A Proteomic Atlas of the African Swine Fever Virus Particle
A Proteomic Atlas of the African Swine Fever Virus Particle
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DOI：
10.1128/jvi.01293-18
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发表时间：
2018-12-01
影响因子：
5.4
通讯作者：
Andres, German
中科院分区：
文献类型：
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作者：
Alejo, Alf;Matamoros, Tania;Andres, German
非洲猪瘟病毒（ASFV）是一种大而复杂的DNA病毒，可引起高度致命的猪病，目前尚无疫苗。ASFV颗粒具有二十面体多层结构，含有多种多肽，其身份在很大程度上是未知的。在这里，我们通过质谱分析了高度纯化的细胞外ASFV颗粒的蛋白质组成，并进行免疫电子显微镜定位检测到的蛋白质。蛋白质组学分析鉴定了68种病毒蛋白，占基因组编码能力的39%。ASFV蛋白质组基本上包括所有以前描述的病毒粒子蛋白，有趣的是，44个新鉴定的病毒包装多肽，其中一半具有未知的功能。大部分病毒粒子蛋白致力于病毒结构，包括两个新鉴定的结构蛋白，p5和p8，它们分别来自核心多聚蛋白pp220和pp62。此外，病毒体含有参与病毒转录的酶和因子的完整补充，涉及DNA修复和蛋白质修饰的各种酶，以及与病毒进入和宿主防御逃避有关的一些蛋白质。最后，21个宿主蛋白，其中许多定位在细胞表面和相关的皮质肌动蛋白细胞骨架，可重复检测的ASFV颗粒。免疫电子显微镜强烈支持的建议，这些主机膜相关蛋白在病毒出芽期间在肌动蛋白依赖性膜突起招募。总之，这项研究的结果提供了一个全面的模型的ASFV架构，集成了组成和结构的信息。重要性非洲猪瘟病毒引起一种高度传染性和致命性的疾病，目前影响到许多国家的撒哈拉以南非洲，高加索，俄罗斯联邦和东欧，最近已蔓延到中国。尽管进行了广泛的研究，但仍然缺乏有效的疫苗或抗病毒策略，并且仍然存在许多关于感染周期的分子机制的基本问题。一个这样的差距涉及感染性病毒颗粒的组成和结构。在本报告所述的研究中，我们鉴定了组成病毒体的病毒和宿主蛋白，并确定或推断了其中许多蛋白的定位。这些信息大大增加了我们对传染性非洲猪瘟病毒颗粒的生物学和结构特征的了解，并将有助于指导未来的研究工作。
African swine fever virus (ASFV) is a large and complex DNA virus that causes a highly lethal swine disease for which there is no vaccine available. The ASFV particle, with an icosahedral multilayered structure, contains multiple polypeptides whose identity is largely unknown. Here, we analyzed by mass spectroscopy the protein composition of highly purified extracellular ASFV particles and performed immunoelectron microscopy to localize several of the detected proteins. The proteomic analysis identified 68 viral proteins, which account for 39% of the genome coding capacity. The ASFV proteome includes essentially all the previously described virion proteins and, interestingly, 44 newly identified virus-packaged polypeptides, half of which have an unknown function. A great proportion of the virion proteins are committed to the virus architecture, including two newly identified structural proteins, p5 and p8, which are derived from the core poly-proteins pp220 and pp62, respectively. In addition, the virion contains a full complement of enzymes and factors involved in viral transcription, various enzymes implicated in DNA repair and protein modification, and some proteins concerned with virus entry and host defense evasion. Finally, 21 host proteins, many of them localized at the cell surface and related to the cortical actin cytoskeleton, were reproducibly detected in the ASFV particle. Immunoelectron microscopy strongly supports the suggestion that these host membrane-associated proteins are recruited during virus budding at actin-dependent membrane protrusions. Altogether, the results of this study provide a comprehensive model of the ASFV architecture that integrates both compositional and structural information.IMPORTANCE African swine fever virus causes a highly contagious and lethal disease of swine that currently affects many countries of sub-Saharan Africa, the Caucasus, the Russian Federation, and Eastern Europe and has very recently spread to China. Despite extensive research, effective vaccines or antiviral strategies are still lacking, and many basic questions on the molecular mechanisms underlying the infective cycle remain. One such gap regards the composition and structure of the infectious virus particle. In the study described in this report, we identified the set of viral and host proteins that compose the virion and determined or inferred the localization of many of them. This information significantly increases our understanding of the biological and structural features of an infectious African swine fever virus particle and will help direct future research efforts.