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-Containing Complexes Play a Predominant Role in Induction of IFN-
-Inducible Protein, 10 kDa (IP-10) by IFN-
Alone or in Synergy with TNF-
1

*
Department of Neurosciences, Lerner Research Institute, Cleveland Clinic Foundation, and
Neurology Department, Mellen Center for Multiple Sclerosis Treatment and Research, Cleveland Clinic Foundation, Cleveland, OH 44195
Human IFN-
-inducible protein, 10 kDa (hIP-10) and murine IP-10
(mIP-10) genes are induced by IFN-
alone, and synergistically
induced by TNF-
and IFN-
. Upstream regions of the human and
murine genes contain conserved regulatory motifs, including an
IFN-stimulated response element (ISRE), which governs response of the
mIP-10 gene to IFN-
. Trans-acting factors mediating the
IFN-
response via ISRE remain incompletely defined. We examined
ISRE-binding factors in the regulation of the hIP-10 gene. The
requirement of p48 for hIP-10 induction by IFN-
, with or without
TNF-
, was demonstrated using p48-deficient U2A cells. An hIP-10
promoter-reporter mutant (mISRE3) that was relatively deficient for
binding a related factor, IFN regulatory factor-1 (IRF-1) but competent
for binding p48, was induced as well as the wild-type hIP-10 promoter,
supporting the interpretation that p48 played a necessary and
sufficient role in hIP-10 transcription. Genomic in vivo footprinting
revealed IFN-
/TNF-
-inducible binding at the ISRE consistent with
the presence of p48 and associated factors, but not with IRF-1.
Induction of hIP-10 by TNF-
/IFN-
also required NF
B binding
sites, which were protected in vivo and bound p65 homodimeric NF
B in
vitro. These results documented the essential role of p48 (complexed
with STAT-1
) for induction and sustained transcription of the IP-10
gene, strongly suggesting that IRF-1 is not required for IP-10
induction by these inflammatory cytokines.
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