Capsid protein VP4 of human rhinovirus induces membrane permeability by the formation of a size-selective multimeric pore.
Capsid protein VP4 of human rhinovirus induces membrane permeability by the formation of a size-selective multimeric pore.
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DOI：
10.1371/journal.ppat.1004294
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发表时间：
2014-08
期刊：
影响因子：
6.7
通讯作者：
Tuthill TJ
中科院分区：
文献类型：
--
作者：
Panjwani A;Strauss M;Gold S;Wenham H;Jackson T;Chou JJ;Rowlands DJ;Stonehouse NJ;Hogle JM;Tuthill TJ
无包膜病毒必须将其病毒基因组递送穿过细胞膜，而没有膜融合的优势。对实现这一目标的机制仍然知之甚少。人类鼻病毒是普通感冒的常见病因，是小核糖核酸病毒家族的一种无包膜病毒，该家族包括脊髓灰质炎病毒和口蹄疫病毒等其他重要病原体。在小核糖核酸病毒进入细胞期间，小豆蔻酰化衣壳蛋白VP 4从病毒释放，与细胞膜相互作用，并参与将病毒RNA基因组递送到细胞质中以启动复制。在这项研究中，我们已经产生了重组C-末端组氨酸标记的人鼻病毒VP 4，并显示它可以诱导脂质体模型膜的膜通透性。葡聚糖尺寸排阻研究、化学交联和电子显微镜证实，VP 4形成多聚体膜孔，其通道尺寸与单链RNA基因组的转移一致。由重组VP 4诱导的膜渗透性受pH影响，并且与由感染性病毒体诱导的渗透性相当。这些发现提出了VP 4参与细胞进入的分子机制，并提供了一个模型系统，这将有助于探索VP 4作为小核糖核酸病毒家族的新的抗病毒靶点。人鼻病毒（HRV）是一种小核糖核酸病毒家族的无包膜病毒，每年造成数十亿美元的呼吸道感染（普通感冒）。仍然没有疫苗或许可的药物来预防或减少感染。小核糖核酸病毒家族的相关成员包括重要的病原体，如脊髓灰质炎病毒、肠道病毒71和口蹄疫病毒，对这些病毒也需要改进控制措施。病毒感染的一个基本步骤是通过细胞膜屏障传递病毒遗传物质。病毒如HIV和流感病毒被包裹在外膜中，该外膜可以与宿主细胞膜融合以允许病毒基因组渗透到细胞质中。然而，无包膜病毒如小核糖核酸病毒缺乏膜，这些病毒穿透膜的机制仍然知之甚少。衣壳蛋白VP 4先前已涉及小核糖核酸病毒基因组的递送。在这项研究中，我们证明了HRV VP 4与膜相互作用，使它们通过形成多聚体，大小选择性的膜孔与通过膜的病毒基因组的运输一致的属性渗透。VP 4的这种功能为该病毒家族提供了一种新的抗病毒靶点。
Non-enveloped viruses must deliver their viral genome across a cell membrane without the advantage of membrane fusion. The mechanisms used to achieve this remain poorly understood. Human rhinovirus, a frequent cause of the common cold, is a non-enveloped virus of the picornavirus family, which includes other significant pathogens such as poliovirus and foot-and-mouth disease virus. During picornavirus cell entry, the small myristoylated capsid protein VP4 is released from the virus, interacts with the cell membrane and is implicated in the delivery of the viral RNA genome into the cytoplasm to initiate replication. In this study, we have produced recombinant C-terminal histidine-tagged human rhinovirus VP4 and shown it can induce membrane permeability in liposome model membranes. Dextran size-exclusion studies, chemical crosslinking and electron microscopy demonstrated that VP4 forms a multimeric membrane pore, with a channel size consistent with transfer of the single-stranded RNA genome. The membrane permeability induced by recombinant VP4 was influenced by pH and was comparable to permeability induced by infectious virions. These findings present a molecular mechanism for the involvement of VP4 in cell entry and provide a model system which will facilitate exploration of VP4 as a novel antiviral target for the picornavirus family. Human rhinovirus (HRV) is a non-enveloped virus of the picornavirus family and is responsible for respiratory infections (common colds) costing billions of dollars ($) annually. There remains no vaccine or licensed drug to prevent or reduce infection. Related members of the picornavirus family include significant pathogens such as poliovirus, enterovirus 71 and foot-and-mouth disease virus, for which improved control measures are also required. A fundamental step in virus infection is the delivery of the viral genetic material through the barrier of the cellular membrane. Viruses such as HIV and influenza are enveloped in an outer membrane which can fuse with the host cell membrane to allow the viral genome to penetrate into the cytoplasm. However, non-enveloped viruses such as picornaviruses lack a membrane and the mechanisms for penetration of the membrane by these viruses remain poorly understood. The capsid protein VP4 has previously been implicated in the delivery of the picornavirus genome. In this study we demonstrate that HRV VP4 interacts with membranes to make them permeable by the formation of multimeric, size-selective membrane pores with properties consistent with the transport of viral genome through the membrane. This function of VP4 provides a novel antiviral target for this family of viruses.
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