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J. Biol. Chem., Vol. 275, Issue 42, 32688-32693, October 20, 2000
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From the Department of Cancer Cell Biology, Harvard School of
Public Health, Boston, Massachusetts 02115
Expression of heme oxygenase-1 (HO-1) in
mammalian cells contributes to resistance to various types of free
radical damage. Nitric oxide (NO) induces HO-1 in many cell types, but
the specific contribution of transcriptional or post-transcriptional
effects to this induction have remained unresolved. Here we show that the extent of HO-1 mRNA expression in IMR-90 and HeLa cells depends on the rate of NO delivery, and that the induction occurs more slowly
in HeLa than in human fibroblast (IMR-90) cells. We used a specific NO
scavenger
(2-(4-carboxylphenyl)-4,4,5,5-tetramethylimidazolin-1-oxyl 3-oxide)
that completely prevented the inducible expression of HO-1 by NO,
pointing to direct signaling action of NO in this induction. By
inhibiting transcription during the NO exposure, we have confirmed that
NO treatment activates a mechanism that stabilizes HO-1 mRNA. The
increase in the HO-1 mRNA half-life in IMR-90 cells was directly
correlated with increasing rates of NO release. We also show here that
the stabilization of the HO-1 message does not require de
novo protein synthesis. Collectively, these results show that
stabilization of HO-1 mRNA can be finely tuned to the NO exposure,
and that the effect in human fibroblasts is mediated by a pre-existing protein.
Nitric Oxide-inducible Expression of Heme Oxygenase-1 in
Human Cells
TRANSLATION-INDEPENDENT STABILIZATION OF THE mRNA AND
EVIDENCE FOR DIRECT ACTION OF NITRIC OXIDE*
and
*
This work was supported in part by a fellowship (to C. B.) from the Association pour la Recherche contre le Cancer and by National Institutes of Health Grant CA82737 (to B. D.).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.
Current address: CNRS, Institut de Chimie des Substances
Naturelles, 91190 Gif-sur-Yvette, France.
§
To whom correspondence should be addressed. Tel.: 617-432-2286;
Fax: 617-432-0377; E-mail: bdemple@hsph.harvard.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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