人E1A激活基因阻遏子基因启动子生物信息学分析及转录调控功能初探  

Bioinformatic analysis of the cellular repressor of E1A-stimulated genes proximal promoter and primary identification of transcriptional regulation function

作　　者：韩雅玲[1] 赵昕[1] 闫承慧[1] 康建[1] 张效林[1] 邓捷[1] 徐红梅[1] 刘海伟[1] 

机构地区：[1]沈阳军区总医院全军心血管病研究所心内科,辽宁沈阳110016

出　　处：《中国实用内科杂志》2007年第12期935-937,共3页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金(30570664);辽宁省自然科学基金(2050426)

摘　　要：目的对人类腺病毒5型早期区域1A(adenovirus type 5 early region 1A,E1A)激活基因阻遏子基因(cellular repressor of E1A-stimulated genes,hCREG)启动子进行生物信息学分析、预测hCREG启动子位置和转录因子结合位点。方法对2005年1月至7月沈阳军区总医院全军心血管病研究所进行的从Genbank中获得hCREG的全长编码基因,利用First EF程序分析软件分析并获得hCREG的启动子序列,利用Motif软件对启动子序列中的转录因子结合位点进行预测。免疫荧光染色和Western blot分别观察相关转录因子的蛋白表达与细胞内hCREG蛋白及细胞表型转化之间的关系。结果用First EF软件测定长度为945bp的hCREG启动子序列,hCREG的启动子区可能定位于转录起始点上游-109^-359bp。用Motif软件预测启动子区域共含有80多个转录调控因子结合位点,其中肿瘤抑制基因wtp53结合位点为-456bp。进一步应用蛋白质印迹(western blot)和免疫荧光染色分析wtp53与细胞表型转化的关系。在0.5%胎牛血清(fetalbovine serum,FBS)培养的HITASY细胞wtp53表达较10%FBS培养HITASY细胞明显增加。同时,hCREG蛋白和平滑肌细胞分化标志蛋白SMα-ac-tin亦表达增多。结论获得了hCREG启动子的转录调控信息,推测出wtp53是hCREG基因转录的重要调控因子,它可能结合于hCREG启动子区,参与hCREG蛋白对VSMCs表型转化的调控。Objective To analyze the human cellular repressor of E1A-stimulated genes （hCREG） using bioinformatics tools and predict the promoter position and transciption factor binding sites. Methods Complete coding hCREG sequences were obtained from Genbank. hCREG was analyzed with First EF software in order to obtain the promoter sequence of hCREG. Moreover, transcription factor binding sites of hCREG was convinced by Motif software. Subsequently, the transcription factor of hCREG by bioinformatics tools was related between hCREG and smooth muscle cells differentiation observed by both Western blot and immunoflourescence. Results The promoter of hCREG was located in - 109 ~ -359 bp of up-stream of transcriptional site,which was predicted with 945bp length. Transciption factor binding sites of hCREG was predicted by Motif software which was found with 80 transciption factors. The expression of hCREG and smooth muscle α-actin（ SM α-actin）increased in HITASY after 72 hours with serum deprivation detected by both Western blot and immunoflourescence. Meanwhile, the results showed that the expression of wtp53 increased significantly. These resuits suggested that transcription factor wtp53 might regulate the expression of hCREG and promoted dedifferentiated phenotvpe of VSMCs. Conclusion The transcriptional information of the proximal promoter of hCREG obtained. As up-stream regulational factor of hCREG, wtp53 can up-regulate the expression of hCREG and may play a vital role in the process of VSMCs differentiation and phenotype modulation.

关 键 词：启动子 生物信息学 WTP53 阻遏蛋白 E1A 表型 

分 类 号：R5[医药卫生—内科学]