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Mol Cell Biol, June 1998, p. 3395-3404, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Interaction of Stat6 and NF-kappa B: Direct Association and Synergistic Activation of Interleukin-4-Induced Transcription

Ching-Hung Shen and Janet Stavnezer*

Department of Molecular Genetics and Microbiology and Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, Massachusetts 01655-0122

Received 8 September 1997/Returned for modification 9 October 1997/Accepted 19 March 1998

Signal transducer and activator of transcription 6 (Stat6) and NF-kappa B are widely distributed transcription factors which are induced by different stimuli and bind to distinct DNA sequence motifs. Interleukin-4 (IL-4), which activates Stat6, synergizes with activators of NF-kappa B to induce IL-4-responsive genes, but the molecular mechanism of this synergy is poorly understood. Using glutathione S-transferase pulldown assays and coimmunoprecipitation techniques, we find that NF-kappa B and tyrosine-phosphorylated Stat6 can directly bind each other in vitro and in vivo. An IL-4-inducible reporter gene containing both cognate binding sites in the promoter is synergistically activated in the presence of IL-4 when Stat6 and NF-kappa B proteins are coexpressed in human embryonic kidney 293 (HEK 293) cells. The same IL-4-inducible reporter gene is also synergistically activated by the endogenous Stat6 and NF-kappa B proteins in IL-4-stimulated I.29µ B lymphoma cells. Furthermore, Stat6 and NF-kappa B bind cooperatively to a DNA probe containing both sites, and the presence of a complex formed by their cooperative binding correlates with the synergistic activation of the promoter by Stat6 and NF-kappa B. We conclude that the direct interaction between Stat6 and NF-kappa B may provide a basis for synergistic activation of transcription by IL-4 and activators of NF-kappa B.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, 55 Lake Ave. North, Worcester, MA 01655-0122. Phone: (508) 856-4100. Fax: (508) 856-1789. E-mail: Janet.Stavnezer{at}banyan.ummed.edu.

Mol Cell Biol, June 1998, p. 3395-3404, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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