人CtBP2与CCNH相互作用的初步研究

Preliminary Research into the Interaction between Human CtBP2 and CCNH

CtBP2(E1AC-terminal binding protein 2)作为辅阻遏物与多种转录因子联系而参与到很多生物过程中,如细胞分化、凋亡、发育和肿瘤发生等,然而其中许多作用机制尚不明了.为了对CtBP2进行深入研究,利用高通量酵母双杂交技术,以人CtBP2为诱饵,与含有1000个人肝基因克隆的酵母双杂交文库进行接合筛选获得了一个与它相互作用的猎物蛋白CCNH(Cyclin H).通过GST-pull down、免疫共沉淀和亚细胞共定位等实验进一步证明了这两个蛋白在体外和体内的相互作用.

CtBP2（E1A C-temainal binding protein 2） has linked to multiple biological precesses through their association with numerous transcription factors. It plays a crucial role in differentiation, apoptosis, development and oncogenesis. However, how it represses the transcription is not clear. For further study, the high through-put yeast two hybrid system was used to do large scale mating with the library of 1000 yeast clones containing genes of human liver proteins by using Human CtBP2 as a bait. CCNH which interacts with CtBP2 was identified. Pull-down and coimmunoprecipitation were used to confirm this interaction. The co-loclization assay showed that CtBP2 co-lccalizes with CCNH in the nucleus.