【摘要】 目的:筛选hCDC14A(human cell division cycle gene14A)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 protein2)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。更多还原

【Abstract】 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.更多还原