Identification of Distinct Regions of 5′ Flanking DNA That Mediate Constitutive, IFN-γ, STAT1, and TGF-β-Regulated Expression of the Class II Transactivator Gene

Abstract

Abstract Class II transactivator (CIITA) is a master regulator required for constitutive and IFN-γ-inducible expression of class II MHC genes. Although the role of CIITA is greatly appreciated, the mechanisms underlying constitutive and IFN-γ-induced expression of CIITA are not understood. The study of CIITA induction is extremely important, but has been fraught with difficulty. This study describes for the first time a large (7-kb) fragment of 5′ flanking sequences that mediates the B cell-specific, IFN-γ-induced, and TGF-β-suppressed expression of CIITA. This pattern of expression matches the authentic expression of the endogenous gene. Within the 7-kb fragment, sequences that lie between nucleotides −545 and −113 relative to the transcriptional start site are critical for constitutive promoter expression in B cells. In contrast, inducible activation of CIITA by IFN-γ requires sequences contained in an additional 4 kb of upstream DNA. This region mediates an IFN-γ response when linked to either the endogenous CIITA promoter or a heterologous promoter. A role for STAT1 in regulation of the CIITA promoter is shown by the rescue of IFN-γ induction by expression of STAT1 in STAT1-defective U3A cells. TGF-β significantly inhibits IFN-γ-mediated induction of the CIITA promoter in 2fTGH fibroblasts, which indicates that the promoter is a target for TGF-β. This inhibition is achieved by suppression of the basal promoter. This study provides a focal point for understanding the mechanism of B cell-specific, IFN-γ-induced, and TGF-β-suppressed expression of CIITA.