DNA-methylation of the E-selectin promoter represses NF-kappa B transactivation.
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.
Smith
G M
GM
GLAXO Institute for Molecular Biology, Chemin des Aulx 1228, Plan-les-Ouates Geneva, Switzerland.
Whelan
J
J
Pescini
R
R
Ghersa
P
P
DeLamarter
J F
JF
Hooft van Huijsduijnen
R
R
D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000235:genetics*; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics*; D008745:Methylation; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014471:Umbilical Veins
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D004247:DNA