Transcriptional regulation of glutamine synthetase gene expression by dexamethasone in L6 muscle cells.

Abstract Dexamethasone increases glutamine synthetase activity and mRNA abundance in L6 muscle cells in culture, apparently by a glucocorticoid receptor-mediated process. The data in this report reveal that increased glutamine synthetase mRNA abundance is attributable at least in part to an enhanced rate of transcription of the glutamine synthetase gene. "Nuclear runoff" assays of glutamine synthetase gene expression were performed with purified myonuclei from dexamethasone-treated or untreated L6 skeletal muscle cells. These assays showed glutamine synthetase transcription to be increased approximately 2-fold as early as 1 h after incubation of cells with dexamethasone (10(-7) M); there was no increase in the rate of transcription of the beta-tubulin gene, which served as a control. The increase in glutamine synthetase gene transcription correlates with increased glutamine synthetase enzymatic activity after dexamethasone treatment. Studies with actinomycin D indicated that the half-life of glutamine synthetase mRNA (7-8 h) is not altered by dexamethasone. Therefore, the degradation of glutamine synthetase mRNA is not affected by dexamethasone, and the increased glutamine synthetase mRNA level is attributable to increased transcription. The dexamethasone-mediated increase in glutamine synthetase mRNA abundance is glucocorticoid receptor-mediated; RU38486 (a glucocorticoid receptor blocker) completely blocked the effect of dexamethasone. The dexamethasone-mediated increase in glutamine synthetase gene transcription and steady-state mRNA level was not blocked by cycloheximide, indicating a direct effect.