| 崔巍,尹玲玲,董凌月,李莉,刘志成,李延玲,安威,刘晓丹,杨川杰,周平坤,丁库克.早期反应基因Ier5启动子区辐射敏感的转录因子结合位点研究[J].中华放射医学与防护杂志,2012,32(1):15-19 |
| 早期反应基因Ier5启动子区辐射敏感的转录因子结合位点研究 |
| Study on the binding sites of radiosensitivity associated transcription factor in the promoter region of Ier5 gene |
| 投稿时间：2011-01-12 |
| DOI：10.3760/cma.j.issn.0254-5098.2012.01.004 |
| 中文关键词: 早期反应基因5 辐射敏感性 转录 |
| 英文关键词:Immediate early response gene 5 Radiation sensitivity Transcription |
| 基金项目:国家高科技研究发展计划(863) (2004AA221160);国家自然科学基金(30770533;31170806);北京市自然科学基金(7092013);北京市教育委员会科技发展基金(Km200710025007) |
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| 中文摘要: |
| 目的 探讨辐射诱导早期反应基因5(Ier5)的转录调控机制。方法 运用缺失体构建、定点突变、电泳迁移率实验(EMSA)及染色质免疫共沉淀(ChIP)技术,确定宫颈癌HeLa细胞中Ier5基因启动子区域、转录因子及其结合位点。结果 发现Ier5基因的启动子区可能在缺失体Ier5-8所包含的范围(-408~-238 bp)内。Ier5基因具有2个转录因子,分别为GCF、NFI,其中GCF的结合位点在Ier5基因启动子-388~-382 bp和-274~-270 bp处,NFI的结合位点在Ier5基因启动子-362~-357 bp处。GCF抑制Ier5基因的表达,抑制作用随γ射线吸收剂量的增加而减弱,NFI促进Ier5基因的表达。结论 Ier5基因启动子最可能的区域为-408~-238 bp,该区域存在2个辐射敏感的转录因子GCF和NFI的结合位点。GCF对Ier5基因的顺式作用元件起负调节作用,而NFI起正调节作用。 |
| 英文摘要: |
| Objective To clarify the mechanism of immediate early response gene 5 (Ier5) transcription induced by radiation. Methods Deletant construction, site-specific mutagenesis, electrophoretic mobility shift assay(EMSA)and chromatin immunoprecipitation (ChIP) were used to forecast the promoter region, binding sites and transcription factors of Ier5 gene in HeLa cells. Results The promoter region of Ier5 gene might be in the region of Ier5-8 deletant (-408--238 bp). The Ier5 gene had two transcription factors of GCF and NFI, and GCF had two binding sites located in the region of -388--382 bp and -274--270 bp of Ier5 promoter. The binding site of NFI was located in -362--357 bp of Ier5 promoter. GCF could inhibit the expression of Ier5 gene and this inhibition was diminished when the radiation dose increased. In contrast, NFI increased the expression of Ier5. Conclusions The most possible region of Ier5 promoter is from -408 to -238 bp which has two binding sites for the radiosensitivity transcription factors of GCF and NFI that could negatively and positively regulate the expression of Ier5 respectively. |
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