摘要
为解析塞内卡病毒(SVA)VP3蛋白对宿主细胞蛋白翻译系统的调控机制,首先利用激光共聚焦免疫荧光和蛋白免疫印迹试验,证实外源表达的SVA VP3蛋白能够诱导宿主细胞蛋白翻译阻滞;进一步利用荧光素酶报告基因检测、荧光定量PCR和蛋白免疫印迹等试验,发现VP3介导的翻译阻滞不依赖宿主细胞内蛋白降解途径和转录调控,而是通过诱导细胞内eIF2α发生磷酸化,抑制m TOR-S6K信号通路活化、eIF4E磷酸化和eIF4F复合体形成,导致宿主细胞蛋白合成效率严重受阻。结合互作蛋白组学数据显示,VP3蛋白在病毒劫持宿主蛋白翻译系统促进自身增殖的过程中发挥重要作用。综上所述,揭示了SVAVP3蛋白诱导宿主细胞翻译阻滞的潜在机制,为深入认识SVA翻译调控与感染机制提供了新视角。
To elucidate the regulatory mechanism of Senecavirus A(SVA)VP3 protein on the host cellular translation system,this study initially employed laser confocal immunofluorescence and Western-blot analysis to confirm that exogenous expression of SVA VP3 induced host cellular translation arrest.Further investigations utilizing luciferase reporter assays,quantitative real-time PCR(qRT-PCR),and Western-blot revealed that SVA VP3-mediated host translation arrest was independent of intracellular protein degradation pathways and transcriptional regulation.Instead,SVA VP3 not only triggers phosphorylation of eIF2αbut also suppresses mTOR-S6K signaling pathway activation,eIF4E phosphorylation,and eIF4F complex assembly,thereby severely impairing host cellular protein synthesis.Furthermore,proteomics analysis indicated that SVA VP3 played a pivotal role in viral hijacking of host translational machinery to facilitate viral proliferation.Collectively,this study elucidated the underlying mechanisms by which SVA VP3 protein induced host translational arrest,providing novel insights into understanding SVA translational regulation and pathogenesis of infection.
作者
邓小双
李鑫威
万浩成
周梦菡
胡曼
张雨杭
万博
韩世充
DENG Xiaoshuang;LI Xinwei;WAN Haocheng;ZHOU Menghan;HU Man;ZHANG Yuhang;WAN Bo;HAN Shichong(International Joint Research Center of National Animal Immunology,College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)
出处
《中国兽医科学》
北大核心
2025年第10期1310-1318,共9页
Chinese Veterinary Science
基金
国家自然科学基金面上项目(32373005)
河南省优秀青年科学基金项目(252300421156)。
