Roles of calcium and kinases in regulation of thrombin-stimulated preproendothelin-1 transcription.
The release of the vasoactive peptide endothelin-1 (ET-1) is Ca2+ dependent after thrombin stimulation; however, little is known about the pathways involved. We studied the importance of Ca(2+)-dependent signal transduction pathways on preproET-1 mRNA induction in human endothelial cells. Thrombin-mediated preproET-1 mRNA induction was inhibited after clamping of cytosolic free CA2+ concentration ([Ca2+]i) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. Chelation of extracellular Ca2+ with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid also had a significant inhibitory effect on the induction of preproET-1 mRNA. The Ca2+ ionophore A23187 induced constitutive as well as thrombin-stimulated preproET-1 mRNA expression. Mobilization of Ca2+ stores into the cytosol by inhibition of endoplasmic reticulum Ca(2+)-adenosinetriphosphatase with thapsigargin was effective also in inducing preproET-1 mRNA. Calmodulin antagonists W-7 and calmidazolium, as well as Ca2+/calmodulin-dependent kinase II inhibitor KN-62, significantly reduced thrombin-induced preproET-1 mRNA. Inhibition by cyclosporin A of the Ca(2+)-calmodulin-dependent phosphatase calcineurin potentiated constitutive preproET-1 mRNA. These data suggest that, in human endothelial cells, thrombin-mediated preproET-1 gene induction is regulated by a stimulatory Ca2+/calmodulin kinase II-dependent pathway.
Marsen
T A
TA
Department of Medicine, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA.
Simonson
M S
MS
Dunn
M J
MJ
D019703:Calcineurin; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D002147:Calmodulin / Q000502:physiology; D002148:Calmodulin-Binding Proteins / Q000037:antagonists & inhibitors; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D019332:Endothelin-1 / Q000235:genetics; D016232:Endothelins / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010770:Phosphotransferases / Q000502:physiology*; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000502:physiology*
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D002614:Chelating Agents; D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; D012333:RNA, Messenger; D010770:Phosphotransferases; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D013917:Thrombin; D002118:Calcium