en
Scientific article
English

Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages

Published inThe Journal of experimental medicine, vol. 171, no. 1, p. 35-47
Publication date1990
Abstract

We have explored the cis-acting elements necessary for the LPS-mediated activation of the mouse TNF-alpha promoter by transfecting a set of 5' deletion mutants linked to the CAT reporter gene into primary bone marrow-derived macrophages. A major drop in inducibility by LPS was seen upon deletion of a region mapping between nt -655 and nt -451. Gel retardation assays revealed that LPS induced the appearance in this region of several specific DNA-protein complexes mapping to sequence motifs with strong homology to the kappa B enhancer. Constructs containing two or more copies of one of the kappa B enhancer motifs linked to a heterologous promoter were inducible by LPS. Additional deletion of a region between nt -301 and nt -241, which contains a MHC class II-like "Y box" and formed a Y box-specific complex with a protein whose concentration was increased by LPS, caused a nearly complete loss of inducibility by LPS. We speculate that NF-kappa B and/or related proteins are involved in the LPS-induced transcriptional activation of the TNF-alpha gene, and that factors interacting with the Y box can additionally modulate the activity of the gene in macrophages.

Keywords
  • Animals
  • B-Lymphocytes/immunology
  • Base Sequence
  • Cells, Cultured
  • Enhancer Elements, Genetic
  • Gene Expression Regulation
  • Genes/drug effects
  • Genes, Immunoglobulin
  • Humans
  • Immunoglobulin kappa-Chains/genetics
  • Lipopolysaccharides/pharmacology
  • Macrophages/drug effects/immunology
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Plasmids
  • Promoter Regions, Genetic
  • Restriction Mapping
  • Sequence Homology, Nucleic Acid
  • Transcription, Genetic/drug effects
  • Tumor Necrosis Factor-alpha/genetics
Citation (ISO format)
SHAKHOV, A N et al. Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages. In: The Journal of experimental medicine, 1990, vol. 171, n° 1, p. 35–47. doi: 10.1084/jem.171.1.35
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Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0022-1007
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