title: Proteomic analysis at the subcellular level for host targets against influenza A virus (H1N1).
abstract: Influenza viruses (IVs) trigger a series of intracellular signaling events and induce complex cellular responses from the infected host cell. Accumulating evidence suggests that host cell proteins play an essential role in viral propagation and represent novel antiviral therapeutic targets. Subcellular proteomic technology provides a method for understanding regional differences at the protein level. The present study, which utilized subcellular proteomic technology, aimed to identify host cell proteins involved in influenza virus (HIN1) infection. Two-dimensional gel electrophoresis (2-DE) combined with mass spectrum (MS) was performed on protein extracts from the nuclei, cytoplasm, and mitochondria of infected and control human lung epithelial cells (A549). In total, 112 differentially expressed protein molecules were identified; 80 protein spots were successfully validated using MS. The differential expression of ISG15, MIF, PDCD5, and UCHL1 was confirmed by western blot. Furthermore, antisense oligodeoxyribonucleotide (ODN) targeting ISG15, MIF, PDCD5, and UCHL1 significantly mitigated HIN1 propagation, cytopathic effects, vRNA by RT-qPCR, and rescued cell viability in A549 cells. Taken together, the differentially expressed proteins identified in this study might provide novel targets for anti-influenza drug development.
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, PR China.
authors.forename: Haibao
authors.initials: H
authors.lastname: Zhao
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Kang
authors.initials: K
authors.lastname: Li
authors.forename: Xiaoran
authors.initials: X
authors.lastname: Ding
authors.forename: Ruxian
authors.initials: R
authors.lastname: Lin
authors.forename: Yongjie
authors.initials: Y
authors.lastname: Ma
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.forename: Zhiyin
authors.initials: Z
authors.lastname: Zhong
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qian
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Bo
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhou
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000737:chemistry; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000821:virology; D015967:Gene Expression Regulation, Viral; D020868:Gene Silencing; D054884:Host-Pathogen Interactions; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000502:physiology*; D008168:Lung; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000502:physiology; D040901:Proteomics; D012367:RNA, Viral / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000737:chemistry; D014779:Virus Replication
chemical_list: D017077:Culture Media, Conditioned; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D011506:Proteins; D012367:RNA, Viral
keywords: H1N1; Host target; Influenza virus; Proteomic analysis