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Originally published In Press as doi:10.1074/jbc.M111302200 on March 21, 2002

J. Biol. Chem., Vol. 277, Issue 22, 19408-19417, May 31, 2002
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Isolation and Characterization of a Human STAT1 Gene Regulatory Element
INDUCIBILITY BY INTERFERON (IFN) TYPES I AND II AND ROLE OF IFN REGULATORY FACTOR-1*

Lee H. WongDagger , Helena SimDagger , Moitreyee Chatterjee-Kishore§, Irene HatzinisiriouDagger , Rodney J. DevenishDagger , George Stark§, and Stephen J. RalphDagger

From the Dagger  Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3168, Australia and the § Department of Molecular Biology, Cleveland Clinic Foundation, Cleveland, Ohio 44195

The transcription factor STAT1 plays a pivotal role in signal transduction of type I and II interferons (IFNs). STAT1 activation leads to changes in expression of key regulatory genes encoding caspases and cell cycle inhibitors. Deficient STAT1 expression in human cancer cells and virally mediated inhibition of STAT1 function have been associated with cellular resistance to IFNs and mycobacterial infection in humans. Thus, given the relative importance of STAT1, we isolated and characterized a human STAT1 intronic enhancer region displaying IFN-regulated activity. Functional analyses by transient expression identified a repressor region and type I and II IFN-inducible elements within the STAT1 enhancer sequence. A candidate IRF-E/GAS/IRF-E (IGI) sequence containing GAAANN nucleotide repeats was shown by gel shift assay to bind to IFN regulatory factor-1 (IRF-1), but not to IFN-stimulated gene factor-3 (ISGF-3) or STAT1-3. An additional larger IGI-binding complex containing IRF-1 was identified. Mutation of the GAAANN repeats within the IGI DNA element eliminated IRF-1 binding and the IFN-regulated activity of the STAT1 intronic enhancer region. Transfection of the IFN-resistant MM96 cell line to express increased levels of IRF-1 protein also elevated STAT1, STAT2, and p48/IRF-9 expression and enhanced cellular responsiveness to IFN-beta . Reciprocating regulation between IRF-1 and STAT1 genes and encoded proteins indicates that an intracellular amplifier circuit exists controlling cellular responsiveness to the IFNs.

* 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.

The nucleotide sequences reported in this paper have been submitted to the GenBankTM/EBI Data Bank under accession numbers AF182310-AF182313.

To whom correspondence should be addressed. Tel.: 61-3-9905-37811; Fax: 61-3-9905-4699; E-mail: steve.ralph@med.monash.edu.au.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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