Dexamethasone responsive element in the rat Na, K-ATPase beta1 gene coding region.
The Na, K-ATPase plays an essential role in active alveolar epithelial fluid resorption. In fetal and adult alveolar epithelial cells, glucocorticoids (GC) increase Na, K-ATPase activity, mRNA levels, and transcription rate of the beta(1) subunit. In this study, we describe a glucocorticoid responsive element (GRE) in the coding region of the rat Na, K-ATPase beta(1) gene in a rat lung epithelial cell line. Transient transfection experiments with the beta(1) subunit coding region with or without the 5' and 3' untranslated regions demonstrated responsiveness to dexamethasone induction and also identified a GRE at +434 in exon IV. The +434 GRE conferred dexamethasone responsiveness in a heterologous thymidine kinase promoter irrespective of its orientation to the beta(1) promoter. Transcriptional upregulation by dexamethasone was abolished in +434 mutants. Electrophoretic mobility shift assays (EMSA) demonstrated specific binding of nuclear proteins to the +434 GRE and the presence of the GC receptor. This specific binding was inhibited by a GRE previously described in the rat Na, K-ATPase beta(1) gene at -631. In conclusion, we identified a GRE at +434 in the exon IV of the rat Na, K-ATPase beta(1) gene.
Hao
Hong
H
Pulmonary and Critical Care Division, Department of Medicine, MMC 276, University of Minnesota, Minneapolis 55455, USA.
Rhodes
Richard
R
Ingbar
David H
DH
Wendt
Christine H
CH
D000818:Animals; D003907:Dexamethasone / Q000494:pharmacology*; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D005938:Glucocorticoids / Q000494:pharmacology*; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000187:drug effects* / Q000235:genetics
D005938:Glucocorticoids; D003907:Dexamethasone; D000254:Sodium-Potassium-Exchanging ATPase