Cloning and characterizing of the murine IRF-3 gene promoter region

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• 作者：Hua-Guo Xu ; Lifei Liu ; Shan Gao ; Rui Jin ; Wei Ren ; Guo-Ping Zhou
• 刊名：Immunologic Research
• 出版年：2016
• 出版时间：August 2016
• 年：2016
• 卷：64
• 期：4
• 页码：969-977
• 全文大小：1,252 KB
• 刊物主题：Allergology; Immunology; Medicine/Public Health, general; Internal Medicine;
• 出版者：Springer US
• ISSN：1559-0755
• 卷排序：64

文摘

The interferon regulatory factor 3 (IRF-3) plays essential roles in inflammation and immune response. Here, we cloned the nucleotide sequence of the 5′-flanking region of the murine IRF-3 gene (mIRF-3) and characterized the molecular mechanisms controlling the mIRF-3 transcriptional activity in NIH3T3 cells. Analyses of a series of 5′ deletion constructs demonstrated that a 301 bp region (−255/+46) of the mIRF-3 gene is sufficient for full promoter activity. This region contains IK1, Egr2, Cmyb, E2F1 and YY1 putative transcription factor binding sites. Mutation of Egr2 or YY1 site led to 52–68 % decrease of the mIRF-3 promoter activity, and double Egr2 and YY1 mutation reduced the promoter activity to 20 % of the wild-type promoter activity. Furthermore, knockingdown of endogenous Egr2 or YY1 by a siRNA strategy markedly inhibited the mIRF-3 promoter activity. Chromatin immunoprecipitation assays showed that Egr2 and YY1 interact with the mIRF-3 promoter in vivo. These results suggested that the basal promoter activity of the mIRF-3 gene is regulated by transcription factors Egr2 and YY1 in NIH3T3 cells.KeywordsMurine interferon regulatory factor 3 (mIRF-3)Transcriptional activityPromoterYY1Egr2