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Originally published In Press as doi:10.1074/jbc.M105121200 on July 25, 2001

J. Biol. Chem., Vol. 276, Issue 45, 41629-41637, November 9, 2001
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Recruitment of Multiple Interferon Regulatory Factors and Histone Acetyltransferase to the Transcriptionally Active Interferon A Promoters*

Wei-Chun AuDagger and Paula M. Pitha§

From the Dagger  Oncology Center and § Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21231

Type I interferon (IFN) plays a critical role in the innate immunity against viral infection. Expression of IFNA genes in infected cells is cell type-dependent and is regulated at the transcriptional level. The present study is focused on the molecular mechanism underlying the differential expression of human IFNA1 and A2 genes. Two nucleotides, at positions -98 and -81 of IFNA1 and A2 promoter, were pivotal to the differential expression. The DNA pull-down and chromatin precipitation assays have shown that nuclear interferon regulatory factor (IRF)-3 and IRF-7 as well as IRF-1 bind to IFNA1 virus-responsive element (VRE). Interestingly, overexpression of IRF-7 increased the otherwise weak binding of both IRF-3 and IRF-7 to IFNA2 VRE. These data together with the results of two-step chromatin immunoprecipitation strongly suggest that the IRF-3 and IRF-7 bind to IFNA1 promoter as a dimer. Furthermore, binding of IRF-3 and IRF-7 to IFNA VRE is associated with the presence of acetylated histone H3, suggesting that histone acetyltransferase(s) is tethered together with virus-activated IRF-3 and IRF-7 to the IFNA1 promoter. In addition, the constitutively active IRF-3 (5D) and IRF-7 (2D) mutants activate the endogenous IFNA genes in uninfected cells; however, the expression profile of IFNA is not identical to that induced by viral infection.

* This work was supported by National Institutes of Health Grant AI 19737-18 and Bridge Award AI 48081 (to P. M. P.).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.

To whom correspondence should be addressed: Johns Hopkins University, Cancer Research Building, 1650 Orleans St., Baltimore, MD 21231. E-mail: parowe@jhmi.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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