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

被引:16
作者
Dong, Shijuan [1 ,2 ]
Wang, Ruiyang [1 ]
Yu, Ruisong [1 ]
Chen, Bingqing [1 ,2 ]
Si, Fusheng [1 ,2 ]
Xie, Chunfang [1 ]
Li, Zhen [1 ,2 ,3 ]
机构
[1] Shanghai Acad Agr Sci SAAS, Inst Anim Sci & Vet Sci, Shanghai Key Lab Agr Genet & Breeding, Shanghai, Peoples R China
[2] Shanghai Acad Agr Sci SAAS, Shanghai Engn Res Ctr Breeding Pigs, Shanghai, Peoples R China
[3] Shanghai Acad Agr Sci, Inst Anim Husb & Vet Sci, Shanghai 201106, Peoples R China
关键词
PEDV; Membrane protein; Cellular proteins; APEX2; Interaction; EPIDEMIC-DIARRHEA-VIRUS; BINDING PROTEIN; CELLS; SUPPRESSION; SEQUENCE;
D O I
10.1016/j.jprot.2021.104191
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
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.
引用
收藏
页数:9
相关论文
共 30 条
  • [1] The V proteins of paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-β promoter
    Andrejeva, J
    Childs, KS
    Young, DF
    Carlos, TS
    Stock, N
    Goodbourn, S
    Randall, RE
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (49) : 17264 - 17269
  • [2] Porcine epidemic diarrhea virus inhibits dsRNA-induced interferon-β production in porcine intestinal epithelial cells by blockade of the RIG-I-mediated pathway
    Cao, Liyan
    Ge, Xuying
    Gao, Yu
    Herrler, Georg
    Ren, Yudong
    Ren, Xiaofeng
    Li, Guangxing
    [J]. VIROLOGY JOURNAL, 2015, 12
  • [3] Complete Genome Sequence of a Porcine Epidemic Diarrhea Virus Variant
    Chen, Jianfei
    Liu, Xiaozhen
    Shi, Da
    Shi, Hongyan
    Zhang, Xin
    Feng, Li
    [J]. JOURNAL OF VIROLOGY, 2012, 86 (06) : 3408 - 3408
  • [4] Cyclophilin 40 alters UVA-induced apoptosis and mitochondrial ROS generation in keratinocytes
    Jandova, Jana
    Janda, Jaroslav
    Sligh, James E.
    [J]. EXPERIMENTAL CELL RESEARCH, 2013, 319 (05) : 750 - 760
  • [5] Completion of the porcine epidemic diarrhoea coronavirus (PEDV) genome sequence
    Kocherhans, R
    Bridgen, A
    Ackermann, M
    Tobler, K
    [J]. VIRUS GENES, 2001, 23 (02) : 137 - 144
  • [6] Directed evolution of APEX2 for electron microscopy and proximity labeling
    Lam, Stephanie S.
    Martell, Jeffrey D.
    Kamer, Kimberli J.
    Deerinck, Thomas J.
    Ellisman, Mark H.
    Mootha, Vamsi K.
    Ting, Alice Y.
    [J]. NATURE METHODS, 2015, 12 (01) : 51 - 54
  • [7] Manipulation of the Porcine Epidemic Diarrhea Virus Genome Using Targeted RNA Recombination
    Li, Chunhua
    Li, Zhen
    Zou, Yong
    Wicht, Oliver
    van Kuppeveld, Frank J. M.
    Rottier, Peter J. M.
    Bosch, Berend Jan
    [J]. PLOS ONE, 2013, 8 (08):
  • [8] S100 Calcium Binding Protein A11 (S100A11) Promotes The Proliferation, Migration And Invasion Of Cervical Cancer Cells, And Activates Wnt/β-Catenin Signaling
    Meng, Man
    Sang, Lin
    Wang, Xiangyu
    [J]. ONCOTARGETS AND THERAPY, 2019, 12 : 8675 - 8685
  • [9] Proteomics of Primary Cilia by Proximity Labeling
    Mick, David U.
    Rodrigues, Rachel B.
    Leib, Ryan D.
    Adams, Christopher M.
    Chien, Allis S.
    Gygi, Steven P.
    Nachury, Maxence V.
    [J]. DEVELOPMENTAL CELL, 2015, 35 (04) : 497 - 512
  • [10] Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene
    Oka, Tomoichiro
    Saif, Linda J.
    Marthaler, Douglas
    Esseili, Malak A.
    Meulia, Tea
    Lin, Chun-Ming
    Vlasova, Anastasia N.
    Jung, Kwonil
    Zhang, Yan
    Wang, Qiuhong
    [J]. VETERINARY MICROBIOLOGY, 2014, 173 (3-4) : 258 - 269