Interaction of NF-κB and NFAT with the interferon-γ promoter

Antonio Sica, Linda Dorman, Vincenzo Viggiano, Marco Cippitelli, Paritosh Ghosh, Nancy Rice, Howard A. Young

Research output: Contribution to journalArticle

Abstract

Interferon-γ (IFN-γ) is a pleiotropic lymphokine whose production is restricted to activated T cells and NK cells. Along with other cytokines, IFN-γ gene expression is inhibited by the immunosuppressant cyclosporin A. We have previously identified an intronic enhancer region (C3) of the IFN-γ gene that binds the NF-≃B protein c-Rel and that shows partial DNA sequence homology with the cyclosporin A-sensitive NFAT binding site and the 3'-half of the NF-≃B consensus site. Sequence analysis of the IFN-γ promoter revealed the presence of two additional C3-related elements (C3-1P and C3- 3P). In addition, an NF-≃B site (IFN-γ ≃B) was identified within the promoter region. Based on this observation, we have analyzed the potential role of NF-≃B and NFAT family members in regulating IFN-γ transcription. Electrophoretic mobility shift assay analysis demonstrated that after T cell activation, the p50 and p65 NF-≃B sub-units bind specifically to the newly identified IFN-γ ≃B and C3-related sites. In addition, we identified the NFAT proteins as a component of the inducible complexes that bind to the C3- 3P site. Site-directed mutagenesis and transfection studies demonstrate that calcineurin-inducible transcriptional factors enhance the transcriptional activity of the IFN-γ promoter through the cyclosporin-sensitive C3-3P site, whereas NF-≃B proteins functionally interact with the C3-related sites. In addition, when located downstream to the β-galactosidase gene driven by the IFN-γ promoter, the intronic C3 site worked in concert with both the IFN-γ κB and the C3-3P site to enhance gene transcription. These results demonstrate that the coordinate activities of NFAT and NF-≃B proteins are involved in the molecular mechanisms controlling IFN-γ gene transcription.

Original languageEnglish
Pages (from-to)30412-30420
Number of pages9
JournalJournal of Biological Chemistry
Volume272
Issue number48
DOIs
Publication statusPublished - Nov 28 1997

ASJC Scopus subject areas

  • Biochemistry

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    Sica, A., Dorman, L., Viggiano, V., Cippitelli, M., Ghosh, P., Rice, N., & Young, H. A. (1997). Interaction of NF-κB and NFAT with the interferon-γ promoter. Journal of Biological Chemistry, 272(48), 30412-30420. https://doi.org/10.1074/jbc.272.48.30412