端粒相关蛋白PinX1和磷酸化激酶Aurora A的相互作用  

Interaction study between telomere-related protein PinX1 and phosphorylated kinase Aurora A

作　　者：李梦亚 王冲[1] 张好好[2] 王树娟[1] 刘延方[1] 姜中兴[1] LI Mengya;WANG Chong;ZHANG Haohao;WANG Shujuan;LIU Yanfang;JIANG Zhongxing(Department of Hematology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052;Department of Endocrinology, the First Affiliated Hospital,Zhengzhou University, Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院血液科,郑州450052 [2]郑州大学第一附属医院内分泌及代谢病科,郑州450052

出　　处：《郑州大学学报（医学版）》2021年第1期38-42,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(U1804191,U1904137,81800137);河南省高等学校重点科研项目(20A320061);河南省重点研发与推广专项(182102310563)。

摘　　要：目的:鉴定端粒相关蛋白PinX1与有丝分裂期极光激酶(Aurora A)的相互作用及其功能,进一步研究PinX1调控参与细胞有丝分裂期的具体机制。方法:以pGBKT7-PinX1为诱饵对人HeLa细胞cDNA文库进行筛选,对筛选到的结果进行测序分析;构建Aurora A原核及真核表达质粒;采用免疫共沉淀及蛋白质体外沉降实验进行相互结合研究;采用体内磷酸化实验验证Aurora A是否能磷酸化修饰PinX1;免疫荧光染色实验检测Aurora A与PinX1是否存在共定位。结果:酵母双杂交结果发现了3个新的PinX1相互作用蛋白;免疫共沉淀及体外沉降实验证实PinX1与Aurora A存在相互结合;免疫荧光染色实验证实PinX1与Aurora A在细胞内存在共定位;磷酸化实验证实Aurora A能够磷酸化修饰PinX1。结论:Aurora A是一个新的PinX1结合蛋白;Aurora A能够磷酸化修饰PinX1;PinX1可能通过Aurora A磷酸化修饰参与细胞有丝分裂期调控。Aim:To identify the interaction and function of telomere-related protein PinX1 and mitotic phosphorylated kinase Aurora A,and to further study the specific mechanism of PinX1 involved in cell mitotic regulation.Methods:Screening cDNA library of human HeLa cells with pGBKT7-PinX1 as a bait and sequencing analysis of the results.Constructing prokaryotic and eukaryotic expression plasmids with Aurora A.To investigate the interaction of PinX1 and Aurora A using co-immunoprecipitation and protein pull-down assay in vitro.To verify whether Aurora A could phosphorylate PinX1 with in vivo phosphorylation assay.Immunofluorescence assay was used to detect the colocalization of Aurora A and PinX1.Results:Three new PinX1 interacting proteins were discovered in yeast two-hybrid assay.Co-immunoprecipitation and in vitro protein pull-down assay confirmed that Aurora A and PinX1 were bound to each other.Immunofluorescence assay affirmed the colocalization of Aurora A and PinX1 in cells.Phosphorylation assay verified PinX1 could be phosphorylated by Aurora A.Conclusion:Aurora A is a new PinX1 binding protein and it can phosphorylate PinX1.PinX1 may be involved in the regulation of cell mitosis through Aurora A phosphorylation.

关 键 词：PinX1 Aurora A 有丝分裂 端粒 

分 类 号：R730.231[医药卫生—肿瘤]