鳜SIRT3蛋白在传染性脾肾坏死病毒感染中的作用  

Role of the SIRT3 protein in spleen and kidney necrosis virus infections in Siniperca chuatsi

作　　者：李科瑾 付小哲[2] 林强[2] 刘礼辉[2] 牛银杰 梁红茹[2] 罗霞[2] 李宁求[2] LI Kejin;FU Xiaozhe;LIN Qiang;LIU Lihui;NIU Yinjie;LIANG Hongru;LUO Xia;LI Ningqiu(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Aural Affairs,Guangdong Provincial Key Laboratory of Aquatic Animal Immune Technology,Guangzhou 510380,China)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]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

出　　处：《中国水产科学》2021年第5期550-560,共11页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(31872589,U1701233);中国水产科学研究院基本科研业务费项目(2020XT0402);广东省现代农业产业技术创新团队专项基金项目(2019KJ140,2019KJ141)。

摘　　要：沉默交配型信息调节因子2同源蛋白3(silent mating type information regulation 2 homolog 3,SIRT3)是一种去乙酰化酶,可调节线粒体氧化代谢。为探讨鳜(Siniperca chuatsi)SIRT3在传染性脾肾坏死病毒(infectious spleen and kidney necrosis virus,ISKNV)感染中的作用,本研究克隆分析了鳜SIRT3基因,采用qRT-PCR和western blotting方法检测了病毒感染后SIRT3基因的表达,并探讨了SIRT3对细胞代谢酶基因的表达和病毒增殖的影响。结果显示,鳜SIRT3基因的开放阅读框全长1350 bp,编码449个氨基酸,在鳜各组织均表达,其中后肾中表达量最高,脾脏中表达量较低;ISKNV感染鳜脑细胞系和鱼体后,SIRT3表达量均显著升高;使用SIRT3抑制剂3-TYP(10μmol/L)和siRNA处理细胞后,谷氨酰胺酶(glutaminase,GLS)和谷氨酸脱氢酶(glutamatedehy drogenase,GDH)的表达量以及ISKNV产量均显著降低;使用SIRT3激动剂honokiol(7μmol/L)处理细胞后,代谢酶表达量和ISKNV产量均显著升高。结果表明,ISKNV感染可通过SIRT3诱导宿主细胞谷氨酰胺代谢重编程,进而促进自身增殖。Siniperca chuatsi is an economically important freshwater fish species cultivated in China.Infectious spleen and kidney necrosis virus(ISKNV)is the main causative agent of disease in S.chuatsi and causes huge economic losses in aquaculture.Viruses depend on cellular metabolisms for rapid replication and proliferation,and metabolomics analyses have revealed that the carbon flux of glutamine metabolism increases in ISKNV-infected cells.Previously,ISKNV has been reported to up-regulate the expression of glutaminase(GLS)and glutamate dehydrogenase(GDH)genes in host cells.These enzymes mediate glutamine metabolism and are necessary for the production of high-titer ISKNV particles.However,the mechanism of ISKNV-induced glutamine metabolism remains unclarified.The silent mating type regulation 2 homolog 3(SIRT3)is an NAD+-dependent deacetylase that regulates cellular metabolic conditions and participates in biological activities including cell cycle,mitochondrial ageing,and energy production.SIRT3 has been widely reported to respond to nutrient deficiencies and participate in cancer progression by regulating glutamine metabolism.The metabolic status of virus-infected cells is very similar to that of cancer cells,suggesting that SIRT3 may play a regulatory role during ISKNV infection.To clarify the potential mechanistic link between ISKNV and glutamine metabolism,we analyzed cloned sequences of S.chuatsi SIRT3 and detected expression of SIRT3 in different tissues of S.chuatsi.Then,the time-dependent expression patterns of the S.chuatsi SIRT3 in vitro and in vivo after ISKNV infection were assessed using real-time fluorescent quantitative PCR(qRT-PCR)and western blotting.Finally,the effects of SIRT3 on glutamine metabolism and the proliferation of ISKNV in host cells were analyzed by regulating the expression and the activity of SIRT3.Results showed that the S.chuatsi SIRT3 encoded 449 amino acids that showed high conservation.Expression of SIRT3 in 12 tissues of S.chuatsi was detected by qRT-PCR,with the maximum expressi

关 键 词：鳜 沉默交配型信息调节因子2同源蛋白3(SIRT3) 传染性脾肾坏死病毒(ISKNV) 谷氨酰胺代谢 

分 类 号：S941[农业科学—水产养殖]