Transcriptional regulation of gamma-glutamylcysteine synthetase-heavy subunit by oxidants in human alveolar epithelial cells

Biochemical and Biophysical Research Communications
I RahmanC A Smith

Abstract

We studied the regulation of glutathione (GSH) synthesis and characterised the 5'-promoter region of the gamma-glutamylcysteine synthetase-heavy subunit (gammaGCS-HS) gene in human alveolar type II cells (A549) following exposure to menadione (MQ) and hydrogen peroxide (H2O2). Both MQ (100 microM) and H2O2 (100 microM) exposure increased intracellular GSH levels associated with increased gammaGCS activity. This was concomitant with enhanced expression of gammaGCS-HS mRNA. Transfection of deletion constructs of the gammaGCS-HS promoter (-1050 to +82 bp) in a chloramphenicol acetyl transferase (CAT) reporter system revealed that an human antioxidant response element (hARE), present within the proximal region of the promoter (-1050 to -818 bp), is not required for oxidant-mediated gene induction. We conclude that oxidant stress-induced gammaGCS-HS mRNA expression is associated with AP-1 or AP-1 like responsive elements (-817 to +45 bp).

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