title: Antagonizing effect of AP-1 on glucocorticoid induction of urea cycle enzymes: a study of hyperammonemia in carnitine-deficient, juvenile visceral steatosis mice.
abstract: Hyperammonemia is one of the major symptoms of primary carnitine deficiency. Carnitine-deficient juvenile visceral steatosis (JVS) mice show hyperammonemia during the weaning period. We have found that all of the urea cycle enzyme genes are suppressed and that N-acetylglutamate, an allosteric activator of the first step enzyme of the urea cycle, carbamoyl phosphate synthetase I (CPS), is not deficient in the liver of JVS mice. Induction of the urea cycle enzymes by glucocorticoid in rat primary cultured hepatocytes was suppressed by the addition of long-chain fatty acids. The suppression of the urea cycle enzyme genes in vivo and in vitro is accompanied by stimulated AP-1 DNA-binding activity. However, mRNA of phosphoenolpyruvate carboxykinase, one of the gluconeogenic enzymes which responds to glucocorticoid, is further stimulated by the addition of fatty acid. From these results, we postulate that protein-protein interaction between glucocorticoid receptors and AP-1 is not the major mechanism of suppression, but that AP-1 causes the suppression through a cis-element on the gene. After cloning promoter and enhancer regions of the mouse CPS gene and comparing rat and mouse, we found that an AP-1 site was present just 3'-downstream of the minimal essential enhancer fragment previously described. We also found that the presence of an AP-1 site in reporter gene constructs resulted in suppression of the reporter genes in the liver of carnitine-deficient JVS mice and suppression of glucocorticoid induction by long-chain fatty acid in cultured hepatocytes.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, 890-8520, Japan. takesah@med2.kufm.kagoshima-u.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Saheki
authors.forename: M X
authors.initials: MX
authors.lastname: Li
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000235:genetics; D002331:Carnitine / Q000172:deficiency*; D005227:Fatty Acids / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005938:Glucocorticoids / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology; D022124:Hyperammonemia / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D020218:Response Elements / Q000235:genetics; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism*; D014508:Urea / Q000378:metabolism*
chemical_list: D005227:Fatty Acids; D005938:Glucocorticoids; D018808:Transcription Factor AP-1; D014508:Urea; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D002331:Carnitine