Purα蛋白质不同结构域对APP基因表达的调控作用  

Effects of Purα or Its Domains on the Regulation of APP Gene Expression

作　　者：李永玲[1] 贾中发 柴娟[2] 李平[1] 袁程敏 孙涛[2] 崔建奇[2,1] 

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]宁夏医科大学颅脑疾病重点实验室省部共建国家重点实验室培育基地,银川750004

出　　处：《中国细胞生物学学报》2015年第9期1278-1287,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81260197);国家重点基础研究发展计划(973计划)(批准号:2012CB722408;2014CB560711)资助的课题~~

摘　　要：该研究将构建的含淀粉样前体蛋白(amyloid precursor protein,APP)基因启动子的萤火虫荧光素酶报告质粒与Purα全长基因或含不同结构域的Purα缺失突变体共转染到U87MG细胞中,进行萤火虫荧光素酶活性测定,以确定Purα对APP基因表达的调控作用。同时,将Purα全长基因及含有不同结构域的Purα缺失突变体的真核表达载体分别转染至U87MG细胞,使目的蛋白过表达,通过实时定量PCR(Real-time PCR)和蛋白免疫印迹(Western blot)研究Purα不同的结构域对APP基因在转录、翻译水平的调控作用。结果显示,Purα全长蛋白及其不同结构域对APP基因表达均有不同程度的抑制作用,但至少保持N-端或C-端的结构域可能是维持Purα蛋白功能的必要条件。The eukaryotic expression vector pCDNA3.0-Pure and the deletions which contain the different DNA binding domains were constructed. Then co-transfected into U87MG cells with Luciferase reporter construct with which the APP promoter （-170/＋147） was inserted into the upstream of Luciferase gene. The Luciferase assays were performed to analyze the transactivation effects of Pure on APP gene expression. The Real-time PCR and Western blot were used to evaluate the effects of Pura and the different domains on APP gene expression both in transcriptional and translational levels. The results of Luciferase assay demonstrated that the full length of Pura and the different deletions could down-regulate APP promoter activities in various extent; at the meantime, the results of Real-time PCR and Western blott confirmed that Pura could suppress APP gene expression both in transcriptional and translational levels, but at least to keep the intact structure in N-terminal or C-terminal of the Pure may be essential for maintenance of the functions of Pura.

关 键 词：Purα 蛋白质结构 结构域 APP 基因表达 

分 类 号：Q78[生物学—分子生物学]