摘要
为分析富含脯氨酸核受体辅调节蛋白1(PNRC1)选择性剪接,及比较PNRC1剪接变异体在辅激活核受体介导基因转录功能上的差异,在生物信息学方法分析PNRC1剪接变异体的基础上,设计一定的特异性引物,采用RT-PCR结合克隆测序的方法对这些剪接变异体进行验证.利用酵母双杂交和荧光素酶报告系统实验,分析它们与核受体的相互作用及比较它们在辅激活核受体介导基因转录功能上的差异.结果显示,生物信息学预测的几个剪接变异体真实存在于人的组织和细胞系中,这些剪接变异体在与雌激素受体α(ERα)、类固醇衍生因子1(SF1)等核受体的相互作用的强度及辅激活核受体介导基因转录功能上存在较大的差异.研究提示,PNRC1这些剪接变异体在体内可能发挥不同的功能.
To identify and characterize splicing variants of proline-rich nuclear receptor coregulator protein 1 （ PNRC1 ）, we firstly tested whether PNRC1 splice variants predicted by informatics were naturally occurring in human tissues or cell lines, secondly analyzed their interactions with nuclear receptors such as estrogen receptor alpha （ERa） and steroidogenic factor 1 （SF1） by yeast two-hybrid assay, and compared their abilities to potentiate nuclear receptors ERα or SF1 mediated transcription using luciferase reporter gene system. The results showed that most of the PNRC1 splice variants predicted by bioinformatics were naturally occurring in human tissues or cell lines, and they had great differences in interacting with ERα or SF1 and enhancing these nuclear receptors mediated transcriptions. The results suggest that PNRC1 splicing variants may exert different functions in vivo.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2007年第12期1019-1024,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然基金资助项目(No.30671056)~~
