Glutamine and glutaminolysis are required for efficient replication of infectious spleen and kidney necrosis virus in Chinese perch brain cells

// Xiaozhe Fu 1, 2, 4, * , Xianqin Hu 2, 3, * , Ningqiu Li 1, 2 , Feifei Zheng 2 , Xingxing Dong 2 , Jing Duan 2 , Qiang Lin 1, 2 , Jiagang Tu 2 , Lijuan Zhao 2 , Zhibin Huang 1 , Jianguo Su 2, 4 , Li Lin 2, 5 1 Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Fishery Drug Development, Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology, Guangdong Provinces, Guangzhou, Guangdong, 510380, China 2 Department of Aquatic Animal Medicine, Research Center of Marine Biology, College of Fisheries, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province Huazhong Agricultural University, Wuhan, Hubei, 430070, China 3 School of Animal Sciences and Nutritional Engineering, Wuhan Polytechnic University, Wuhan, Hubei, 430023, China 4 College of Animal Science and Technology, Northwest A and F University, Shanxi Key Laboratory of Molecular Biology for Aquaculture, Yangling, 712100, China 5 State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, Haikou 570228, China * These authors contributed equally to the work Correspondence to: Li Lin, email: linli@mail.hzau.edu.cn Ningqiu Li, email: liningq@126.com Keywords: Siniperca chuatsi, ISKNV, glutamine, glutaminolysis, TCA cycle Received: April 24, 2016      Accepted: November 21, 2016      Published: November 29, 2016 ABSTRACT Viruses rely on host cellular metabolism for energy and macromolecule synthesis during their replication. Infectious spleen and kidney necrosis virus (ISKNV) causes significant economic losses in the Chinese perch ( Siniperca chuatsi ) industry worldwide. However, little is known about the relationship between ISKNV replication and cellular metabolism. Using transcriptomic analysis, we observed that glutamine metabolism in Chinese perch brain (CPB) cells is altered during ISKNV infection. Moreover, ISKNV replication was decreased in CPB cells cultured in the glutamine-depleted medium. ISKNV replication was also inhibited in CPB cells cultured in the presence of bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl) ethyl sulfide (an inhibitor of glutaminase), (–)-epigallocatechinmo nogallate (an inhibitor of glutamate dehydrogenase) or L-buthionine sulfoximine (an inhibitor of glutathione synthesis). However, virus replication was rescued by the addition of multiple tricarboxylic acid cycle intermediates, ATP, or glutathione reduced ethyl ester. ATP and reduced glutathione/oxidized glutathione levels were increased in CPB cells infected with ISKNV, but were decreased in CPB cells cultured in glutamine-depleted medium. These results indicate ISKNV infection induces glutaminolysis to accommodate the biosynthetic and energy needs for its efficient virus replication.