title: Cloning and characterizing of the murine IRF-3 gene promoter region.
abstract: 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.
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China.|Department of Laboratory Medicine, The First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, Jiangsu Province, China.
authors.forename: Hua-Guo
authors.initials: HG
authors.lastname: Xu
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China.
authors.forename: Lifei
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Jin
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ren
authors.affiliation: Department of Pediatrics, The First Affiliated Hospital, Nanjing Medical University, 300 Guang Zhou Road, Nanjing, 210029, Jiangsu Province, China. gpzhou2003@126.com.
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Zhou
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D007109:Immunity / Q000235:genetics; D007249:Inflammation / Q000235:genetics / Q000276:immunology*; D050838:Interferon Regulatory Factor-3 / Q000235:genetics*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D041681:NIH 3T3 Cells; D015221:Potassium Channels / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D015533:Transcriptional Activation; D050986:YY1 Transcription Factor / Q000378:metabolism*
chemical_list: C488358:Erg3 protein, mouse; D050838:Interferon Regulatory Factor-3; D015221:Potassium Channels; D034741:RNA, Small Interfering; D050986:YY1 Transcription Factor; C494699:Yy1 protein, mouse
keywords: Egr2; Murine interferon regulatory factor 3 (mIRF-3); Promoter; Transcriptional activity; YY1