4.5 Article

Proteomic analysis of host cellular proteins co-immunoprecipitated with duck enteritis virus gC

期刊

JOURNAL OF PROTEOMICS
卷 245, 期 -, 页码 -

出版社

ELSEVIER
DOI: 10.1016/j.jprot.2021.104281

关键词

Duck enteritis virus; Proteomic probe; Host protein; coIP-MS; MS; Membrane-bound split-ubiquitin yeast two-hybrid system; Bimolecular fluorescence complementation

资金

  1. National Natural Science Foundation of China [31670150]
  2. National Key Research and Development Program of China [2016YFD0500107]

向作者/读者索取更多资源

This study confirmed the direct interaction between the gC protein of duck enteritis virus and seven host proteins (GNG2, AR1H1, PPP2CA, UBE2I, MCM5, NUBP1, HN1), further validated through MbYTH and BiFC analyses. These findings shed light on how DEV manipulates cellular machinery, enhancing our understanding of DEV pathogenesis.
Duck enteritis virus (DEV), the causative agent of duck viral enteritis, causes a contagious, lethal viral disease in Anseriformes (waterfowls). In virus infection, host-virus interaction plays a crucial role in virus replication and pathogenesis. In our previous study, mRFP was fused with the C-terminus of DEV glycoprotein C (gC) to construct a fluorescent-tag DEV virus rgCRFP. In the current study, fluorescent fusion protein (gC-mRFP) was used as the proteomic probe. Co-immunoprecipitation and mass spectrometric analysis of proteins from rgCRFPinfected chicken embryo fibroblasts using commercial anti-RFP antibody led to the identification of a total of 21 gC interacting host proteins. Out of these 21 proteins, the interaction of seven host proteins (GNG2, AR1H1, PPP2CA, UBE2I, MCM5, NUBP1, HN1) with DEV gC protein was validated using membrane-bound split-ubiquitin yeast two-hybrid system (MbYTH) and bimolecular fluorescence complementation (BiFC) analyses. It indicated direct interaction between these proteins with DEV gC protein. This study has furthered the current understanding of DEV virus infection and pathogenesis. Significance: gC is an crucial glycoprotein of duck enteritis virus that plays an important role in the viral life cycle. Uncovering the interaction between virus-host is very important to elucidate the pathogenic mechanism of the virus. In this study, host factors interacting with DEV gC have been discerned. And seven host proteins (GNG2, AR1H1, PPP2CA, UBE2I, MCM5, NUBP1, HN1) have been further validated to interact with DEV gC using MbYTH and BiFC analyses. These outcomes could shed light on how DEV manipulates the cellular machinery, which could further our understanding of DEV pathogenesis.

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