title: Subcellular proteomic analysis of human host cells infected with H3N2 swine influenza virus.
abstract: Cross-species transmissions of swine influenza viruses (SIVs) raise great public health concerns. In this study, subcellular proteomic profiles of human A549 cells inoculated with H3N2 subtype SIV were used to characterize dynamic cellular responses to infection. By 2DE and MS, 27 differentially expressed (13 upregulated, 14 downregulated) cytoplasmic proteins and 20 differentially expressed (13 upregulated, 7 downregulated) nuclear proteins were identified. Gene ontology analysis suggested that these differentially expressed proteins were mainly involved in cell death, stress response, lipid metabolism, cell signaling, and RNA PTMs. Moreover, 25 corresponding genes of the differentially expressed proteins were quantitated by real time RT-PCR to examine the transcriptional profiles between mock- and virus-infected A549 cells. Western blot analysis confirmed that changes in abundance of identified cellular proteins heterogeneous nuclear ribonucleoprotein (hnRNP) U, hnRNP C, ALDH1A1, tryptophanyl-tRNA synthetase, IFI35, and HSPB1 in H3N2 SIV-infected cells were consistent with results of 2DE analysis. By confocal microscopy, nucleus-to-cytoplasm translocation of hnRNP C and colocalization between the viral nonstructural protein 1 and hnRNP C as well as N-myc (and STAT) interactor were observed upon infection. Ingenuity Pathway Analysis revealed that cellular proteins altered during infection were grouped mainly into NFκB and interferon signaling networks. Collectively, these identified subcellular constituents provide an important framework for understanding host/SIV interactions and underlying mechanisms of SIV cross-species infection and pathogenesis.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, P. R. China; State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University, Hangzhou, P. R. China.
authors.forename: Xiaopeng
authors.initials: X
authors.lastname: Wu
authors.forename: Sanying
authors.initials: S
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Jinyang
authors.initials: J
authors.lastname: Zhang
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Sun
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000502:physiology*; D042541:Intracellular Space / Q000737:chemistry* / Q000378:metabolism / Q000821:virology*; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000145:classification / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D059467:Transcriptome
chemical_list: D011506:Proteins; D020543:Proteome
keywords: 2DE; A549 cells; Interaction; Microbiology; Subcellular proteomics; Swine influenza