title: Proteomic analysis of mitochondria in respiratory epithelial cells infected with human respiratory syncytial virus and functional implications for virus and cell biology.
abstract: OBJECTIVE
  The aim of this study was to quantitatively characterise the mitochondrial proteome of airway epithelial cells infected with human respiratory syncytial virus (HRSV), a major cause of paediatric illness.
  
  
  METHODS
  Quantitative proteomics, underpinned by stable isotope labelling with amino acids in cell culture, coupled to LC-MS/MS, was applied to mitochondrial fractions prepared from HRSV-infected and mock-infected cells 12 and 24 h post-infection. Datasets were analysed using ingenuity pathway analysis, and the results were validated and characterised using bioimaging, targeted inhibition and gene depletion.
  
  
  RESULTS
  The data quantitatively indicated that antiviral signalling proteins converged on mitochondria during HRSV infection. The mitochondrial receptor protein Tom70 was found to act in an antiviral manner, while its chaperone, Hsp90, was confirmed to be a positive viral factor. Proteins associated with different organelles were also co-enriched in the mitochondrial fractions from HRSV-infected cells, suggesting that alterations in organelle dynamics and membrane associations occur during virus infection.
  
  
  CONCLUSIONS
  Protein and pathway-specific alterations occur to the mitochondrial proteome in a spatial and temporal manner during HRSV infection, suggesting that this organelle may have altered functions. These could be targeted as part of potential therapeutic strategies to disrupt virus biology.
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, UK.
authors.forename: Diane C
authors.initials: DC
authors.lastname: Munday
authors.forename: Gareth
authors.initials: G
authors.lastname: Howell
authors.forename: John N
authors.initials: JN
authors.lastname: Barr
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D002460:Cell Line; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000378:metabolism* / Q000821:virology; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics; D018357:Respiratory Syncytial Virus Infections / Q000378:metabolism* / Q000821:virology; D018113:Respiratory Syncytial Virus, Human; D012137:Respiratory System / Q000378:metabolism* / Q000821:virology; D053719:Tandem Mass Spectrometry
chemical_list: D018841:HSP90 Heat-Shock Proteins; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D020543:Proteome; C402118:TOMM70 protein, human
keywords: inhibition; mitochondria; proteomics; respiratory syncytial virus; siRNA