PMID:17159910

Citation	Byun, SJ, Jeon, IS, Lee, H and Kim, TY (2007) IFN-gamma upregulates expression of the mouse complement C1rA gene in keratinocytes via IFN-regulatory factor-1. J. Invest. Dermatol. 127:1187-96
Abstract	We examined the expression of the mouse complement component C1rA (mC1rA) in IFN-gamma-stimulated mouse keratinocytes (Pam 212) and found that it was upregulated. To analyze the mechanism involved, we cloned the 2,150 bp 5'-flanking region of mC1rA by the vectorette-PCR technique, and identified the transcription start site of mC1rA by rapid amplification of complementary DNA ends. Analysis of the 5' sequence revealed putative binding sites for activator protein 1, CCAAT/enhancer binding protein (C/EBP), signal transducer and activator of transcription 1 (STAT-1), IFN-regulatory factor-1 (IRF-1), and others. We detected transcriptional activation dependent on this upstream region in reporter gene assays and Northern blots. To identify the cis-acting regulatory elements involved, we analyzed serial deletion constructs of the promoter using luciferase reporters. The -80 to -19 bp region, which contains a putative IRF-1 binding site, was required for both basal promoter activity and responses to IFN-gamma. The use of site-directed point mutations, electrophoresis mobility shift assays, and supershift assays indicated that the putative IRF-1 binding site was essential for both IFN-gamma-dependent and -independent transcriptional activity of the mC1rA promoter. We conclude that IFN-gamma stimulates mC1rA gene expression via IRF-1 in mouse keratinocytes.
Links	PubMed Online version:10.1038/sj.jid.5700660
Keywords	Animals; Base Sequence; Binding Sites/genetics; Binding Sites/physiology; CCAAT-Enhancer-Binding Proteins/genetics; CCAAT-Enhancer-Binding Proteins/physiology; Cell Line; Complement C1r/genetics; Complement C1r/metabolism; Humans; Interferon Regulatory Factor-1/genetics; Interferon Regulatory Factor-1/physiology; Interferon-gamma/physiology; Keratinocytes/cytology; Keratinocytes/physiology; Mice; Mice, Inbred BALB C; Molecular Sequence Data; STAT1 Transcription Factor/genetics; STAT1 Transcription Factor/physiology; Transcription Factor AP-1/genetics; Transcription Factor AP-1/physiology; Up-Regulation/genetics; Up-Regulation/physiology
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