TMT-based quantitative proteomics analysis reveals the attenuated replication mechanism of Newcastle disease virus caused by nuclear localization signal mutation in viral matrix protein.
TMT-based quantitative proteomics analysis reveals the attenuated replication mechanism of Newcastle disease virus caused by nuclear localization signal mutation in viral matrix protein.
复制标题
基于TMT的定量蛋白质组学分析揭示病毒基质蛋白核定位信号突变引起新城疫病毒复制减毒机制
DOI：
10.1080/21505594.2020.1770482
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发表时间：
2020-12
期刊：
影响因子：
5.2
通讯作者：
Ji X
中科院分区：
文献类型：
--
作者：
Duan Z;Yuan C;Han Y;Zhou L;Zhao J;Ruan Y;Chen J;Ni M;Ji X
摘要由内在核定位信号（NLS）介导的RNA病毒胞质蛋白的核定位在病毒成功复制中起着重要作用。我们曾报道过基质（M）蛋白的NLS突变明显减弱了纽卡斯尔病病毒（NDV）的复制和致病性，但其减弱复制的机制尚不清楚。在本研究中，我们发现M/NLS突变不仅破坏了M的核质运输特性，而且还损害了病毒RNA的合成和转录。利用基于TMT的定量蛋白质组学分析方法，对亲本NDV rSS 1GFP和携带M/NLS突变的突变NDV rSS 1GFP-M/NLSm感染BSR-T7/5细胞后的蛋白质组学结果进行分析，发现rSS 1GFP感染引起的差异表达蛋白质的数量和表达量显著高于rSS 1GFP-M/NLSm感染。进一步深入分析发现，M蛋白在细胞核内的优势积累抑制了宿主细胞的转录、RNA加工和修饰、蛋白质合成、转录后修饰和转运;当大部分M蛋白被限制在细胞核外时，这种抑制作用会减弱。更重要的是，我们发现M蛋白在细胞质中的功能以剂量依赖的方式抑制TIFA的表达，并通过下调TIFA/TRAF 6/NF-κ B介导的细胞因子的产生来促进NDV的复制。这是首次报道M蛋白参与NDV的免疫逃避。这些结果表明，NDV的复制与M蛋白的核质运输密切相关，这有助于我们对NDV M蛋白分子功能的理解。
ABSTRACT Nuclear localization of cytoplasmic RNA virus proteins mediated by intrinsic nuclear localization signal (NLS) plays essential roles in successful virus replication. We previously reported that NLS mutation in the matrix (M) protein obviously attenuates the replication and pathogenicity of Newcastle disease virus (NDV), but the attenuated replication mechanism remains unclear. In this study, we showed that M/NLS mutation not only disrupted M’s nucleocytoplasmic trafficking characteristic but also impaired viral RNA synthesis and transcription. Using TMT-based quantitative proteomics analysis of BSR-T7/5 cells infected with the parental NDV rSS1GFP and the mutant NDV rSS1GFP-M/NLSm harboring M/NLS mutation, we found that rSS1GFP infection stimulated much greater quantities and more expression changes of differentially expressed proteins involved in host cell transcription, ribosomal structure, posttranslational modification, and intracellular trafficking than rSS1GFP-M/NLSm infection. Further in-depth analysis revealed that the dominant nuclear accumulation of M protein inhibited host cell transcription, RNA processing and modification, protein synthesis, posttranscriptional modification and transport; and this kind of inhibition could be weakened when most of M protein was confined outside the nucleus. More importantly, we found that the function of M protein in the cytoplasm effected the inhibition of TIFA expression in a dose-dependent manner, and promoted NDV replication by down-regulating TIFA/TRAF6/NF-κB-mediated production of cytokines. It was the first report about the involvement of M protein in NDV immune evasion. Taken together, our findings demonstrate that NDV replication is closely related to the nucleocytoplasmic trafficking of M protein, which accelerates our understanding of the molecular functions of NDV M protein.
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影响因子：
3.7
作者：
Chutiwitoonchai N;Mano T;Kakisaka M;Sato H;Kondoh Y;Osada H;Kotani O;Yokoyama M;Sato H;Aida Y
通讯作者：
Aida Y
影响因子：
5.6
作者：
Guo, Haixia;Guo, Huihui;Zeng, Fanchang
通讯作者：
Zeng, Fanchang
影响因子：
3.7
作者：
Ahmed, M;Lyles, DS
通讯作者：
Lyles, DS
影响因子：
2.7
作者：
Duan, Zhiqiang;Song, Qingqing;Liu, Xiufan
通讯作者：
Liu, Xiufan
新城疫病毒基质蛋白的核定位通过影响病毒RNA合成和转录并抑制宿主细胞转录来促进病毒复制
影响因子：
4.4
作者：
Duan, Zhiqiang;Deng, Shanshan;Gao, Hongbo
通讯作者：
Gao, Hongbo