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The Journal of Immunology, 2003, 171: 4187-4194.
Copyright © 2003 by The American Association of Immunologists

IFN-{gamma}-Induced MHC Class II Expression: Transactivation of Class II Transactivator Promoter IV by IFN Regulatory Factor-1 is Regulated by Protein Kinase C-{alpha} 1

Mélanie Giroux*, Manuel Schmidt{dagger} and Albert Descoteaux2,*

* Institut National de la Recherche Scientifique–Institut Armand-Frappier, Université du Québec, Laval, Québec, Canada; and {dagger} Mologen GmbH, Berlin, Germany

Previous studies based on pharmacological evidence suggested a requirement for protein kinase C (PKC) activity in the regulation of IFN-{gamma}-induced MHC class II (MHC-II) expression. In the present study, we investigated the molecular mechanisms by which PKC-{alpha} modulates IFN-{gamma}-induced MHC-II expression in the mouse macrophage cell line RAW 264.7. Overexpression of a dominant-negative (DN) mutant of PKC-{alpha} inhibited the expression of IFN-{gamma}-induced MHC-II but had no effect on IFN-{gamma}-induced STAT1 nuclear translocation and DNA binding activity, as well as on the expression of inducible NO synthase, IFN consensus sequence binding protein, MHC class I, IFN regulatory factor (IRF)-1, and IFN-{gamma}-inducible protein-10. Further analysis showed that IFN-{gamma}-induced expression of the MHC class II transactivator (CIITA), a transcriptional coactivator essential for MHC-II expression, was inhibited in DN PKC-{alpha}-overexpressing cells. Studies with reporter constructs containing the promoter IV region of CIITA revealed that overexpression of a constitutively active mutant of PKC-{alpha} enhanced IRF-1, but not IRF-2, transcriptional activity. Furthermore, characterization of IRF-1 from both normal and DN PKC-{alpha}-overexpressing cells revealed differences in IRF-1 posttranslational modifications. Collectively, our data suggest a novel regulatory mechanism for IFN-{gamma}-induced MHC-II expression, whereby PKC regulates CIITA expression by selectively modulating the transcriptional activity of IRF-1.




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