Functional analysis of multiple transcription factor sites in a regulatory element of human epsilon-globin gene
文献类型：期刊论文
| 作者 | Hou, CH; Huang, J; Qian, RL |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2004 |
| 卷号 | 36期号:10页码:673-680 |
| 关键词 | epsilon-globin gene regulatory element NF-kappa B GATA |
| 通讯作者 | Qian, RL (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol,Grad Sch, Shanghai 200031, Peoples R China.,rlqian@sibs.ac.cn |
| 英文摘要 | The developmental control of the human epsilon-globin gene expression is mediated by transcriptional regulatory elements in the 5' flanking DNA of this gene. A previously identified negative regulatory element (-3028 to -2902 bp, termed epsilon-NRAII) was analyzed and one putative NF-kappaB site and two GATA sites locate at -3004 bp, -2975 bp and -2948 bp were characterized. Electrophoresis mobility shift assay (EMSA) showed that the putative NF-kappaB site was specifically bound by nuclear proteins of K562 cells. Data obtained from transient transfection showed that the expression of reporter gene could be upregulated about 50% or 100% respectively when epsilon-NRAII was inserted upstream of the SV40 promoter or epsilon-globin gene proximal promoter (-177 bp to +1 bp), suggesting that epsilon-NRAII might not be a classic silencer. Mutation in the putative NF-kappaB site or in the GATA site (at -2975 bp) slightly reduced the expression of reporter gene driven by SV40 promoter or epsilon-globin gene proximal promoter. However, the mutation of GATA site at -2948 bp remarkably reduced the reporter gene activity driven by SV40 promoter, but not by epsilon-globin gene proximal promoter. Further mutation analysis showed that the negative effect of mutation in GATA site at -2948 bp on SV40 promoter was not affected by the mutation of the putative NF-kappaB site, whereas it could be abolished by the mutation of GATA site at -2975 bp. Furthermore, the mutation of both GATA sites could synergistically reduce the reporter gene activity driven by epsilon-globin gene proximal promoter. Those results suggested that epsilon-NRAII might function differently on the SV40 promoter and epsilon-globin gene proximal promoter. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | TRANSGENIC MICE ; DEVELOPMENTAL CONTROL ; SILENCER ; REGION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000224758500005 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2133]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Hou, CH,Huang, J,Qian, RL. Functional analysis of multiple transcription factor sites in a regulatory element of human epsilon-globin gene[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2004,36(10):673-680. |
| APA | Hou, CH,Huang, J,&Qian, RL.(2004).Functional analysis of multiple transcription factor sites in a regulatory element of human epsilon-globin gene.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,36(10),673-680. |
| MLA | Hou, CH,et al."Functional analysis of multiple transcription factor sites in a regulatory element of human epsilon-globin gene".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 36.10(2004):673-680. |
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