title: Identification of host proteins involved in Japanese encephalitis virus infection by quantitative proteomics analysis.
abstract: Japanese encephalitis virus (JEV) enters host cells via receptor-mediated endocytosis and replicates in the cytoplasm of infected cells. To study virus-host cell interactions, we performed a SILAC-based quantitative proteomics study of JEV-infected HeLa cells using a subcellular fractionation strategy. We identified 158 host proteins as differentially regulated by JEV (defined as exhibiting a greater than 1.5-fold change in protein abundance upon JEV infection). The mass spectrometry quantitation data for selected proteins were validated by Western blot and immunofluorescence confocal microscopy. Bioinformatics analyses were used to generate JEV-regulated host response networks consisting of regulated proteins, which included 35 proteins that were newly added based on the results of this study. The JEV infection-induced host response was found to be coordinated primarily through the immune response process, the ubiquitin-proteasome system (UPS), the intracellular membrane system, and lipid metabolism-related proteins. Protein functional studies of selected host proteins using RNA interference-based techniques were carried out in HeLa cells infected with an attenuated or a highly virulent strain of JEV. We demonstrated that the knockdown of interferon-induced transmembrane protein 3 (IFITM3), Ran-binding protein 2 (RANBP2), sterile alpha motif domain-containing protein 9 (SAMD9) and vesicle-associated membrane protein 8 (VAMP8) significantly increased JEV replication. The results presented here not only promote a better understanding of the host response to JEV infection but also highlight multiple potential targets for the development of antiviral agents.
authors: State Key Laboratory of Virology, Wuhan University, Wuhan, China.
authors: Lei-Ke
authors: LK
authors: Zhang
authors: Fan
authors: F
authors: Chai
authors: Hao-Yu
authors: HY
authors: Li
authors: Gengfu
authors: G
authors: Xiao
authors: Lin
authors: L
authors: Guo
mesh_terms: D000943:Antigens, Differentiation / Q000235:genetics* / Q000276:immunology; D002458:Cell Fractionation; D004664:Encephalitis Virus, Japanese / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones / Q000235:genetics* / Q000276:immunology; D028861:Nuclear Pore Complex Proteins / Q000235:genetics* / Q000276:immunology; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000235:genetics* / Q000276:immunology; D040901:Proteomics; D050683:R-SNARE Proteins / Q000235:genetics* / Q000276:immunology; D015398:Signal Transduction; D014779:Virus Replication
chemical_list: D000943:Antigens, Differentiation; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D028861:Nuclear Pore Complex Proteins; D011506:Proteins; D050683:R-SNARE Proteins; C514443:SAMD9 protein, human; C112938:VAMP8 protein, human; C062010:leu-13 antigen; C094493:ran-binding protein 2; D046988:Proteasome Endopeptidase Complex