Stimulus-Specific Assembly of Enhancer Complexes on the Tumor Necrosis Factor Alpha Gene Promoter

doi:10.1128/MCB.20.6.2239-2247.2000

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Molecular and Cellular Biology, March 2000, p. 2239-2247, Vol. 20, No. 6
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Stimulus-Specific Assembly of Enhancer Complexes on the Tumor Necrosis Factor Alpha Gene Promoter

James V. Falvo,¹ Adele M. Uglialoro,² Brigitta M. N. Brinkman,² Menie Merika,³ Bhavin S. Parekh,¹ Eunice Y. Tsai,² Hadley C. King,² Anthony D. Morielli,¹ Ernest G. Peralta,¹^, Tom Maniatis,¹ Dimitris Thanos,³ and Anne E. Goldfeld²^,^*

Center for Blood Research and Harvard Medical School, Boston, Massachusetts 02115²; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138¹; Department of Biochemistry and Biophysics, Columbia University, New York, New York 10068³

Received 26 October 1999/Returned for modification 29 November 1999/Accepted 6 December 1999

The human tumor necrosis factor alpha (TNF- $alpha$ ) gene is rapidly activated in response to multiple signals of stress and inflammation.We have identified transcription factors present in the TNF- $alpha$ enhancer complex in vivo following ionophore stimulation (ATF-2/Junand NFAT) and virus infection (ATF-2/Jun, NFAT, and Sp1), demonstratinga novel role for NFAT and Sp1 in virus induction of gene expression.We show that virus infection results in calcium flux and calcineurin-dependentNFAT dephosphorylation; however, relatively lower levels of NFATare present in the nucleus following virus infection as comparedto ionophore stimulation. Strikingly, Sp1 functionally synergizeswith NFAT and ATF-2/c-jun in the activation of TNF- $alpha$ gene transcriptionand selectively associates with the TNF- $alpha$ promoter upon virusinfection but not upon ionophore stimulation in vivo. We concludethat the specificity of TNF- $alpha$ transcriptional activation is achievedthrough the assembly of stimulus-specific enhancer complexes andthrough synergistic interactions among the distinct activatorswithin these enhancercomplexes.

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

Dedicated to the memory of Mauricio X. Zuber and Ernest G.Peralta.

Deceased 17 May1999.

Molecular and Cellular Biology, March 2000, p. 2239-2247, Vol. 20, No. 6
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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