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The Journal of Immunology, 1998, 161: 4736-4744.
Copyright © 1998 by The American Association of Immunologists

p48/STAT-1{alpha}-Containing Complexes Play a Predominant Role in Induction of IFN-{gamma}-Inducible Protein, 10 kDa (IP-10) by IFN-{gamma} Alone or in Synergy with TNF-{alpha}1

Sarmila Majumder2,*, Lucy Z.-H. Zhou*, Priya Chaturvedi3,*, Gerald Babcock*, Sumer Aras4,* and Richard M. Ransohoff5,*,{dagger}

* Department of Neurosciences, Lerner Research Institute, Cleveland Clinic Foundation, and {dagger} Neurology Department, Mellen Center for Multiple Sclerosis Treatment and Research, Cleveland Clinic Foundation, Cleveland, OH 44195

Human IFN-{gamma}-inducible protein, 10 kDa (hIP-10) and murine IP-10 (mIP-10) genes are induced by IFN-{gamma} alone, and synergistically induced by TNF-{alpha} and IFN-{gamma}. 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-{gamma}. Trans-acting factors mediating the IFN-{gamma} 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-{gamma}, with or without TNF-{alpha}, 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-{gamma}/TNF-{alpha}-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-{alpha}/IFN-{gamma} also required NF{kappa}B binding sites, which were protected in vivo and bound p65 homodimeric NF{kappa}B in vitro. These results documented the essential role of p48 (complexed with STAT-1{alpha}) 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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