Chang Jui-Shin, Chi Shau-Chi
Department of Life Science, National Taiwan University, Taipei 10617, Taiwan.
Department of Life Science, National Taiwan University, Taipei 10617, Taiwan.
Fish Shellfish Immunol. 2015 Oct;46(2):315-22. doi: 10.1016/j.fsi.2015.05.040. Epub 2015 Jun 4.
Nervous necrosis virus (NNV) is a non-enveloped virus with 2 segmented positive-sense single-stranded RNAs. NNV-induced mass mortality has occurred worldwide in many species of cultured marine fish and resulted in substantial economic losses. In our previous study, we cloned the gene of voltage-dependent anion selective channel protein 2 (GVDAC2), and the NNV RNA2 expression level decreased in GVDAC2-knockdown GF-1 cells 24 h after infection. Here, we investigated the role of GVDAC2 in the NNV infection in GF-1 cells. NNV infection did not considerably affect GVDAC2 gene expression. After performing immunostaining, we detected GVDAC2 at the mitochondria and GVDAC2 was colocalized with NNV-RNA-dependent RNA polymerase. However, these 2 proteins did not interact with each other in immunoprecipitation assay. The cellular ATP level in GVDAC2-downregulated cells was lower than that in control cells, and NNV-induced apoptosis was delayed in GVDAC2-siRNA-transfected cells. Therefore, we suggest that GVDAC2 is required for NNV infection for maintaining the cellular ATP level and has positive impact on virus-induced apoptosis.
神经坏死病毒（NNV）是一种无包膜病毒，具有两条分段的正链单链RNA。NNV引起的大规模死亡在全球范围内的许多养殖海水鱼类物种中都有发生，并造成了巨大的经济损失。在我们之前的研究中，我们克隆了电压依赖性阴离子选择性通道蛋白2（GVDAC2）的基因，并且在感染后24小时，GVDAC2基因敲低的GF-1细胞中NNV RNA2的表达水平下降。在此，我们研究了GVDAC2在GF-1细胞中NNV感染过程中的作用。NNV感染对GVDAC2基因表达没有显著影响。进行免疫染色后，我们在线粒体中检测到GVDAC2，并且GVDAC2与NNV RNA依赖性RNA聚合酶共定位。然而，在免疫沉淀试验中这两种蛋白质没有相互作用。GVDAC2下调细胞中的细胞ATP水平低于对照细胞，并且在GVDAC2-siRNA转染的细胞中NNV诱导的细胞凋亡延迟。因此，我们认为GVDAC2是NNV感染维持细胞ATP水平所必需的，并且对病毒诱导的细胞凋亡有积极影响。
