An NF kappa B-like factor is essential but not sufficient for cytokine induction of endothelial leukocyte adhesion molecule 1 (ELAM-1) gene transcription.
The endothelial leukocyte adhesion molecule 1 (ELAM-1) is transiently expressed specifically on the surface of cytokine-induced endothelial cells. We demonstrate that the transient expression of the protein is paralleled by an increase and decrease in transcription of the ELAM-1 gene. To identify the cis-acting transcription control regions within the ELAM-1 gene that are responsible for this cytokine-induced expression, we isolated and analyzed an ELAM-1 genomic clone containing sequences upstream of the transcription start site. We constructed a series of ELAM-1 deletion mutants linked to a reporter gene and analyzed their expression in both endothelial and non-endothelial cells. Results show that a fragment of 233 bp upstream of the transcription start site is sufficient to confer cytokine inducibility upon the reporter gene in both endothelial and non-endothelial cells. Further analysis defined two elements within this region that are involved in the cytokine inducibility of the ELAM-1 gene. One element lies within the -233 to -117 region, the other element represents an NF kappa B consensus binding site between nucleotides -94 to -85. Gel shift analysis reveals increased binding of an NF kappa B-like factor to this consensus sequence in extracts prepared from IL-1-induced endothelial cells. The results suggest that cytokine induction of ELAM-1 gene transcription is imparted by a combination of positive factors, one being an NF kappa B-like transcription factor, interacting with cis-acting elements within the enhancer/promoter of the gene.
Whelan
J
J
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
Ghersa
P
P
Hooft van Huijsduijnen
R
R
Gray
J
J
Chandra
G
G
Talabot
F
F
DeLamarter
J F
JF
D001483:Base Sequence; D002448:Cell Adhesion / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007375:Interleukin-1 / Q000502:physiology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016328:NF-kappa B / Q000502:physiology*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic / Q000502:physiology*
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007375:Interleukin-1; D016328:NF-kappa B; D004247:DNA