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A more recent version of this article appeared on May 17, 2002
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M110013200v1
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Papers In Press, published online ahead of print March 13, 2002
J. Biol. Chem, 10.1074/jbc.M110013200
Submitted on October 17, 2001
Revised on February 20, 2002
Accepted on March 13, 2002

IL-13 gene expression is regulated by GATA-3 in T cells: Role of a critical association of a GATA and two GATG motifs

Cécile Lavenu-Bombled, Cecelia D. Trainor, Iman Makeh, Paul-Henri Roméo, and Isabelle Max-Audit

Unité 474, INSERM, Paris 75014

Corresponding Author: maxaudit{at}cochin.inserm.fr

Using a transgenic approach, we studied the role of GATA-3 in T cells. As previously shown, enforced GATA-3 expression in transgenic mice inhibits Th1 differentiation of CD4 T cells but unexpectedly, both type 1 (IFNgamma) and type 2 (IL-4 and IL-13) cytokine genes were activated in the transgenic CD8 T cells. As IL-13 gene expression was highly enhanced in vivo by GATA-3 expression, we studied the human and the mouse IL-13 gene promoters and found an evolutionary conserved association of a consensus GATA binding site and two GATG motifs. We showed that efficient GATA-3 binding to this regulatory sequence required these three motifs and that the affinity of the GATA zinc fingers for this association was five times higher than for the consensus GATA binding site alone. Transfections in a T cell line or transactivation by GATA-3 showed that the combination of the three sites was required for full transcriptional activity of the IL-13 gene promoter. Finally we showed that this association of binding sites causes a very high sensitivity of the IL-13 gene promoter to small variations in the level of GATA-3 protein. Altogether, these results indicate an important role of GATA-3 in CD8 cytokine gene expression and demonstrate that a critical network of GATA binding sites highly modulates GATA-3 activity.


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