Determination of the cleavage site of enterovirus 71 VP0 and the effect of this cleavage on viral infectivity and assembly
Determination of the cleavage site of enterovirus 71 VP0 and the effect of this cleavage on viral infectivity and assembly
复制标题
确定肠道病毒 71 VP0 的裂解位点以及该裂解对病毒感染性和组装的影响
DOI：
10.1016/j.micpath.2019.103568
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发表时间：
2019-09-01
影响因子：
3.8
通讯作者：
Jiang, Chunlai
中科院分区：
文献类型：
--
作者：
Cao, Jiaming;Liu, Hongtao;Jiang, Chunlai
手足口病(HFMD)是一个主要的公共卫生问题，尤其是在婴幼儿中。手足口病的主要病原是肠道病毒71型(EV71)，其衣壳组装机制包括衣壳蛋白的加工已被广泛研究。然而，它的一些机制仍不清楚，如VP0裂解。本研究旨在确定EV71VP0衣壳蛋白的裂解位点，并阐明EV71VP0裂解对病毒感染性和装配的影响。质谱分析表明，EV71 VP0的裂解位点位于残基Lys69和Ser70之间。为了分析任一个残基对切割的重要性，我们分别设计了Lys69、Ser70和双突变的单突变，并对这些基因组进行了封装。结果表明，Ser70对VP0的裂解和EV71的感染性更为重要。此外，还利用EV71蛋白水解酶2A和3C的外源表达来验证它们是否在VP0切割中起作用。分析还表明，他们中没有一个人参与这一进程。本研究对EV71衣壳成熟的机制提供了新的见解，可能成为提高EV71疫苗生产效率和免疫原性的潜在靶点。
Hand, foot, and mouth disease (HFMD) is a major public health concern, especially among infants and young children. The primary pathogen of HFMD is enterovirus 71 (EV71), whose capsid assembly mechanism including capsid protein processing has been widely studied. However, some of its mechanisms remain unclear, such as the VP0 cleavage. This study aimed to identify the cleavage site of the EV71 VP0 capsid protein and to elucidate the effects of EV71 VP0 cleavage on viral infectivity and assembly. A mass spectrometry analysis indicated that the cleavage site of EV71 VP0 is located between residues Lys69 and Ser70. To analyze the importance of either residue to cleavage, we designed single mutations of Lys69, Ser70 and double mutations respectively and implemented these genomes to encapsulation. The results indicated that Ser70 is more important for VP0 cleavage and EV71 infectivity. In addition, exogenous expression of EV71 protease 2A and 3C was used to verify whether they play roles in VP0 cleavage. Analyses also showed that none of them participate in this process. This study provides novel insights into the mechanisms of EV71 capsid maturation, which may be a potential target to improve the productivity and immunogenicity of EV71 vaccines.