title: Proteomics analysis of EV71-infected cells reveals the involvement of host protein NEDD4L in EV71 replication.
abstract: Enterovirus 71 (EV71) is a human enterovirus that has seriously affected the Asia-Pacific area for the past two decades. EV71 infection can result in mild hand-foot-and-mouth disease and herpangina and may occasionally lead to severe neurological complications in children. However, the specific biological processes that become altered during EV71 infection remain unclear. To further explore host responses upon EV71 infection, we identified proteins differentially expressed in EV71-infected human glioblastoma SF268 cells using isobaric mass tag (iTRAQ) labeling coupled with multidimensional liquid chromatography-mass spectrometry (LC-MS/MS). Network analysis of proteins altered in cells infected with EV71 revealed that the changed biological processes are related to protein and ion transport, regulation of protein degradation, and homeostatic processes. We confirmed that the levels of NEDD4L and PSMF1 were increased and reduced, respectively, in EV71-infected cells compared to mock-infected control cells. To determine the physiological relevance of our findings, we investigated the consequences of EV71 infection in cells with NEDD4L or PSMF1 depletion. We found that the depletion of NEDD4L significantly reduced the replication of EV71, whereas PSMF1 knockdown enhanced EV71 replication. Collectively, our findings provide the first evidence of proteome-wide dysregulation by EV71 infection and suggest a novel role for the host protein NEDD4L in the replication of this virus.
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Rei-Lin
authors.initials: RL
authors.lastname: Kuo
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Ya-Han
authors.initials: YH
authors.lastname: Lin
authors.affiliation: ‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|§Department of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Robert Yung-Liang
authors.initials: RY
authors.lastname: Wang
authors.affiliation: ∥Molecular Medicine Research Center, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Chia-Wei
authors.initials: CW
authors.lastname: Hsu
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Yi-Ting
authors.initials: YT
authors.lastname: Chiu
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Hsing-I
authors.initials: HI
authors.lastname: Huang
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Li-Ting
authors.initials: LT
authors.lastname: Kao
authors.affiliation: ∥Molecular Medicine Research Center, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Jau-Song
authors.initials: JS
authors.lastname: Yu
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|‡Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|⊥Clinical Virology Laboratory, Linkou Chang Gung Memorial Hospital, Tao-Yuan 333, Taiwan.
authors.forename: Shin-Ru
authors.initials: SR
authors.lastname: Shih
authors.affiliation: †Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|∥Molecular Medicine Research Center, Chang Gung University, Tao-Yuan 333, Taiwan.
authors.forename: Chih-Ching
authors.initials: CC
authors.lastname: Wu
mesh_terms: D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D019295:Computational Biology; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D029821:Enterovirus A, Human / Q000502:physiology*; D004769:Enterovirus Infections / Q000503:physiopathology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D055785:Gene Knockdown Techniques; D006801:Humans; D015151:Immunoblotting; D000075702:Nedd4 Ubiquitin Protein Ligases; D046988:Proteasome Endopeptidase Complex; D011506:Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053719:Tandem Mass Spectrometry; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology*
chemical_list: D017931:DNA Primers; D056827:Endosomal Sorting Complexes Required for Transport; C121623:PSMF1 protein, human; D011506:Proteins; D000075702:Nedd4 Ubiquitin Protein Ligases; C000618485:Nedd4L protein, human; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex
keywords: IFN-β; NEDD4L; PSMF1; enterovirus 71; iTRAQ