TMEM59的羧基端片段经与ATG5-ATG16L1-LC3B蛋白相互作用促进自噬  

The carboxy-terminal fragment of TMEM59 promotes autophagy via interaction with ATG5-ATG16L1-LC3B proteins

作　　者：刘肇绩[1] 郑晓龄 黄榕 董佳萌 吴紫微 彭锋[1] LIU Zhaoji;ZHENG Xiaoling;HUANG Rong;DONG Jiameng;WU Ziwei;PENG Feng(Department of Neurology,Zhongshan Hospital Affiliated to Xiamen University,Fujian,Xiamen 361004,China;The Third Clinical Medical College,Fujian Medical University,Fujian,Fuzhou 350000,China;School of Medicine,Xiamen University,Fujian,Xiamen 361004,China)

机构地区：[1]厦门大学附属中山医院神经内科,福建厦门361004 [2]福建医科大学第三临床医学院,福建福州350000 [3]厦门大学医学院,福建厦门361004

出　　处：《中国医药科学》2024年第19期8-11,共4页China Medicine And Pharmacy

基　　金：福建省自然科学基金项目(2021J011339);福建省厦门市医疗卫生指导性项目(35022202242D1042)。

摘　　要：目的研究跨膜蛋白59(TMEM59)诱导自噬、TMEM59诱导自噬的部位及探讨TMEM59诱导自噬的相关通路。方法通过细胞转染、免疫共沉淀、免疫荧光等技术明确TMEM59能否诱导自噬及诱导自噬的部位,自噬流抑制剂处理明确TMEM59诱导自噬的相关通路。结果研究证实在Hela细胞中过表达TMEM59和TMEM59的羧基端片段(CTF)增加自噬标记蛋白微管相关蛋白1轻链3(LC3B)-Ⅱ的水平以及促进自噬流的发生,差异有统计学意义(P<0.05),但过表达TMEM59的氨基端片段(NTF)差异无统计学意义(P>0.05)。过表达TMEM59不会影响自噬激活剂雷帕霉素诱导的LC3B-Ⅱ水平增加,差异无统计学意义(P>0.05),促进因抑制自噬溶酶体融合所导致的LC3B-Ⅱ水平增加,差异有统计学意义(P<0.05)。证实TMEM59、TMEM59-CTF与自噬相关基因蛋白(ATG)5、ATG16L1和LC3B相互作用。结论TMEM59-CTF诱导自噬发生在自噬起始之后和自噬溶酶体形成之前的阶段,与ATG5-ATG16L1-LC3B蛋白相互作用促进自噬。Objective To investigate the induction of autophagy by transmembrane protein 59(TMEM59)and the site of autophagy induced by TMEM59,and to explore the related pathways of autophagy induced by TMEM59.Methods The ability of TMEM59 to induce autophagy and the site of autophagy induction were clarified using techniques such as cell transfection,Co-immunoprecipitation,immunofluorescence,etc.The related pathways of autophagy induced by TMEM59 were clarified through treatment with inhibitors of autophagy flux.Results Studies confirmed that overexpression of TMEM59 and the carboxy-terminal fragment(CTF)of TMEM59 in cells increased the level of autophagy marker protein microtubule-associated protein 1 light chain 3(LC3B)-Ⅱand promoted the occurrence of autophagy flux,with statistically significant differences(P<0.05).However,the difference in the overexpression of amino-terminal fragment(NTF)of TMEM59 was not statistically significant(P>0.05).Overexpression of TMEM59 did not affect the increase in the level of LC3B-Ⅱinduced by the autophagy activator rapamycin,with no statistically significant difference(P>0.05).But it promoted the increase in the level of LC3B-Ⅱdue to the inhibition of autolysosome fusion,with a statistically significant difference(P<0.05).TMEM59 and TMEM59-CTF were confirmed to interact with the autophagy-associated gene protein(ATG)5,ATG16L1 and LC3B.Conclusion TMEM59-CTF can induce autophagy at a stage after the initiation of autophagy and before the formation of autolysosome,and interact with ATG5-ATG16L1-LC3B proteins to promote autophagy.

关 键 词：跨膜蛋白59 自噬 自噬标记蛋白微管相关蛋白1轻链3 信号通路 

分 类 号：R741[医药卫生—神经病学与精神病学]