XRCC6调节RYBP表达的分子机制与功能  

Molecular Mechanism and Function of XRCC6 Regulates RYBP Expression

作　　者：刘薇佳 李唐嫒 陈修远 夏婉娉 陈等 LIU Weijia;LI Tangai;CHEN Xiuyuan(Institute of Basic Medical Sciences Chinese Academy of Medical Sciences,School of Basic Medicine Peking Union Medical College,Department of Biochemistry and Molecular Biology,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所、北京协和医学院基础学院生物化学与分子生物学系,100005

出　　处：《医学研究杂志》2024年第12期31-36,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81272229);中国医学科学院创新工程项目(2021-I2M-1-016,2017-I2M-3-004)。

摘　　要：目的研究X-射线修复交叉互补蛋白6(X-ray repair cross complementing 6,XRCC6)结合和下调RING1和YY1结合蛋白(ring1 and YY1 binding protein,RYBP)表达的分子机制与功能。方法采用蛋白质免疫共沉淀和免疫荧光方法分析XRCC6与RYBP之间的相互作用;采用Western blot法检测XRCC6对RYBP表达水平的影响;采用Western blot法和蛋白质免疫共沉淀法检验XRCC6对RYBP蛋白稳定性的调节作用;采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)检测XRCC6是否通过RYBP调节基因表达。结果内源性RYBP与XRCC6之间存在相互作用;XRCC6负性调节RYBP蛋白水平;过表达XRCC6促进RYBP的多聚泛素化和蛋白酶体降解,显著缩短其半衰期;XRCC6负调节H2AK119Ub1,通过调节RYBP抑制下游基因的表达。结论XRCC6结合并通过蛋白酶体途径下调RYBP表达,进而调节RYBP下游基因的表达。Objective To investigate the molecular mechanism and function of X-ray repair cross complementing 6(XRCC6)regulate Ring1 and YY1 binding protein(RYBP)expression.Methods The interaction between XRCC6 and RYBP was analyzed by protein co-immunoprecipitation and immunofluorescence assays.The effect of XRCC6 on RYBP protein level was determined by Western blotting.The influence of XRCC6 on the stability of RYBP was detected by Western blotting and protein co-immunoprecipitation assays.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to determine whether XRCC6 regulates gene expression through RYBP.Results There was an interaction between endogenous RYBP and XRCC6,and XRCC6 negatively regulated the level of RYBP protein.Overexpression of XRCC6 promoted polyubiquitination and proteasome degradation of RYBP,and significantly shortened its half-life.Furthermore,XRCC6 negatively regulated H2AK119Ub1 level and inhibited downstream gene expressions through modulating RYBP.Conclusion XRCC6 interacts with and down-regulates RYBP expression through proteasome pathway,thereby regulating the expression of RYBP downstream genes.

关 键 词：XRCC6 RYBP 蛋白质相互作用 蛋白酶体途径 基因表达调节 

分 类 号：R393[医药卫生—基础医学]