title: Post-transcriptional divergence in the regulation of CAT-2A, CAT-2B and iNOS expression by dexamethasone in vascular smooth muscle cells.
abstract: Upregulation of L-arginine transport by pro-inflammatory mediators is a widely reported phenomenon which accompanies the expression of the inducible nitric oxide synthase (iNOS) enzyme in various cells. Both processes require de novo protein synthesis which may be regulated differentially through diverging signalling pathways. This is particularly defined by observations that the glucocorticoid dexamethasone, acting potentially through NF-κB, selectively blocks the expression of iNOS whilst having little or no effect on transport; suggesting that this ubiquitous transcription factor may not be required for induced transporter activity. This notion is however controversial as is the suggestion that dexamethasone may regulate iNOS expression exclusively through NF-κB. Thus, to further understand the mechanisms that control these processes, we have examined the level at which dexamethasone acts, investigating whether this involves NF-κB and whether the latter selectively regulates iNOS induction. Our current data directly demonstrate that induced L: -arginine transport is critically dependent on the activation of NF-κB, and further confirmed its role in the induction of iNOS in rat aortic smooth muscle cells. More importantly, dexamethasone enhanced both iNOS and CAT gene expression but repressed iNOS protein with no noticeable effects on transporter function or indeed NF-κB activation. These novel and unexpected findings reflect the complex nature of the regulation of iNOS by glucocorticoids and prove, contrary to previous assumptions, that dexamethasone can regulate CAT gene expression despite failing to alter transporter function. Moreover, the effects of dexamethasone occur through a non-NF-κB-mediated action even though NF-κB is required for both processes.
authors: School of Life Sciences, University of Hertfordshire, College Lane, Hatfield AL10 9AB, UK. s.n.1.thakur@herts.ac.uk
authors: Shori
authors: S
authors: Thakur
authors: Anwar R
authors: AR
authors: Baydoun
mesh_terms: D000818:Animals; D001120:Arginine / Q000378:metabolism; D027201:Cationic Amino Acid Transporter 1 / Q000235:genetics / Q000378:metabolism; D027202:Cationic Amino Acid Transporter 2 / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D005786:Gene Expression Regulation; D005938:Glucocorticoids / Q000494:pharmacology*; D020672:I-kappa B Proteins / Q000378:metabolism; D018836:Inflammation Mediators / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D061988:Proteasome Inhibitors; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D027201:Cationic Amino Acid Transporter 1; D027202:Cationic Amino Acid Transporter 2; D005938:Glucocorticoids; D020672:I-kappa B Proteins; D018836:Inflammation Mediators; D007976:Leupeptins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606206:Nfkbia protein, rat; D061988:Proteasome Inhibitors; D000072000:NF-KappaB Inhibitor alpha; D009569:Nitric Oxide; D003907:Dexamethasone; D007371:Interferon-gamma; D001120:Arginine; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde