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草鱼肿瘤坏死因子受体相关蛋白1(CiTrap1)的蛋白质组学鉴定与表征:一种受草鱼呼肠孤病毒感染上调的抗凋亡因子

Proteomic identification and characterization of Ctenopharyngodon idella tumor necrosis factor receptor-associated protein 1 (CiTrap1): an anti-apoptosis factor upregulated by grass carp reovirus infection.

作者信息

Li Yan, Zhang Yanan, Wang Tu, Podok Patarida, Xu Dan, Lu Liqun

机构信息

Key Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, PR China.

Key Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, PR China.

出版信息

Fish Shellfish Immunol. 2015 Apr;43(2):449-59. doi: 10.1016/j.fsi.2015.01.031. Epub 2015 Feb 3.

Abstract

Human tumor necrosis factor receptor-associated protein 1 (Trap1) is a mitochondrial protein identical to heat shock protein 75 (HSP75) that plays an important role in protecting cells from oxidative stress and apoptosis. In this study, grass carp (Ctenopharyngodon idella) tumor necrosis factor receptor-associated protein 1 (designated as CiTrap1) was identified through two-dimensional electrophoresis (2-DE) analysis and its pattern of expression was investigated in grass carp kidney (CIK) cells infected with grass carp reovirus (GCRV). The full length cDNA of CiTrap1 contained an opening reading frame of 2157 bp that encoded a peptide of 718 amino acids. Phylogenetic analyses indicated that the CiTrap1 shared 87% identity with its homologue from zebrafish (Danio rerio). The transcriptional level of CiTrap1 in CIK cells was upregulated post virus infection as well as poly (I: C) stimulation. Following virus infection, grass carp PTEN-induced putative kinase 1 (PINK1) and Sorcin, whose coding proteins interact with Trap1 in human, were simultaneously upregulated with CiTrap1. Typical characteristics of apoptosis were observed in CIK cells infected with GCRV by DAPI staining, DNA ladder electrophoresis, TUNEL assay and Annexin Ⅴ labeling. RNAi-mediated silencing of CiTrap1 in CIK cells resulted in the increased rate of virus-induced apoptotic cells. The results of this study suggest that CiTrap1 is involved in the host's innate immune response to viral infection possibly through protecting infected cells from apoptosis.

摘要

人类肿瘤坏死因子受体相关蛋白1(Trap1)是一种与热休克蛋白75(HSP75)相同的线粒体蛋白,在保护细胞免受氧化应激和凋亡方面发挥着重要作用。在本研究中,通过二维电泳(2-DE)分析鉴定了草鱼(Ctenopharyngodon idella)肿瘤坏死因子受体相关蛋白1(命名为CiTrap1),并研究了其在感染草鱼呼肠孤病毒(GCRV)的草鱼肾(CIK)细胞中的表达模式。CiTrap1的全长cDNA包含一个2157 bp的开放阅读框,编码一个718个氨基酸的肽段。系统发育分析表明,CiTrap1与其斑马鱼(Danio rerio)同源物的同一性为87%。病毒感染以及聚(I:C)刺激后,CIK细胞中CiTrap1的转录水平上调。病毒感染后,草鱼PTEN诱导的假定激酶1(PINK1)和Sorcin(其编码蛋白在人类中与Trap1相互作用)与CiTrap1同时上调。通过DAPI染色、DNA梯状电泳、TUNEL检测和膜联蛋白Ⅴ标记,在感染GCRV的CIK细胞中观察到典型的凋亡特征。RNAi介导的CIK细胞中CiTrap1沉默导致病毒诱导的凋亡细胞率增加。本研究结果表明,CiTrap1可能通过保护受感染细胞免于凋亡而参与宿主对病毒感染的先天免疫反应。

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