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Papers In Press, published online ahead of print May 17, 2000
Centro de Biologia Molecular CSIC-UAM, Cantoblanco, Madrid 28049
Corresponding Author: mainiguez{at}cbm.uam.es
We have previously reported that transcriptional induction of Cyclooxygenase-2 (Cox-2) isoenzyme occurs early after T cell receptor triggering, suggesting functional implications of cyclooxygenase activity in this process. Here, we identify the cis-acting elements responsible for the transcriptional activation of this gene in human T lymphocytes. Cox-2 promoter activity was induced upon T cell activation both in primary resting T lymphocytes and in Jurkat cells. This induction was abrogated by inhibition of calcineurin phosphatase with the immunosuppressive drug Cyclosporin A (CsA), whereas expression of an active calcineurin catalytic subunit enhanced Cox-2 transcriptional activation. Moreover, cotransfection of nuclear factor of activated T cells (NFAT) wild type protein transactivated Cox-2 promoter activity. Conversely, dominant negative mutants of NFATc or c-Jun proteins inhibited Cox-2 induction. Electrophoretic mobility shift assays and site-directed mutagenesis allowed the identification of two regions of DNA located in the positions -117 and -58 relative to the transcriptional start site that serves as NFAT recognition sequences. These results emphasize the central role that Ca++/calcineurin pathway plays in Cox-2 transcriptional regulation in T lymphocytes pointing to NFAT/AP-1 transcription factors as essential for Cox-2 promoter regulation in these cells.
J. Biol. Chem, 10.1074/jbc.M001381200
Submitted on February 18, 2000
Revised on May 4, 2000
Accepted on May 17, 2000
An essential role of NFAT in the regulation of the expression of cyclooxygenase-2 gene in human T lymphocytes
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