Thrombin regulation of endothelin-1 gene in isolated human pulmonary endothelial cells.
Endothelin (ET)-1 is a potent vasoconstrictor elicited from endothelial cells in response to a variety of stimuli and an important mediator for a variety of vascular diseases including pulmonary hypertension. In this paper, we describe the molecular regulation of the ET-1 gene in response to a vasoactive mediator, thrombin, in human pulmonary endothelial cells. Thrombin induces preproET-1 mRNA through a transcriptionally dependent mechanism, with a peak induction after 1 h of exposure. Analysis of chromatin structure identified several DNase I-hypersensitive regions under both basal and thrombin-stimulated conditions that reside in the 5'-promoter region, indicating that the ET-1 promoter is a constitutive promoter. Deletion analysis was employed as a functional assay to identify regions of the ET-1 promoter that are important in transcriptional regulation. We found that sites between -141 and -378 bp are essential for basal activity and that those between -378 and -484 bp are essential for thrombin-stimulated activity. However, full expression under both conditions required an element(s) within -952 bp.
Golden
C L
CL
Department of Pediatrics, University of Florida, Gainesville 32610, USA.
Nick
H S
HS
Visner
G A
GA
D002469:Cell Separation; D002478:Cells, Cultured; D002843:Chromatin / Q000502:physiology; D019332:Endothelin-1 / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D017353:Gene Deletion; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D011401:Promoter Regions, Genetic / Q000502:physiology; D011651:Pulmonary Artery / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D013917:Thrombin / Q000502:physiology*
D002843:Chromatin; D019332:Endothelin-1; D012333:RNA, Messenger; D013917:Thrombin