生物学杂志

生物学杂志 ›› 2024, Vol. 41 ›› Issue (3): 92-.doi: 10.3969/j.issn.2095-1736.2024.03.092

• 研究报告 • 上一篇    下一篇

利用邻近标记-质谱联用技术筛选埃博拉病毒相关宿主因子

张　迅1， 柏　宇1， 刘海楠2， 刘　萱2， 曹　诚2   

  1. 1. 安徽大学 物质科学与信息技术研究院， 合肥 230601； 2. 军事科学院 军事医学研究院， 北京 100850
  • 出版日期:2024-06-18 发布日期:2024-06-17
  • 通讯作者: 刘萱，研究员,从事病原微生物与宿主互作研究，E-mail:liux931932@163.com
  • 作者简介:张迅，硕士研究生,从事病原微生物与宿主互作研究，E-mail:zhangxun2453@163.com
  • 基金资助:
    中国科学院武汉国家生物安全实验室高端用户培育项目（2022ACCP-MS04）； 国家科技重大专项（2018ZX09711003-005-005）

Utilization of proximity labeling-mass spectrometry technique to identify Ebola virus-associated host factor in Human

ZHANG Xun1, BAI Yu1, LIU Hainan2, LIU Xuan2, CAO Cheng2   

  1. 1. Institute of Physical Science and Information Technology, Anhui University, Hefei 230601, China;
    2. Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
  • Online:2024-06-18 Published:2024-06-17

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摘要： 构建VP35与生物素连接酶TurboID融合蛋白，利用埃博拉病毒最小基因组系统（Ebola virus minigenome system， EBOV MG），在细胞中模拟病毒的生命周期和病毒包涵体（virus inlusion body，VIB）的形成过程，通过添加外源生物素，标记VP35相互作用蛋白。定量质谱丰度差异分析筛选出537个潜在的VP35互作蛋白，Gene Ontology（GO）分析发现其中包括大量与RNA结合相关蛋白。从中选取EIF4B和ZNF598进行初步验证，二者均与VP35存在相互作用，且敲低上述基因可显著抑制EBOV 转录复制病毒样颗粒（trVLP）的复制效率。结果证实，邻近标记-质谱联用技术可以用于病毒-宿主相互作用蛋白挖掘，为病毒致病机制研究及抗病毒靶点挖掘提供重要手段。

关键词: 邻近标记, 埃博拉病毒, 病毒包涵体, TurboID, VP35

Abstract: The fusion protein of VP35 and biotin ligase TurboID was generated, and the Ebola virus minigenome system (EBOV MG) was utilized to simulate the Ebola virus lifecycle and formation of virus inclusion bodies (VIB) in cells. The introduction of exogenous biotin enabled the labeling of proteins to interact with VP35. Among all proximity labeling (PL) labeled proteins, 537 potential VP35-associated host proteins were identified by differential abundance analysis after quantitative mass spectrometry detection. Gene Ontology (GO) analysis found that many enriched proteins were involved in RNA binding related functions. Subsequently, the association of EBOV VP35 with RNA binding proteins EIF4B and ZNF598 was confirmed. Interruption of EIF4B and ZNF598 expression significantly inhibited EBOV trVLP replication. The study highlighted the effectiveness of PL-quantitative mass spectrometry in identifying virus-host interaction proteins, providing a valuable tool for investigating viral pathogenesis and identifying potential antiviral targets.

Key words: proximity labeling, EBOV, viral inclusion bodies, TurboID, VP35

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