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Department of Biochemistry and Biotechnology Institute, Trinity College, Dublin, Ireland
The adhesion molecule CD44 is a multifunctional, ubiquitously
expressed glycoprotein that participates in the process of leukocyte
recruitment to sites of inflammation and to their migration through
lymphatic tissues. In this study, we have investigated the effect of
the proinflammatory cytokine IL-1
on CD44 gene expression in the
human immortalized endothelial cell line ECV304. Immunoblotting of cell
extracts showed constitutive expression of a 85-kDa protein
corresponding to the standard form of CD44, which was potently
up-regulated following IL-1 treatment. Furthermore, IL-1 induced
expression of v3- and v6-containing isoforms of CD44, which migrated at
110 and 140–180 kDa, respectively. The effect of IL-1 on CD44
standard, v3- and v6-containing isoforms was dose and time dependent
and was inhibited in the presence of IL-1 receptor antagonist. To
elucidate the molecular mechanisms regulating CD44 expression in
response to IL-1, we investigated the effect of IL-1 on CD44 mRNA
expression. Reverse-transcriptase PCR and Northern analysis
demonstrated an increase in CD44 mRNA expression indicating a
transcriptional mechanism of control by IL-1. Furthermore, IL-1
increased expression of a reporter gene under the control of the CD44
promoter (up to -1.75 kb). The effect of IL-1 was critically
dependent on the site spanning -151 to -701 of the promoter. This
effect required the presence of an Egr-1 motif at position -301 within
the CD44 promoter since mutation of this site abolished responsiveness.
IL-1 also induced Egr-1 expression in these cells. These studies
therefore identify Egr-1 as a critical transcription factor involved in
CD44 induction by IL-1.
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