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采用蛋白质组学方法分析水疱性口炎病毒中的病毒相关宿主蛋白。

Analysis of virion associated host proteins in vesicular stomatitis virus using a proteomics approach.

机构信息

Department of Biology, University of North Carolina at Charlotte, Charlotte, NC 28223, USA.

出版信息

Virol J. 2009 Oct 12;6:166. doi: 10.1186/1743-422X-6-166.

Abstract

BACKGROUND

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.

RESULTS

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 beta1, heat shock protein 90 kDa, heat shock cognate 71 kDa protein, annexin 2, elongation factor 1a) within the virions.

CONCLUSION

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.

摘要

背景

水疱性口炎病毒(VSV)是原型弹状病毒,也是单负链 RNA 病毒目(Mononegavirales)中研究得最好的成员。现在有确凿的证据表明,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。尽管以前已经表明几种细胞蛋白被掺入 VSV 病毒粒子中,但尚未进行系统研究以揭示 VSV 或任何其他单负链 RNA 病毒成员的病毒粒子的宿主蛋白组成。

结果

在这里,我们使用蛋白质组学方法来鉴定纯化的 VSV 病毒粒子中的细胞蛋白,从而创建了病毒/宿主相互作用的一个“快照”,可以指导未来旨在理解病毒-细胞相互作用分子机制的实验。使用质谱法分析了来自人、鼠和仓鼠三种不同细胞系的高度纯化的 VSV 病毒粒子制剂中细胞蛋白的存在。我们已经成功地证实了几种先前鉴定的细胞蛋白存在于 VSV 病毒粒子中,并鉴定了一些可能也存在于病毒粒子中的其他蛋白。总共鉴定了 64 种细胞蛋白,其中 9 种存在于多个制剂中。免疫印迹和蛋白水解酶保护测定的组合用于验证这些蛋白中的几种(整合素β 1、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)在病毒粒子中的存在。

结论

这是迄今为止对 VSV 或单负链 RNA 病毒目中任何其他成员的病毒粒子的宿主蛋白组成的首次系统研究。未来的实验需要确定鉴定出的蛋白中有哪些与 VSV 相互作用,以及这些相互作用对 VSV 复制是有益、中性还是抗病毒。鉴定对病毒有益的宿主蛋白-病毒相互作用将特别令人兴奋,因为它们可以通过控制宿主成分提供新的方法来对抗病毒感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e635/2770056/57b1f6edb980/1743-422X-6-166-1.jpg

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