title: Core promoter analysis of porcine Six1 gene and its regulation of the promoter activity by CpG methylation.
abstract: Six1, an evolutionary conserved transcription factor, has been shown to play an important role in organogenesis and diseases. However, no reports were shown to investigate its transcriptional regulatory mechanisms. In the present study, we first identified porcine Six1 gene core promoter region (+170/-360) using luciferase reporter assay system and found that promoter activities were significantly higher in the mouse myoblast C2C12 cells than that in the mouse fibroblast C3H10T1/2 cells, implying that Six1 promoter could possess muscle-specific characteristics. Moreover, our results showed that promoter activities of Six1 were decreased as induction of differentiation of C2C12 cells, which was accompanied by the down-regulation of mRNA expression of Six1 gene. In addition, we found that the DNA methylation of Six1 promoters in vitro obviously influences the promoter activities and the DNA methylation level of Six1 promoter core region was negatively correlated to Six1 gene expression in vivo. Taken together, we preliminarily clarified transcriptional regulatory mechanisms of Six1 gene, which should be useful for investigating its subtle transcriptional regulatory mechanisms in the future. On the other hand, based on Six1 involved in tumorigenesis, our data also provide a genetic foundation to control the generation of diseases via pursuing Six1 as therapeutic target gene.
authors: Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: wuwangjun2009@hotmail.com.
authors: Wangjun
authors: W
authors: Wu
authors: Zhuqing
authors: Z
authors: Ren
authors: Honglin
authors: H
authors: Liu
authors: Linjie
authors: L
authors: Wang
authors: Ruihua
authors: R
authors: Huang
authors: Jie
authors: J
authors: Chen
authors: Lin
authors: L
authors: Zhang
authors: Pinghua
authors: P
authors: Li
authors: Yuanzhu
authors: Y
authors: Xiong
mesh_terms: D000818:Animals; D002460:Cell Line; D018899:CpG Islands; D019175:DNA Methylation; D015536:Down-Regulation; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D018398:Homeodomain Proteins; D012333:RNA, Messenger; D014157:Transcription Factors
keywords: DMEM; DNA methylation; Drosophila sine oculis; Dulbecco's modified Eagle's medium; Eya1; FBS; HD; MyoD; Promoter; RACE technology; Rapid-Amplification of cDNA Ends technology; SD; Six domain; Six1; So; TGF-β signaling; TSS; Transcriptional regulation; days post-conception; dpc; eyes absent homolog 1; fetal bovine serum; homeodomain; myogenic differentiation factor; sine oculis homeobox 1; transcription start site; transforming growth factor-beta signaling