Proteomic analysis of cellular protein expression profiles in response to grass carp reovirus infection
Proteomic analysis of cellular protein expression profiles in response to grass carp reovirus infection
复制标题
草鱼呼肠孤病毒感染细胞蛋白表达谱的蛋白质组学分析
DOI：
10.1016/j.fsi.2015.03.010
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发表时间：
2015
影响因子：
4.7
通讯作者：
Lu Liqun
中科院分区：
文献类型：
--
作者：
Xu Dan;Song Lang;Wang Hao;Xu Xiaoyan;Wang Tu;Lu Liqun
草鱼出血症是由草鱼呼肠孤病毒(GCRV)引起的一种严重危害草鱼养殖的疾病。为了更好地了解GCRV感染后的分子反应，采用双向电泳(2-DE)和基质辅助激光解吸/电离串联质谱仪对CIK细胞进行了研究。比较模拟感染CIK细胞和感染GCRV的CIK细胞的差异表达蛋白质。共鉴定出23个差异表达点(22个上调点和1个下调点)，包括细胞骨架蛋白、大分子生物合成相关蛋白、应激反应蛋白、信号转导蛋白、能量代谢相关蛋白和泛素蛋白酶体途径相关蛋白。此外，通过实时定量逆转录聚合酶链式反应对差异表达蛋白中的10个相应基因进行了定量，以检测它们的转录图谱。草鱼细胞应激颗粒途径的T细胞内抗原1(TIA1)和Ras-GTP酶激活SH3区结合蛋白1(G3BP1)。在GCRV感染过程中，Idella(命名为CiTIA1和CiG3BP1)分别上调和下调。CiTIA1基因全长2753hp，开放阅读框(ORF)1155bp，编码385个氨基酸。CiG3BP1全长2271bp，编码485个氨基酸的开放阅读框架(ORF)。系统发育分析表明，CiTIA1和CiG3BP1的完整ORF与斑马鱼和哺乳动物的同源物非常相似。Western blotting验证了细胞蛋白CiTIA1和CiG3BP1对GCRV的应答，表明GCRV应解开TIA1的聚集和应激颗粒的形成。这项研究为蛋白质组和细胞应激颗粒途径对GCRV感染的反应提供了有用的信息，有助于我们对病毒发病机制的理解。
Grass carp (Ctenopharyngodon idella) hemorrhagic disease, caused by grass carp reovirus (GCRV), is emerging as a serious problem in grass carp aquaculture. To better understand the molecular responses to GCRV infection, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization tandem mass spectroscopy were performed to investigate altered proteins inC. idellakidney (CIK) cells. Differentially expressed proteins in mock infected CIK cells and GCRV-infected CIK cells were compared. Twenty-three differentially expressed spots were identified (22 upregulated spots and 1 downregulated spot), which included cytoskeleton proteins, macromolecular biosynthesis-associated proteins, stress response proteins, signal transduction proteins, energy metabolism-associated proteins and ubiquitin proteasome pathway-associated proteins. Moreover, 10 of the corresponding genes of the differentially expressed proteins were quantified by real-time reverse transcription polymerase chain reaction to examine their transcriptional profiles. The T cell internal antigen 1 (TIA1) and Ras-GTPase-activating SH3-domain-binding protein1 (G3BP1) of the cellular stress granule pathway from grass carpC. idella(designated asCiTIA1 andCiG3BP1) were upregulated and downregulated during GCRV infection, respectively. The full-length cDNA ofCiTIA1 was 2753 bp, with an open reading frame (ORF) of 1155bp, which encodes a putative 385-amino acid protein. The 2271bp full-length cDNA ofCiG3BP1 comprised an ORF of 1455bp that encodes a putative 485-amino acid protein. Phylogenetic analysis revealed that the complete ORFs ofCiTIA1 andCiG3BP1 were very similar to zebrafish and well-characterized mammalian homologs. The expressions of the cellular proteinsCiTIA1 andCiG3BP1 in response to GCRV were validated by western blotting, which indicated that the GCRV should unlink TIA1 aggregation and stress granule formation. This study provides useful information on the proteomic and cellular stress granule pathway's responses to GCRV infection, which adds to our understanding of viral pathogenesis.