Comparative Proteomic Analysis of Wild-Type and SAP Domain Mutant Foot-and-Mouth Disease Virus-Infected Porcine Cells Identifies the Ubiquitin-Activating Enzyme UBE1 Required for Virus Replication.
Comparative Proteomic Analysis of Wild-Type and SAP Domain Mutant Foot-and-Mouth Disease Virus-Infected Porcine Cells Identifies the Ubiquitin-Activating Enzyme UBE1 Required for Virus Replication.
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DOI：
10.1021/acs.jproteome.5b00310
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发表时间：
2015-09
影响因子：
4.4
通讯作者：
Zixiang Zhu;Fan Yang;Keshan Zhang;W. Cao;Ye Jin;Guoqing Wang;R. Mao;Dan Li;Jian-hong Guo;Xiang-tao Liu;Haixue Zheng
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文献类型：
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作者：
Zixiang Zhu;Fan Yang;Keshan Zhang;W. Cao;Ye Jin;Guoqing Wang;R. Mao;Dan Li;Jian-hong Guo;Xiang-tao Liu;Haixue Zheng
口蹄疫病毒的前导蛋白L(Pro)通过调节多种宿主蛋白的活性来促进病毒的复制和致病。L(Pro)有一个保守的蛋白结构域SAP，被认为可以破坏干扰素的产生，从而阻断抗病毒反应。然而，除了阻断干扰素的产生外，SAP结构域在口蹄疫病毒感染宿主细胞中的作用尚不清楚。因此，我们通过高通量定量蛋白质组学方法[相对和绝对定量等压标签(ITRAQ)结合高效液相色谱/电喷雾电离串联质谱仪]鉴定了与L(Pro)SAP结构域相关的宿主蛋白质。比较感染Ra/FMDVSK6mSAP和Ra/FMDVSK6细胞中差异表达的蛋白质，发现45个下调的蛋白质和32个上调的蛋白质，这些蛋白质主要与代谢、核糖体、剪接体和泛素-蛋白酶体途径有关。这些结果还表明，SAP结构域除了具有潜在的激活信号转导和转录激活因子(PIAS)功能的蛋白抑制因子外，还具有与SAF-A/B类似的功能。进一步分析了其中一个已鉴定的蛋白UBe1，发现它与L(Pro)的SAP结构域有一个新的功能。UBE1的过表达促进了口蹄疫病毒的复制，而UBE1的敲除则降低了口蹄疫病毒的复制。这表明口蹄疫病毒操纵UBE1以增加病毒复制，SAP结构域参与了这一过程。
Leader protein (L(pro)) of foot-and-mouth disease virus (FMDV) manipulates the activities of several host proteins to promote viral replication and pathogenicity. L(pro) has a conserved protein domain SAP that is suggested to subvert interferon (IFN) production to block antiviral responses. However, apart from blocking IFN production, the roles of the SAP domain during FMDV infection in host cells remain unknown. Therefore, we identified host proteins associated with the SAP domain of L(pro) by a high-throughput quantitative proteomic approach [isobaric tags for relative and absolute quantitation (iTRAQ) in conjunction with liquid chromatography/electrospray ionization tandem mass spectrometry]. Comparison of the differentially regulated proteins in rA/FMDVΔmSAP- versus rA/FMDV-infected SK6 cells revealed 45 down-regulated and 32 up-regulated proteins that were mostly associated with metabolic, ribosome, spliceosome, and ubiquitin-proteasome pathways. The results also imply that the SAP domain has a function similar to SAF-A/B besides its potential protein inhibitor of activated signal transducer and activator of transcription (PIAS) function. One of the identified proteins UBE1 was further analyzed and displayed a novel role for the SAP domain of L(pro). Overexpression of UBE1 enhanced the replication of FMDV, and knockdown of UBE1 decreased FMDV replication. This shows that FMDV manipulates UBE1 for increased viral replication, and the SAP domain was involved in this process.