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J. Biol. Chem., Vol. 275, Issue 31, 23627-23635, August 4, 2000
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From the Centro de Biología Molecular, Consejo Superior de
Investigaciones Científicas, Universidad
Autónoma de Madrid, Cantoblanco, E-28049 Madrid, Spain
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,
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 the
Ca2+/calcineurin pathway plays in COX-2
transcriptional regulation in T lymphocytes pointing to NFAT/activator
protein-1 transcription factors as essential for COX-2 promoter
regulation in these cells.
To whom correspondence should be addressed: Tel.: 34-91-3978413;
Fax: 34-91-3974799; E-mail: Mfresno@cbm.uam.es.
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