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M111302200v1
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Papers In Press, published online ahead of print March 21, 2002
J. Biol. Chem, 10.1074/jbc.M111302200
Submitted on November 27, 2001
Revised on March 13, 2002
Accepted on March 20, 2002

Isolation and characterization of a human STAT1 gene regulatory element. Inducibility by IFN types I and II and role of IRF-1

Lee H. Wong, Helena Sim, Moitreyee Chatterjee-Kishore, Irene Hatzinisiriou, Rodney J. Devenish, George Stark, and Stephen J. Ralph

Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800

Corresponding Author: steve.ralph{at}med.monash.edu.au

The transcription factor STAT1 plays a pivotal role in signal transduction of type I and II 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 viral mediated inhibition of STAT1 function has been associated with cellular resistance to IFNs and mycobacterial infection in humans. Thus, given the relative importance of STAT1, we isolated and characterised 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 IRF-1 but not to ISGF3 or STATs 1-3. 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 IFNbeta . Reciprocating regulation between IRF-1 and STAT1 genes and encoded proteins indicates that an intracellular amplifier circuit exists controlling cellular responsiveness to the IFNs.


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