Identification of Interactions between Sindbis Virus Capsid Protein and Cytoplasmic vRNA as Novel Virulence Determinants.
Identification of Interactions between Sindbis Virus Capsid Protein and Cytoplasmic vRNA as Novel Virulence Determinants.
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DOI：
10.1371/journal.ppat.1006473
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发表时间：
2017-06
期刊：
影响因子：
6.7
通讯作者：
Hardy RW
中科院分区：
文献类型：
--
作者：
Sokoloski KJ;Nease LM;May NA;Gebhart NN;Jones CE;Morrison TE;Hardy RW
甲病毒是节肢动物传播的病毒，在全球范围内对公共卫生构成重大威胁。虽然由货物核酸和病毒衣壳蛋白组成的甲病毒核衣壳核心的形成是感染的重要分子过程，但两种伴侣之间的精确相互作用尚不明确。CLIP-seq方法用于筛选辛德毕斯病毒（SINV）（一种模型甲病毒）的病毒衣壳蛋白与基因组RNA之间相互作用的候选位点。本报告中提供的数据表明，SINV衣壳蛋白与感染细胞的细胞质中的特异性病毒RNA序列结合，但其与成熟细胞外病毒颗粒中的基因组RNA的相互作用在核苷酸序列方面基本上是非特异性的。细胞质病毒RNA-衣壳相互作用位点的突变分析揭示了衣壳结合在感染早期的功能作用。相互作用位点突变体表现出病毒生长动力学下降，但是，这种缺陷是不是减少颗粒生产的功能。相反，细胞质衣壳-RNA相互作用位点的突变负面影响了进入的病毒基因组RNA的功能能力，导致感染性降低。此外，细胞质衣壳相互作用位点突变体在嗜神经性甲病毒感染的鼠模型中减毒。总的来说，这项研究的结果表明，所确定的病毒衣壳蛋白和基因组RNA的细胞质相互作用，而不是必不可少的颗粒形成，是必要的基因组RNA功能在感染早期。衣壳蛋白在甲病毒复制周期中的这种先前未被认识到的作用也构成了一种新的毒力决定因素。甲病毒可在受感染个体中引起重大疾病;然而，我们对使感染和促成疾病发展的分子相互作用的理解是有限的。本手稿中详细描述的工作描述了病毒RNA结合蛋白衣壳与病毒基因组RNA的相互作用。重要的是，这些相互作用被发现在基因组上的特定位点，但不是病毒组装所必需的。在小鼠感染模型中，衣壳/ RNA相互作用位点的突变降低了病毒的复制和疾病的严重程度。总之，这些发现确定了以前未发现的疾病严重程度的决定因素，并为开发新疫苗提供了潜在的基础。
Alphaviruses are arthropod-borne viruses that represent a significant threat to public health at a global level. While the formation of alphaviral nucleocapsid cores, consisting of cargo nucleic acid and the viral capsid protein, is an essential molecular process of infection, the precise interactions between the two partners are ill-defined. A CLIP-seq approach was used to screen for candidate sites of interaction between the viral Capsid protein and genomic RNA of Sindbis virus (SINV), a model alphavirus. The data presented in this report indicates that the SINV capsid protein binds to specific viral RNA sequences in the cytoplasm of infected cells, but its interaction with genomic RNA in mature extracellular viral particles is largely non-specific in terms of nucleotide sequence. Mutational analyses of the cytoplasmic viral RNA-capsid interaction sites revealed a functional role for capsid binding early in infection. Interaction site mutants exhibited decreased viral growth kinetics; however, this defect was not a function of decreased particle production. Rather mutation of the cytoplasmic capsid-RNA interaction sites negatively affected the functional capacity of the incoming viral genomic RNAs leading to decreased infectivity. Furthermore, cytoplasmic capsid interaction site mutants are attenuated in a murine model of neurotropic alphavirus infection. Collectively, the findings of this study indicate that the identified cytoplasmic interactions of the viral capsid protein and genomic RNA, while not essential for particle formation, are necessary for genomic RNA function early during infection. This previously unappreciated role of capsid protein during the alphaviral replication cycle also constitutes a novel virulence determinant. Alphaviruses can cause significant disease in infected individuals; however, our understanding of the molecular interactions that enable infection and contribute to the development of disease is limited. The work detailed in this manuscript characterizes the interaction of a viral RNA-binding protein, Capsid, with the viral genomic RNA. Importantly, these interactions were found to be at specific sites on the genome but not essential for virus assembly. Mutation of the capsid / RNA interaction sites decreased the replication of the virus and the severity of disease in a mouse model of infection. Taken together, these findings identify a previously undiscovered determinant of disease severity, and provide a potential basis for the development of new vaccines.
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影响因子：
3.7
作者：
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