鳜弹状病毒调控谷氨酰胺还原性代谢途径促进自身增殖  

Reductive glutamine metabolism promotes the efficient replication of Siniperca chuatsi rhabdovirus in Chinese perch brain cells

作　　者：郭茜茜 付小哲[1] 梁红茹[1] 林强[1] 刘礼辉[1] 牛银杰 李宁求[1] GUO Xixi;FU Xiaozhe;LIANG Hongru;LIN Qiang;LIU Lihui;NIU Yinjie;LI Ningqiu(Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Guangdong Provincial Key Laboratory of Aquatic Animal Immune Technology,Guangzhou 510380,China;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]中国水产科学研究院珠江水产研究所,农业农村部渔用药物创制重点实验室,广东省水产动物免疫技术重点实验室,广东广州510380 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《中国水产科学》2019年第5期993-1003,共11页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(31872589);广东省渔港建设和渔业产业发展专项(粤财农[2017]17号);广东省促进经济发展专项资金(海洋经济发展用途)(GDME-20181007);广州市珠江科技新星项目(201710010087)

摘　　要：为探究鳜弹状病毒(Siniperca chuatsi rhabdovirus,SCRV)复制增殖与谷氨酰胺还原性代谢(reductive glutamine metabolism,RGM)途径的相互作用关系,以鳜脑细胞系(Chinese perch brain cells,CPB)为增殖体系,采用实时荧光定量PCR技术检测病毒拷贝数和RGM途径关键酶mRNA表达水平变化,并通过蛋白免疫印迹方法检测RGM途径关键酶蛋白表达水平变化。结果发现,当培养基缺失谷氨酰胺时SCRV拷贝数降低了99.5%,表明谷氨酰胺是SCRV增殖所必需;而SCRV感染上调CPB细胞RGM途径关键酶的表达,其中异柠檬酸脱氢酶2(isocitrate dehydrogenase 2,IDH2)上调倍数最大,表明SCRV感染上调了细胞RGM途径中关键酶基因的表达;同时,抑制细胞RGM途径关键酶的表达导致SCRV病毒产量显著下降,表明RGM途径有利于SCRV复制增殖;进一步敲降细胞IDH2基因表达可显著抑制SCRV N蛋白表达,而过表达细胞IDH2基因可显著促进SCRV N蛋白表达,表明IDH2在病毒增殖中具有重要作用。综上所述,SCRV感染可调控CPB细胞RGM通路以满足其自身增殖,其结果可为阐明鳜弹状病毒病致病机制和抗病毒治疗策略提供新的思路。Viruses rely on host cellular metabolism for energy and macromolecule synthesis during replication.Therefore,there is a close relationship between viral replication and cellular metabolism.Siniperca chuatsi rhabdovirus(SCRV)causes significant economic losses in the Chinese perch(Siniperca chuatsi)industry.Thus,it is important to address the interaction between SCRV replication and host cells to prevent and control SCRV.However,little is known about the relationship between SCRV replication and cellular metabolism.In addition,glutamine is an abundant amino acid necessary for energy generation and macromolecule synthesis in cells.Glutamine is also essential for the infection and replication of some viruses.Glutaminolysis and reductive glutamine metabolism(RGM)are the main pathways of glutamine metabolism.Particularly,the RGM pathway plays an important role in tumor cells.However,the metabolic pathway of glutamine involved in viral replication remains largely uncharacterized.To clarify the interaction between SCRV replication and RGM,we analyzed the expression of key enzymes in the RGM pathway in Chinese perch brain(CPB)cells following infection with SCRV,using quantitative real-time PCR(qRT-PCR)and western blotting.Our results showed that SCRV replication was decreased in CPB cells cultured in a glutamine-depleted medium,indicating that glutamine is required for efficient SCRV replication.In addition,SCRV infection promoted the expression of key enzymes involved in the RGM pathway in CPB cells,particularly the expression of isocitrate dehydrogenase 2(IDH2),indicating that RGM in CPB cells is altered during SCRV infection.SCRV proliferation was also inhibited in CPB cells cultured in the presence of bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl)ethyl sulfide(a glutaminase inhibitor),(–)-epigallocatechin monogallate(a glutamate dehydrogenase inhibitor),5-(tetradecyloxy)-2-furoic acid(an acetyl-CoA carboxylase inhibitor),or 4-methylene-2-octyl-5-oxotetrahydrofuran-3-carboxylic acid(a fatty acid synthase inhibitor),

关 键 词：鳜脑细胞系 鳜弹状病毒 谷氨酰胺还原性代谢途径 

分 类 号：S941[农业科学—水产养殖]