SYVN1泛素化降解XRCC5对晶状体上皮细胞DNA氧化损伤的影响  

Effects of SYVN1-mediated ubiquitination and degradation of XRCC5 on DNA oxidative damage in lens epithelial cells

作　　者：王从玉 李鹏飞 王思文 鲍思洁 石海红[1] 管怀进[1] WANG Congyu;LI Pengfei;WANG Siwen;BAO Sijie;SHI Haihong;GUAN Huaijin(Eye Institute,Affiliated Hospital of Nantong University,Medical School of Nantong University,Nantong 226001,Jiangsu Province,China)

机构地区：[1]南通大学附属医院眼科,南通大学医学院,江苏省南通市226001

出　　处：《眼科新进展》2023年第10期761-765,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金面上项目(编号:82171038,81974129)。

摘　　要：目的探索E3泛素连接酶SYVN1在人晶状体上皮细胞系SRA01/04中对DNA氧化损伤的影响。方法以中波紫外线(UVB)处理的SRA01/04细胞为研究对象。将培养的SRA01/04细胞分为对照组、OV-SYVN1组、UVB组、UVB+HA组和UVB+OV-SYVN1组,应用Western blot实验检测SYVN1和XRCC5蛋白的相对表达量,应用Western blot实验检测DNA损伤标志物γH2A蛋白的相对表达量,应用免疫荧光染色检测DNA损伤标志物γH2A的荧光强度变化。应用免疫沉淀实验验证XRCC5的泛素化修饰及其与SYVN1的相互作用。结果Western blot实验结果表明,与对照组比较,OV-SYVN1组中SYVN1蛋白相对表达量明显上调(P<0.01);与对照组比较,UVB组中γH2A蛋白相对表达量明显升高(P<0.0001);与UVB+HA组比较,UVB+OV-SYVN1组中γH2A蛋白相对表达量明显升高(P<0.01)。免疫荧光染色结果表明,与UVB+HA组比较,UVB+OV-SYVN1组中γH2A染色明显加强。UbiBrowser软件预测发现,SYVN1可能是XRCC5潜在的E3泛素连接酶;免疫沉淀实验结果显示,SYVN1和XRCC5存在相互作用,促进XRCC5蛋白发生泛素化修饰。Western blot实验结果显示,与对照组比较,OV-SYVN1组中XRCC5蛋白相对表达量明显下降(P<0.001)。结论SYVN1可能介导DNA氧化损伤修复蛋白XRCC5产生泛素化修饰,并通过降解XRCC5蛋白表达参与DNA氧化损伤的修复过程,调控年龄相关性白内障的疾病进程。Objective To explore the effects of E3 ubiquitin ligase SYVN1 on DNA oxidative damage in human lens epithelial cell line SRA01/04.Methods The SRA01/04 cells treated with medium-wave ultraviolet(UVB)were divided into the control group,OV-SYVN1 group,UVB group,UVB+HA group and UVB+OV-SYVN1 group.The relative expression levels of SYVN1,XRCC5,and DNA damage markerγH2A proteins were measured by Western blot.The changes in the fluorescence intensity of theγH2A were detected by immunofluorescence staining.The ubiquitination of XRCC5 and its interactions with SYVN1 were verified by immunoprecipitation.Results Western blot experiments showed that compared with the control group,the expression of SYVN1 protein in the OV-SYVN1 group was significantly up-regulated(P<0.01);compared with the control group,the relative expression ofγH2A protein in the UVB group significantly increased(P<0.0001);compared with the UVB+HA group,the relative expression ofγH2A protein in the UVB+OV-SYVN1 group significantly increased(P<0.01).Immunofluorescence staining results showed that compared with the UVB+HA group,γH2A staining was significantly enhanced in the UVB+OV-SYVN1 group.UbiBrowser software predicted that SYVN1 might be a potential E3 ubiquitin ligase of XRCC5.The immunoprecipitation experiments proved that there was indeed an interaction between SYVN1 and XRCC5,where SYVN1 promoted the ubiquitination of XRCC5 protein.Western blot experiments showed that compared with the control group,the relative expression of XRCC5 protein in the OV-SYVN1 group significantly decreased(P<0.001).Conclusion SYVN1 may mediate the ubiquitination of DNA oxidative damage repair protein XRCC5,participate in the repair process of DNA oxidative damage by degrading the expression of XRCC5 protein,and regulate the age-related cataract process.

关 键 词：年龄相关性白内障 泛素化修饰 DNA损伤修复 

分 类 号：R776[医药卫生—眼科]