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Molecular and Cellular Biology, August 2000, p. 6084-6094, Vol. 20, No. 16
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Lipopolysaccharide-Specific Enhancer Complex Involving Ets, Elk-1, Sp1, and CREB Binding Protein and p300 Is Recruited to the Tumor Necrosis Factor Alpha Promoter In Vivo

Eunice Y. Tsai,1 James V. Falvo,2 Alla V. Tsytsykova,1 Amy K. Barczak,1 Andreas M. Reimold,3 Laurie H. Glimcher,3 Matthew J. Fenton,4 David C. Gordon,1 Ian F. Dunn,1 and Anne E. Goldfeld1,*

The Center for Blood Research and Harvard Medical School1 and The Harvard School of Public Health,3 Boston, Massachusetts 02115; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 021382; and Pulmonary Center, Boston University School of Medicine, Boston, Massachusetts 021184

Received 10 May 2000/Accepted 19 May 2000

The tumor necrosis factor alpha (TNF-alpha ) gene is rapidly activated by lipopolysaccharide (LPS). Here, we show that extracellular signal-regulated kinase (ERK) kinase activity but not calcineurin phosphatase activity is required for LPS-stimulated TNF-alpha gene expression. In LPS-stimulated macrophages, the ERK substrates Ets and Elk-1 bind to the TNF-alpha promoter in vivo. Strikingly, Ets and Elk-1 bind to two TNF-alpha nuclear factor of activated T cells (NFAT)-binding sites, which are required for calcineurin and NFAT-dependent TNF-alpha gene expression in lymphocytes. The transcription factors ATF-2, c-jun, Egr-1, and Sp1 are also inducibly recruited to the TNF-alpha promoter in vivo, and the binding sites for each of these activators are required for LPS-stimulated TNF-alpha gene expression. Furthermore, assembly of the LPS-stimulated TNF-alpha enhancer complex is dependent upon the coactivator proteins CREB binding protein and p300. The finding that a distinct set of transcription factors associates with a fixed set of binding sites on the TNF-alpha promoter in response to LPS stimulation lends new insights into the mechanisms by which complex patterns of gene regulation are achieved.

* Corresponding author. Mailing address: The Center for Blood Research and Harvard Medical School, 800 Huntington Ave., Boston, MA 02115. Phone: (617) 278-3351. Fax: (617) 278-3454. E-mail: goldfeld{at}cbr.med.harvard.edu.

Molecular and Cellular Biology, August 2000, p. 6084-6094, Vol. 20, No. 16
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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