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Effects of Purα or Its Domains on the Regulation of APP Gene Expression
Li Yongling2#, Jia Zhongfa2#, Chai Juan1, Li Ping2, Yuan Chengmin1, Sun Tao1, Cui Jianqi1,2*
1Ningxia Key Laboratory of Cerebrocranial Diseases, the Incubation Base of National Key Laboratory, Yinchuan 750004, China;
2School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China
2School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China
Abstract: The eukaryotic expression vector pCDNA3.0-Purα 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 Purα on APP gene expression. The Real-time PCR and Western blot were used to evaluate the effects of Purα 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 Purα 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 Purα 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 Purα may be essential for maintenance of the functions of Purα.
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