hCDC14A与BRAP2相互作用的初步研究

目的： 筛选hCDC14A(human cell division cycle gene 14A)的相互作用蛋白，探索功能联系。方法： 酵母双杂交筛选hCDC14A的可能相互作用蛋白，Pulldown实验和免疫共沉淀实验验证蛋白质间相互作用，免疫荧光显微镜观察目的基因在细胞中的表达和定位，体内泛素化实验提示可能的相互作用机制。结果： 酵母双杂交筛查到BRAP2(BRCA1 associated protein 2)为hCDC14A的可能相互作用蛋白，两者有共同定位，BRAP2可以增强hCDC14A的泛素化修饰。结论： BRAP2可以与hCDC14A相互作用，可能是hCDC14A的泛素连接酶。

Primary analysis on interaction between hCDC14A and BRAP2

AIM: To search for new candidate partners of human cell division cycle gene 14A (hCDC 14A) and explore their functional relationships.METHODS: Yeast two-hybrid screening was employed to find hCDC 14A new partners. Physical interaction between two proteins was verified using Pulldown and co-immunoprecipitation assays. Subcellular localizations were revealed by immunofluorescence. In vivo ubiquitination test implied their potential functional relationship.RESULTS: BRAP2 (BRCA1 associated protein 2) was found to be a new candidate partner of hCDC 14A. hCDC 14A was modified by ubiquitination, and BRAP2 increased this modification in vivo. As expected, hCDC 14A and BRAP2 co-localized on mitotic spindles in HeLa cells.CONCLUSION: BRAP2 may be an ubiquitin E3 ligase of hCDC 14A.