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

Mol Cell Biol. 2000 Aug;20(16):6084-94. doi: 10.1128/MCB.20.16.6084-6094.2000.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • CREB-Binding Protein
  • Cell Line
  • DNA-Binding Proteins*
  • Enhancer Elements, Genetic
  • Gene Expression Regulation / drug effects
  • Lipopolysaccharides / pharmacology
  • Molecular Sequence Data
  • Nuclear Proteins / genetics*
  • Promoter Regions, Genetic / genetics
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins c-ets
  • Signal Transduction / genetics
  • Sp1 Transcription Factor / genetics*
  • Trans-Activators / genetics*
  • Transcription Factors / genetics*
  • Tumor Necrosis Factor-alpha / genetics*
  • ets-Domain Protein Elk-1


  • DNA-Binding Proteins
  • Lipopolysaccharides
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-ets
  • Sp1 Transcription Factor
  • Trans-Activators
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • ets-Domain Protein Elk-1
  • CREB-Binding Protein