N-甲基-N’-硝基-N-亚硝基胍诱导共表达的蛋白基因启动子区的转录因子结合部位分析  

Analysis of the transcription factor binding sites in the promoter region of genes encoding the co-expressive proteins induced by N-methyl-N'-nitro-N-nitrosoguanidine

作　　者：刘天婵[1] 余应年[1] 

机构地区：[1]浙江大学医学院病理生理学教研室,浙江杭州310031

出　　处：《中国病理生理杂志》2004年第11期1953-1960,共8页Chinese Journal of Pathophysiology

基　　金：国家重点基础研究发展规划项目 (No .2 0 0 2CB5 12 90 1);国家自然科学基金资助项目 (No.30 170 810 )

摘　　要：目的 :分析甲基硝基亚硝基胍 (MNNG)诱导的共表达蛋白的编码基因启动子区的转录因子结合部位。方法 :用进化足迹法 ,结合转录因子数据库的搜索 ,预测共表达蛋白编码基因启动子区共有的转录因子结合部位。凝胶阻滞试验验证对于预测出的转录因子结合部位 ,在MNNG处理细胞中是否确实有相关转录因子的反应。结果 :预测出 11个共同存在于这些蛋白编码基因启动子区的转录因子结合部位 ,其中除已知转录因子激活蛋白 1(activatorprotein 1,AP1)在MNNG处理后被激活外 ,用凝胶阻滞试验又发现在MNNG处理细胞的核提取液中有 2个转录因子—核因子Y(nuclearfactor,NFY)和GATA结合因子 (GATAbindingfactor,GATA)的活性升高。结论 :进化足迹法可以有效地降低转录因子结合部位预测结果中的假阳性率。NFY和GATA转录因子结合部位可能参与对MNNG诱导的共表达蛋白的共调控。AIM: To find out common transcription factor binding sites in the promoter regions of the encoding genes of the co-expressive proteins induced by N-methyl-N'-nitro-N- nitrosoguanidine (MNNG). METHODS: Using phylogenetic footprinting and TRANSFAC position weight matrix (PWM) searching program to predict the common transcription factor binding sites among the promoter regions of the genes encoding the co-expressive proteins. The predictive results were validated with electrophoresis mobility shift assay (EMSA). RESULTS: Eleven common transcription factor binding sites were predicted in the promoters of the co-expressive proteins, among them, besides the activator protein 1(AP1) which was previously identified to be activated in MNNG pretreated cells in this laboratory, the nuclear factor Y (NFY) and GATA binding factor (GATA) consensus oligonucleotides binding activity were found being increased in the nuclear extract of cells pre-treated with MNNG as demonstrated by EMSA. CONCLUSION: Phylogenetic footprinting can effectively decrease the false positive rate in predicting transcription factor binding sites. It is possible that NFY and GATA transcription factor binding sites are involved in the co-regulation of the MNNG induced co- expressive proteins. [

关 键 词：进化足迹法 转录因子结合部位 凝胶阻滞试验 甲基硝基亚硝基胍 

分 类 号：R318[医药卫生—生物医学工程] Q75[医药卫生—基础医学]