DNA-methylation of the E-selectin promoter represses NF-kappa B transactivation

Biochem Biophys Res Commun. 1993 Jul 15;194(1):215-21. doi: 10.1006/bbrc.1993.1806.

Abstract

E-selectin is an adhesion molecule transiently and specifically expressed on endothelial cells upon stimulation with cytokines. We wished to determine whether methylation could play a role in cell-type specific expression of this gene. We found that the E-selectin promoter in cultured endothelial cells is under-methylated in comparison with non-expressing HeLa cells. Plasmid constructs carrying a reporter driven by the E-selectin promoter and methylated in vitro are no longer transcribed in either an in vitro transcription system or in transiently transfected cells. We identified the NF-kappa B site in the promoter as the likely target for this methylation-mediated repression by testing a minimal promoter carrying only this and an associated element. We conclude that methylation is likely to play a role in blocking E-selectin expression in non-endothelial cells.

MeSH terms

  • Base Sequence
  • Cell Adhesion Molecules / genetics*
  • Cells, Cultured
  • Chromatography, Affinity
  • DNA / isolation & purification
  • DNA / metabolism*
  • E-Selectin
  • Endothelium, Vascular / metabolism*
  • HeLa Cells
  • Humans
  • Membrane Glycoproteins / genetics*
  • Methylation
  • Molecular Sequence Data
  • NF-kappa B / antagonists & inhibitors*
  • Oligodeoxyribonucleotides
  • Plasmids
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic*
  • Transcription, Genetic
  • Transcriptional Activation
  • Transfection
  • Umbilical Veins

Substances

  • Cell Adhesion Molecules
  • E-Selectin
  • Membrane Glycoproteins
  • NF-kappa B
  • Oligodeoxyribonucleotides
  • DNA