TNF Regulates the in Vivo Occupancy of Both Distal and Proximal Regulatory Regions of the MCP-1/JE Gene

Immunity. 1996 May;4(5):455-69. doi: 10.1016/s1074-7613(00)80412-4.


In vivo genomic footprinting (IVGF) was used to examine regulatory site occupancy during the activation of the murine inflammatory response gene MCP-1/JE by TNF. In response to TNF, both promoter distal and proximal regulatory regions became occupied in vivo. EMSA analysis showed that while some of the factors involved in expression, including NF-kappa B, were translocated to the nucleus following TNF treatment, others were already present and able to bind DNA in vitro. Protein kinase inhibitor studies showed that protein phosphorylation was required for TNF activation but not factor assembly. These studies provide evidence for a multistep model of TNF-mediated gene regulation involving chromatin accessibility, transcription factor complex assembly, and protein phosphorylation.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Animals
  • Base Sequence
  • Binding, Competitive / drug effects
  • Cell Nucleus / genetics
  • Chemokine CCL2 / genetics*
  • Codon / drug effects
  • Cycloheximide / pharmacology
  • Gene Expression Regulation / drug effects*
  • Gene Expression Regulation / immunology
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • NF-kappa B / genetics
  • Proline / analogs & derivatives
  • Proline / pharmacology
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / immunology
  • Protein Kinases / adverse effects
  • Regulatory Sequences, Nucleic Acid / drug effects*
  • Regulatory Sequences, Nucleic Acid / immunology
  • Thiocarbamates / pharmacology
  • Transcription Factor AP-1 / genetics
  • Transcription Factors / drug effects
  • Transcription Factors / genetics
  • Tumor Necrosis Factor-alpha / pharmacology*


  • Chemokine CCL2
  • Codon
  • NF-kappa B
  • Thiocarbamates
  • Transcription Factor AP-1
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • prolinedithiocarbamate
  • Cycloheximide
  • Proline
  • Protein Kinases