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Department of Microbiology, Immunology, and Molecular Genetics, and Jonsson Comprehensive Cancer Center, University of California School of Medicine, Los Angeles, CA 90095
NO has been increasingly implicated in control of the
transcriptional machinery and serves as an intracellular second
messenger to modify gene expression. We have demonstrated that NO
up-regulated Fas receptor expression in ovarian carcinoma cell lines,
albeit the mechanism involved is not known. Thus, we hypothesized that
NO, directly or indirectly, may modify the transcriptional machinery
that is responsible for the increased expression of the Fas gene. We
examined the effect of NO on Fas gene expression using a Fas
promoter-driven luciferase reporter system. Transient transfection of
AD10 cells with pGL-3-FasP demonstrated that the IFN-
-dependent NO
generation increases the trans-activation of the Fas
promoter, and this increase was blocked by the NOS inhibitor
(NG-monomethyl-L-arginine), but
could be restored by the addition of the NO donor
S-nitroso-N-acetylpenicillamine.
Systematic deletion of the Fas promoter revealed that the functional
region responsible for the NO-mediated effect was located at the
silencer region, suggesting that NO may be responsible for the
disruption of a repressor mechanism. We demonstrate that NO
up-regulates the expression of the Fas receptor on AD10 cells via the
specific inactivation of the transcription repressor yin-yang 1 DNA
binding activity to the silencer region of the Fas promoter. These
findings reveal a new mechanism of NO-mediated gene regulation by
interfering with a repressor transcription factor at the silencer
region of the Fas promoter.
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