(-2) to 12 dynes cm(-2) (16/4 h). Shear stress increased reporter gene activity in cells transfected with pSSRE-, but not with pSSRE+. Cyclic strain (6%, 1 Hz, 4 h) did not significantly affect reporter gene activity. In gel retardation assays, more proteins bound to SSRE- than to SSRE+ in response to high shear stress. In competition experiments, a cAMP response element-binding (CREB) protein-specific oligonucleotide suppressed protein binding to the SSRE-; however, an antibody directed against CREB itself did not affect protein binding to the SSRE+. Our results indicate that the sequence ACC(G)/(T)AGACCAG represents a novel SSRE and that a protein that binds a CREB-specific oligonucleotide is part of the complex implicated in response to shear stress.
authors.affiliation: Institut für Kardiovaskuläre Physiologie, Klinikum der J. W. Goethe Universität, Frankfurt am Main, Germany. fisslthaler@em.uni-frankfurt.de
authors.forename: Beate
authors.initials: B
authors.lastname: Fisslthaler
authors.forename: Kerstin
authors.initials: K
authors.lastname: Boengler
authors.forename: Ingrid
authors.initials: I
authors.lastname: Fleming
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schaper
authors.forename: Rudi
authors.initials: R
authors.lastname: Busse
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Deindl
mesh_terms: D000818:Animals; D001011:Aorta; D001158:Arteries / Q000166:cytology; D001483:Base Sequence; D002417:Cattle; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D020574:Proto-Oncogene Proteins c-sis / Q000235:genetics / Q000378:metabolism; D020218:Response Elements; D013314:Stress, Mechanical; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: D004268:DNA-Binding Proteins; D020574:Proto-Oncogene Proteins c-sis; D014157:Transcription Factors