Proteomic analysis of chicken embryo fibroblast cells infected with recombinant H5N1 avian influenza viruses with and without NS1 eIF4GI binding domain.
Proteomic analysis of chicken embryo fibroblast cells infected with recombinant H5N1 avian influenza viruses with and without NS1 eIF4GI binding domain.
复制标题
感染带有和不带有 NS1 eIF4GI 结合域的重组 H5N1 禽流感病毒的鸡胚成纤维细胞的蛋白质组学分析
DOI：
10.18632/oncotarget.23615
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发表时间：
2018-02-02
期刊：
影响因子：
--
通讯作者：
Jin M
中科院分区：
文献类型：
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作者：
Guo K;Lin X;Li Y;Qian W;Zou Z;Chen H;Zhou H;Jin M
非结构蛋白1（NS 1）是流感病毒的一个重要毒力因子，在流感病毒的复制过程中起着重要的调控作用。缺失NS 1的eIF 4GI结合结构域的重组H5 N1病毒（rNS 1-SD 30）表现出比具有完整eIF 4GI结合结构域的H5 N1病毒（rNS 1-wt）显著更低的致病性。为了进一步研究这一现象，我们进行了比较蛋白质组学分析，以分析感染rNS 1-wt和rNS 1-SD 30病毒的鸡胚成纤维细胞（CEF）中的宿主蛋白。总共有81个差异表达（DE）蛋白在感染后12，24和36小时确定。这些蛋白质主要参与细胞骨架、凋亡和应激反应、转录调节、运输和代谢过程、mRNA加工和剪接以及细胞信号转导。DE蛋白的过表达表明，ANXA 7抑制rNS 1-SD 30的繁殖，但不抑制rNS 1-wt病毒的繁殖。此外，ALDH 7A 1、ANXA 7和DCTN 2强烈增强鸡MDA 5（chMDA 5）诱导的IFN-β启动子活性，并且在ANXA 7的情况下，还增强rNS 1-SD 30病毒株诱导的IFN-β启动子活性。NS 1-wt共转染抑制了由chMDA 5诱导的ANXA 7介导的IFN-β启动子活性的增加。这些发现强调了NS 1 eIF 4GI结合结构域在H5 N1致病性中的作用，并可能有助于设计抗病毒策略以降低与该病原体相关的高发病率和死亡率。
Non-structural 1 (NS1) protein is a key virulence factor that regulates replication of influenza virus. A recombinant H5N1 virus lacking the eIF4GI-binding domain of NS1 (rNS1-SD30) exhibits significantly lower pathogenicity than H5N1 virus with an intact eIF4GI-binding domain (rNS1-wt). To further investigate this phenomenon, we performed comparative proteomics analyses to profile host proteins in chicken embryo fibroblasts (CEFs) infected with rNS1-wt and rNS1-SD30 viruses. In total, 81 differentially expressed (DE) proteins were identified at 12, 24, and 36 h post-infection. These proteins are mainly involved in the cytoskeletal, apoptotic and stress responses, transcription regulation, transport and metabolic processes, mRNA processing and splicing, and cellular signal transduction. Overexpression of DE proteins revealed that ANXA7 suppresses propagation of rNS1-SD30, but not rNS1-wt viruses. Moreover, ALDH7A1, ANXA7, and DCTN2 strongly enhanced IFN-β promoter activity induced by chicken MDA5 (chMDA5), and in the case of ANXA7, also by the rNS1-SD30 viral strain. NS1-wt co-transfection suppressed the ANXA7-mediated increase in IFN-β promoter activity induced by chMDA5. These findings highlight the role of NS1 eIF4GI binding domain in H5N1 pathogenicity, and may contribute to the design of antiviral strategies to reduce the high morbidity and mortality associated with this pathogen.