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J Biol Chem, Vol. 275, Issue 16, 11750-11757, April 21, 2000
From the The cyclooxygenase (COX)-2 enzyme is responsible
for increased prostaglandin formation in inflammatory states and is the
major target of nonsteroidal anti-inflammatory drugs. Normally COX-2 expression is tightly regulated, however, constitutive overexpression plays a key role in colon carcinogenesis. To understand the mechanisms controlling COX-2 expression, we examined the ability of the
3'-untranslated region of the COX-2 mRNA to regulate
post-transcriptional events. When fused to a reporter gene, the
3'-untranslated region mediated rapid mRNA decay
(t1/2 = 30 min), which was comparable to endogenous
COX-2 mRNA turnover in serum-induced fibroblasts treated with
actinomycin D or dexamethasone. Deletion analysis demonstrated that a
conserved 116-nucleotide AU-rich sequence element (ARE) mediated
mRNA degradation. In transiently transfected cells, this region
inhibited protein synthesis approximately 3-fold. However, this
inhibition did not occur through changes in mRNA stability since
mRNA half-life and steady-state mRNA levels were unchanged. RNA
mobility shift assays demonstrated a complex of cytoplasmic proteins
that bound specifically to the ARE, and UV cross-linking studies
identified proteins ranging from 90 to 35 kDa. Fractionation of the
cytosol showed differential association of ARE-binding proteins to
polysomes and S130 fractions. We propose that these factors influence
expression at a post-transcriptional step and, if dysregulated, may
increase COX-2 protein as detected in colon cancer.
Post-transcriptional Control of Cyclooxygenase-2 Gene
Expression
THE ROLE OF THE 3'-UNTRANSLATED REGION*
§,§
Department of Oncological Sciences, Eccles
Program in Human Molecular Biology and Genetics, and the Huntsman
Cancer Institute, University of Utah,
Salt Lake City, Utah 84112
*
This work was supported by American Heart Association
Scientist Development Grant 9930102N (to D. A. D.) and
National Institutes of Health (NCI) Grant CA73992. The core facilities
at the Huntsman Cancer Institute (DNA sequencing and DNA/peptide
synthesis) are supported by Cancer Center Support Grant CA42014 from
the National Institutes of Health (NCI).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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