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(Received for publication, May 9, 1995; and in revised form, June 7, 1995 ) We investigated the mechanisms by which H
Volume 270,
Number 32,
Issue of August 11, pp. 18966-18974, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
O
and Tumor Necrosis Factor-
Activate Intercellular Adhesion
Molecule 1 (ICAM-1) Gene Transcription through Distinct cis-Regulatory Elements within the ICAM-1 Promoter
O increases intercellular adhesion molecule 1 (ICAM-1; CD54)
expression in endothelial cells. The HO-induced
increase in ICAM-1 mRNA was inhibited by actinomycin D, by the
antioxidant N-acetylcysteine, and by 3-aminobenzamide (which
blocks oxidant-induced AP-1 activity), but not by pyrrolidine
dithiocarbamate (which blocks oxidant-induced NF-
B activity).
Nuclear run-on and transient transfections of ICAM-1 promoter
constructs indicated that HO stimulated ICAM-1
gene transcription by activation of a distinct region of the ICAM-1
promoter. The HO-responsive element was
localized to sequences between -981 and -769 (relative to
the start codon). Located within this region are two 16-base pair
repeats, each containing binding sites for the transcription factors
AP-1 and Ets. A similar composite AP-1/Ets element isolated from the
macrophage scavenger receptor gene conferred HO responsiveness to a minimal promoter. Mutation of the 16-base
pair repeats within the ICAM-1 promoter prevented
HO-induced DNA binding activity, and their
deletion abrogated the HO-induced
transcriptional activity. In contrast, TNF
induced ICAM-1
transcription via activation of promoter sequences between -393
and -176, a region with C/EBP and NF-B binding sites. The
results indicate that HO activates ICAM-1
transcription through AP-1/Ets elements within the ICAM-1 promoter,
which are distinct from NF-B-mediated ICAM-1 expression induced by
TNF.
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