Tyrosine-kinase-dependent regulation of the nitric oxide synthase gene by endothelin-1 in human endothelial cells

Abstract

In vascular endothelium, endothelium-derived relaxing factor, predominantly nitric oxide (NO), is synthesized by endothelial NO synthase (eNOS). While regulatory influences on eNOS enzyme activity are widely clarified, little is known about the regulation of the eNOS gene. We investigated the regulatory signaling mechanisms of eNOS mRNA expression and accumulated NO production in human endothelial cells. Northern blot analysis and NO assays demonstrate that the vasoconstrictor peptide endothelin-1 (ET-1) induces the eNOS gene and leads to accumulated NO production. Induction occurs via ETA receptor activation and depends on improved transcript stability. It is maintained for incubation periods of 30-90 min and tapers thereafter. Regulatory signaling mechanisms depend on de novo protein synthesis to control eNOS mRNA fate. Selectively blocking protein tyrosine kinases (PTK) and inhibiting protein kinase C (PKC) inhibit eNOS mRNA expression and accumulated NO secretion. These observations indicate that regulation of eNOS at the genomic level occurs via post-transcriptional mechanisms. Two protein-bound intracellular kinase pathways, PTK and PKC, regulate eNOS mRNA expression and accumulated NO production.

Contribution of the endothelin and renin–angiotensin systems to the vascular changes in rats chronically treated with ouabain

“…However, there is evidence suggesting that activation of ET A receptors also stimulates NO release. Marsen et al . (1999) showed that ET‐1 acting on ET A receptors induces eNOS gene expression via tyrosine kinase‐ and protein kinase‐dependent pathways.…”

β-Adrenergic receptors and nitric oxide generation in the cardiovascular system

“…More recently, it has been shown that adenosine, via the A 2a receptor, induces NO production in HUVECs and that this is mediated via the MEK1/2/ERK1/2 pathway [48]. There is also evidence that, in HUVECs, endothelin-induced NOS 3 regulation is tyrosine kinase-dependent [49], which is of relevance since ERK1/2 is a tyrosine kinase; furthermore, bAR-mediated PKA signalling can activate MAPK through switching of the coupling of bARs from a G s protein to a G i protein as a consequence of PKA-mediated b 2 AR phosphorylation [50]. PKA itself has also been shown to activate the MAPK cascade through direct stimulation of Rap 1 [51].…”

Possible interactions of the endothelial constitutive nitric oxide synthase genotype with alcohol drinking and walking time for high serum uric acid levels among Japanese

“…However, it was reported that ethanol induced a rapid increase of ecNOS protein and mRNA expression levels and activity [44][45][46][47][48]. Ethanol modulates tyrosine kinase activity [49], and tyrosine kinase regulates expression of ecNOS via posttranscriptional mechanisms [50,51]. In addition, ethanol dose dependently increased basal ecNOS activity via a mechanism involving a pertussis toxin-sensitive G protein in the absence of any effect on cell viability or NOS protein expression [46].…”