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          Institute: MPI für Herz- und Lungenforschung (W. G. Kerckhoff Institut)     Collection: Publikationen des W. G. Kerckhoff-Instituts     Display Documents



  history
ID: 199054.0, MPI für Herz- und Lungenforschung (W. G. Kerckhoff Institut) / Publikationen des W. G. Kerckhoff-Instituts
Identification of a cis-element regulating transcriptional activity in response to fluid shear stress in bovine aortic endothelial cells
Authors:Fisslthaler, B.; Boengler, K.; Fleming, I.; Schaper, W.; Busse, R.; Deindl, E.
Date of Publication (YYYY-MM-DD):2003
Title of Journal:Endothelium
Volume:10
Issue / Number:4-5
Start Page:267
End Page:275
Review Status:not specified
Audience:Not Specified
Abstract / Description: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.
Free Keywords:Animals
; Aorta
; Arteries/cytology
; Base Sequence
; Cattle
; DNA-Binding Proteins/genetics/metabolism
; Endothelial Cells/metabolism
; Endothelium, Vascular/cytology/*metabolism
; Enhancer Elements (Genetics)
; Genes, Reporter
; Genetic Vectors
; Molecular Sequence Data
; Plasmids
; Promoter Regions (Genetics)
; Proto-Oncogene Proteins c-sis/genetics/metabolism
; *Response Elements
; Stress, Mechanical
; Support, Non-U.S. Gov't
; *Trans-Activation (Genetics)
; Transcription Factors/genetics/metabolism
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für physiologische und klinische Forschung
Identifiers:URL:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=... [ID No:1]
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