Identification of cellular proteins interacting with PEDV M protein through APEX2 labeling

文献类型： 外文期刊

第一作者： Dong, Shijuan

作者： Dong, Shijuan;Wang, Ruiyang;Yu, Ruisong;Chen, Bingqing;Si, Fusheng;Xie, Chunfang;Li, Zhen;Dong, Shijuan;Chen, Bingqing;Si, Fusheng;Li, Zhen;Li, Zhen

作者机构：

关键词： PEDV; Membrane protein; Cellular proteins; APEX2; Interaction

期刊名称：JOURNAL OF PROTEOMICS （ 影响因子：3.509； 五年影响因子：3.637 ）

ISSN： 1874-3919

年卷期： 2021 年 240 卷

页码：

收录情况： SCI

摘要： Membrane (M) proteins of coronaviruses are the most abundant component of the virus envelope and play crucial roles in virus assembly, virus budding and the regulation of host immunity. To understand more about these functions in the context of PEDV M protein, forty host cell proteins interacting with the M protein were identified in the present study by exploiting the proximity-labeling enzyme APEX2 (a mutant soybean ascorbate peroxidase). Bioinformatic analysis showed that the identified host cell proteins were related to fifty-four signal pathways and a wide diversity of biological processes. Interaction between M and five of the identified proteins (RIG-I, PPID, NHE-RF1, S100A11, CLDN4) was confirmed by co-immunoprecipitation (Co-IP). In addition, knockdown of PPID and S100A11 genes by siRNA significantly improved virus production, indicating that the proteins encoded by the two genes were interfering with or down-regulating virus replication in infected cells. Identification of the host cell proteins accomplished in this study provides new information about the mechanisms underlying PEDV replication and immune evasion. Significance: PEDV M protein is an essential structural protein implicated in viral infection, replication and assembly although the precise mechanisms underlying these functions remain enigmatic. In this study, we have identified 40 host cell proteins that interact with PEDV M protein using the proximity-labeling enzyme APEX2. Co-immunoprecipitation subsequently confirmed interactions between PEDV M protein and five host cell proteins, two of which (S100A11 and PPID) were involved in down-regulating virus replication in infected cells. This study is significant in that it formulates a strategy to provide new information about the mechanisms relating to the novel functions of PEDV M protein.

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