Transcriptional activation and redox regulation of the tumor necrosis factor-alpha promoter in human T cells: role of the CRE/kappa3 promoter region.
Aberrancies in T cell expression of tumor necrosis factor-alpha (TNF-alpha) are frequently observed in inflammatory states characterized by oxidative stress due to excessive generation of hydrogen peroxide (H2O2) and other reactive oxygen species (ROS). In this study, we examined the possible effects of oxidative stress on the expression of TNF-alpha protein and transcriptional activation of the TNF-alpha promoter in human T cells. Results show that exposure of resting T cells to micromolar concentrations of H2O2 did not induce TNF-alpha protein production or transcriptional activation of the TNF-alpha promoter. However, oxidative signals resulted in a dose-dependent suppression of TNF-alpha protein production and transcriptional activation in T cells stimulated with the lectin phytohemagglutinin (PHA) and phorbol myristate acetate (PMA). Optimal suppression of TNF-alpha promoter activity was observed when cells were exposed to oxidative stress during early T cell activation, and other experiments demonstrated that the transactivation responses of the TNF-alpha promoter were quite susceptible to inhibition by both oxidative and reducing changes in cellular redox. Furthermore, reporter gene assays with 5' deletion mutants of the TNF-alpha promoter showed that the CRE/kappa3 composite site played a major role in activation of the TNF-alpha promoter by dual stimulatory signals and suppression of the TNF-alpha promoter by oxidative signals. Thus, T cell expression of TNF-alpha at the protein and transcriptional levels is highly regulated by changes in cellular redox, and the CRE/kappa3 composite site is important for both activation and redox regulation of the TNF-alpha promoter.
Liu
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Department of Medical Biochemistry, The William H. Davis Medical Research Center, The Ohio State University, Columbus 43210-1228, USA.
Whisler
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RL
D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D019169:Jurkat Cells; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010835:Phytohemagglutinins / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D013601:T-Lymphocytes / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
D010835:Phytohemagglutinins; D014409:Tumor Necrosis Factor-alpha; D006861:Hydrogen Peroxide; D013755:Tetradecanoylphorbol Acetate