Analysis of virion associated host proteins in vesicular stomatitis virus using a proteomics approach.
Analysis of virion associated host proteins in vesicular stomatitis virus using a proteomics approach.
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DOI：
10.1186/1743-422x-6-166
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发表时间：
2009-10-12
期刊：
影响因子：
4.8
通讯作者：
Grdzelishvili VZ
中科院分区：
文献类型：
--
作者：
Moerdyk-Schauwecker M;Hwang SI;Grdzelishvili VZ
水泡性口炎病毒（VSV）是典型的弹状病毒，也是单负病毒目中研究得最好的成员。现在有令人信服的证据表明，从感染细胞释放的包膜病毒粒子携带许多宿主（细胞）蛋白，其中一些可能在病毒复制中发挥重要作用。尽管先前已经显示几种细胞蛋白质被掺入VSV病毒体中，但是还没有进行系统的研究来揭示VSV病毒体或单负性病毒目的任何其他成员的宿主蛋白质组成。在这里，我们使用了蛋白质组学的方法来确定纯化的VSV病毒粒子内的细胞蛋白，从而创建一个病毒/宿主相互作用的一个阶段的“快照”，可以指导未来的实验，旨在了解病毒-细胞相互作用的分子机制。使用质谱法分析来自人、小鼠和仓鼠来源的三种不同细胞系的VSV病毒体的高度纯化制剂中细胞蛋白的存在。我们已经成功地证实了VSV病毒粒子内存在几种以前鉴定的细胞蛋白，并鉴定了一些可能也存在于病毒粒子内的其他蛋白。总共鉴定了64种细胞蛋白，其中9种在多种制剂中发现。使用免疫印迹和蛋白酶K保护试验的组合来验证病毒体中存在这些蛋白质中的几种（整合素β1、热休克蛋白90 kDa、热休克同源71 kDa蛋白、膜联蛋白2、延伸因子1a）。据我们所知，这是第一次系统研究VSV或单负病毒目任何其他成员的病毒粒子的宿主蛋白质组成。需要进一步的实验来确定哪些鉴定的蛋白质与VSV具有相互作用，以及这些相互作用对于VSV复制是否是有益的、中性的或抗病毒的。鉴定对病毒有益的宿主蛋白-病毒相互作用将是特别令人兴奋的，因为它们可以提供通过控制宿主组分来对抗病毒感染的新方法。
Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and the best studied member of the order Mononegavirales. There is now compelling evidence that enveloped virions released from infected cells carry numerous host (cellular) proteins some of which may play an important role in viral replication. Although several cellular proteins have been previously shown to be incorporated into VSV virions, no systematic study has been done to reveal the host protein composition for virions of VSV or any other member of Mononegavirales. Here we used a proteomics approach to identify cellular proteins within purified VSV virions, thereby creating a "snapshot" of one stage of virus/host interaction that can guide future experiments aimed at understanding molecular mechanisms of virus-cell interactions. Highly purified preparations of VSV virions from three different cell lines of human, mouse and hamster origin were analyzed for the presence of cellular proteins using mass spectrometry. We have successfully confirmed the presence of several previously-identified cellular proteins within VSV virions and identified a number of additional proteins likely to also be present within the virions. In total, sixty-four cellular proteins were identified, of which nine were found in multiple preparations. A combination of immunoblotting and proteinase K protection assay was used to verify the presence of several of these proteins (integrin β1, heat shock protein 90 kDa, heat shock cognate 71 kDa protein, annexin 2, elongation factor 1a) within the virions. This is, to our knowledge, the first systematic study of the host protein composition for virions of VSV or any other member of the order Mononegavirales. Future experiments are needed to determine which of the identified proteins have an interaction with VSV and whether these interactions are beneficial, neutral or antiviral with respect to VSV replication. Identification of host proteins-virus interactions beneficial for virus would be particularly exciting as they can provide new ways to combat viral infections via control of host components.