NFY结合位点缺失下调NPCEDRG基因启动子转录活性和效率

发布时间：2018-12-18 21:18

【摘要】：NPCEDRG基因是一个鼻咽癌(nasopharyngeal carcinoma,NPC)相关基因.NPCEDRG基因启动子为TATA-less启动子,其核心启动子区域位于-146～-8 bp区,该区域包含NFY、STAT1和cMYB等转录因子结合位点.前期研究结果提示,转录因子NFY可能参与NPCEDRG基因的转录调控.为明确NFY转录因子在NPCEDRG基因转录中的作用,本研究应用报告基因载体系统分析方法对NPCEDRG基因核心启动子区进行NFY结合位点(或CCAAT-box)缺失突变及其功能分析,分别构建NPCEDRG基因核心启动子区NFY结合位点缺失突变的Luc和EGFP报告基因表达载体.比较核心启动子和NFY结合位点缺失突变体报告基因载体的转录活性和效率.结果表明,在MCF7和CNE2细胞中,NFY结合位点缺失突变体的转录活性分别约为其核心启动子转录活性66.94%和75.72%,即NFY结合位点缺失致使该启动子转录效率在MCF7和CNE2细胞中分别降低了33.06%和24.28%;EGFP报告基因表达载体系统研究结果与Luc报告基因载体系统结果基本吻合.本研究再次证实,转录因子NFY通过结合NPCEDRG基因启动子的核心元件NFY结合位点参与NPCEDRG基因的转录调控,并提高其基因转录活性和效率.
[Abstract]:NPCEDRG gene is a nasopharyngeal carcinoma (nasopharyngeal carcinoma,NPC) related gene, NPCEDRG gene promoter is TATA-less promoter, its core promoter region is located in -146 bp region, which contains transcription factor binding sites such as NFY,STAT1 and cMYB. Previous studies suggest that transcription factor NFY may be involved in the transcriptional regulation of NPCEDRG gene. In order to clarify the role of NFY transcription factors in the transcription of NPCEDRG gene, the deletion mutation of NFY binding site (or CCAAT-box) and its function in the core promoter region of NPCEDRG gene were analyzed by the method of reporter gene vector system analysis. Luc and EGFP reporter gene expression vectors with deletion mutation of NFY binding site in core promoter of NPCEDRG gene were constructed. To compare the transcriptional activity and efficiency of core promoter and NFY binding site deletion mutants. The results showed that the transcriptional activities of NFY binding site deletion mutants in MCF7 and CNE2 cells were 66.94% and 75.72%, respectively. The loss of NFY binding site resulted in a reduction of transcription efficiency of the promoter by 33.06% and 24.28% in MCF7 and CNE2 cells, respectively. The results of EGFP reporter gene expression vector system were consistent with those of Luc reporter gene vector system. This study confirmed that transcription factor NFY participates in the transcriptional regulation of NPCEDRG gene by binding to the NFY binding site of the core component of the promoter of the NPCEDRG gene and enhances its transcriptional activity and efficiency.
【作者单位】：湖南师范大学医学院生物化学与分子生物学教研室;中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室;
【基金】：湖南省自然科学基金项目(No.13JJ3059) 湖南师范大学博士启动基金(No.130618)~~
【分类号】：R341

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