TY - JOUR
T1 - Divergent effects of dithiocarbamates on AP-1-containing and AP-1-less NFAT sites
AU - Saccani, Simona
AU - Saccani, Alessandra
AU - Varesio, Luigi
AU - Ghosh, Paritosh
AU - Young, Howard A.
AU - Sica, Antonio
PY - 1999
Y1 - 1999
N2 - Changes in the redox status of cells affect NF-κB and activator protein (AP)-1 nuclear expression and activity. In particular, antioxidants decrease NF-κB and increase AP-1 transcriptional activity, thereby regulating gene expression. In T cells, low concentrations of antioxidants enhance IL-2 and inhibit IL-4 expression. Since NFAT binding sites play an essential role in regulating IL-2 and IL-4 gene transcription, we studied the effects of dithiocarbamates, using the pyrrolidine derivative of dithiocarbamate (PDTC), on the activity of the distinct AP-1-containing IL-2 NFAT and AP-1-less IL-4 NFAT enhancers elements. Consistent with the presence of AP-1 proteins within the IL-2 NFAT complex, PDTC strongly enhanced phorbol 12-myristate 13-acetate/phytohemagglutinin-induced NFAT binding to the IL-2 NFAT enhancer and transcriptional activity of a reporter plasmid driven by this NFAT enhancer. In contrast, the activity of the IL-4 NFp enhancer, which does not bind AP-1, was abolished by PDTC treatment. In the Jurkat T cell line treated with PDTC, co-expression of the Ca2+/calmodulin-dependent phosphatase, calcineurin, completely restored the IL-4 NFp enhancer activity. Our data indicate that calcineurin-mediated NFAT activity is a target for antioxidants and provides new insights into the molecular mechanisms controlling differential cytokine gene expression.
AB - Changes in the redox status of cells affect NF-κB and activator protein (AP)-1 nuclear expression and activity. In particular, antioxidants decrease NF-κB and increase AP-1 transcriptional activity, thereby regulating gene expression. In T cells, low concentrations of antioxidants enhance IL-2 and inhibit IL-4 expression. Since NFAT binding sites play an essential role in regulating IL-2 and IL-4 gene transcription, we studied the effects of dithiocarbamates, using the pyrrolidine derivative of dithiocarbamate (PDTC), on the activity of the distinct AP-1-containing IL-2 NFAT and AP-1-less IL-4 NFAT enhancers elements. Consistent with the presence of AP-1 proteins within the IL-2 NFAT complex, PDTC strongly enhanced phorbol 12-myristate 13-acetate/phytohemagglutinin-induced NFAT binding to the IL-2 NFAT enhancer and transcriptional activity of a reporter plasmid driven by this NFAT enhancer. In contrast, the activity of the IL-4 NFp enhancer, which does not bind AP-1, was abolished by PDTC treatment. In the Jurkat T cell line treated with PDTC, co-expression of the Ca2+/calmodulin-dependent phosphatase, calcineurin, completely restored the IL-4 NFp enhancer activity. Our data indicate that calcineurin-mediated NFAT activity is a target for antioxidants and provides new insights into the molecular mechanisms controlling differential cytokine gene expression.
KW - AP-1
KW - NF-κB
KW - NFAT
KW - Pyrrolidine derivative of dithiocarbamate
UR - http://www.scopus.com/inward/record.url?scp=0032964813&partnerID=8YFLogxK
U2 - 10.1002/(SICI)1521-4141(199904)29:04<1194::AID-IMMU1194>3.0.CO;2-0
DO - 10.1002/(SICI)1521-4141(199904)29:04<1194::AID-IMMU1194>3.0.CO;2-0
M3 - Article
SN - 0014-2980
VL - 29
SP - 1194
EP - 1201
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 4
ER -