Interaction between a Unique Minor Protein and a Major Capsid Protein of Bluetongue Virus Controls Virus Infectivity.
Interaction between a Unique Minor Protein and a Major Capsid Protein of Bluetongue Virus Controls Virus Infectivity.
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独特的小蛋白质与蓝肠病毒的主要衣壳蛋白之间的相互作用可控制病毒感染性。
DOI：
10.1128/jvi.01784-17
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发表时间：
2018-02-01
影响因子：
5.4
通讯作者：
Roy P
中科院分区：
文献类型：
--
作者：
Matsuo E;Yamazaki K;Tsuruta H;Roy P
在呼肠孤病毒科的双链RNA病毒中，只有环状病毒属的成员在其颗粒内具有独特的结构蛋白，称为VP 6。蓝舌病病毒（Bluetongue virus，BTV）是一种重要的家畜病原病毒，是环状病毒的原型。BTV VP 6是一种依赖于ATP的RNA解旋酶，是病毒复制所必需的。在本报告所述的研究中，我们研究了VP 6如何被募集到病毒衣壳，以及形成病毒衣壳核心内层的BTV结构蛋白VP 3是否参与VP 6募集。我们首先证明了VP 6与VP 3相互作用，并在衣壳组装过程中与VP 3共定位。一系列的VP 6突变体，然后产生，并结合免疫沉淀和尺寸排阻色谱分析，我们证明，VP 6直接与VP 3通过一个特定区域的C-末端部分的VP 6。最后，利用我们的反向遗传学系统，将突变的VP 6蛋白引入BTV基因组中，并在活细胞系统中显示VP 6和VP 3之间的相互作用。我们证明，BTV株具有突变的VP 6是复制缺陷的野生型BSR细胞，并未能招募病毒复制酶复合物到病毒颗粒的核心。总之，这些数据表明，VP 3和VP 6之间的相互作用可能是重要的病毒基因组的包装和颗粒形成的早期阶段。重要性环状病毒蓝舌病病毒（BTV）是牲畜蓝舌病的病原体，通常对畜牧业造成重大的经济和农业影响。在本报告描述的研究中，我们确定了独特的环状病毒衣壳蛋白VP 6的重要区域和残基，这些区域和残基负责其与其他BTV蛋白的相互作用以及随后招募到病毒颗粒中。这些相互作用的性质和机制表明，VP 6在BTV基因组包装到病毒颗粒中具有关键作用。因此，这是一个非常重要的发现，因为这种对BTV组装的新理解可以用于设计针对蓝舌病的新型疫苗和抗病毒药。
Among the Reoviridae family of double-stranded RNA viruses, only members of the Orbivirus genus possess a unique structural protein, termed VP6, within their particles. Bluetongue virus (BTV), an important livestock pathogen, is the prototype Orbivirus. BTV VP6 is an ATP-dependent RNA helicase, and it is indispensable for virus replication. In the study described in this report, we investigated how VP6 might be recruited to the virus capsid and whether the BTV structural protein VP3, which forms the internal layer of the virus capsid core, is involved in VP6 recruitment. We first demonstrated that VP6 interacts with VP3 and colocalizes with VP3 during capsid assembly. A series of VP6 mutants was then generated, and in combination with immunoprecipitation and size exclusion chromatographic analyses, we demonstrated that VP6 directly interacts with VP3 via a specific region of the C-terminal portion of VP6. Finally, using our reverse genetics system, mutant VP6 proteins were introduced into the BTV genome and interactions between VP6 and VP3 were shown in a live cell system. We demonstrate that BTV strains possessing a mutant VP6 are replication deficient in wild-type BSR cells and fail to recruit the viral replicase complex into the virus particle core. Taken together, these data suggest that the interaction between VP3 and VP6 could be important in the packaging of the viral genome and early stages of particle formation. IMPORTANCE The orbivirus bluetongue virus (BTV) is the causative agent of bluetongue disease of livestock, often causing significant economic and agricultural impacts in the livestock industry. In the study described in this report, we identified the essential region and residues of the unique orbivirus capsid protein VP6 which are responsible for its interaction with other BTV proteins and its subsequent recruitment into the virus particle. The nature and mechanism of these interactions suggest that VP6 has a key role in packaging of the BTV genome into the virus particle. As such, this is a highly significant finding, as this new understanding of BTV assembly could be exploited to design novel vaccines and antivirals against bluetongue disease.