title: Comparative proteomic analyses demonstrate enhanced interferon and STAT-1 activation in reovirus T3D-infected HeLa cells.
abstract: As obligate intracellular parasites, viruses are exclusively and intimately dependent upon their host cells for replication. During replication viruses induce profound changes within cells, including: induction of signaling pathways, morphological changes, and cell death. Many such cellular perturbations have been analyzed at the transcriptomic level by gene arrays and recent efforts have begun to analyze cellular proteomic responses. We recently described comparative stable isotopic (SILAC) analyses of reovirus, strain type 3 Dearing (T3D)-infected HeLa cells. For the present study we employed the complementary labeling strategy of iTRAQ (isobaric tags for relative and absolute quantitation) to examine HeLa cell changes induced by T3D, another reovirus strain, type 1 Lang, and UV-inactivated T3D (UV-T3D). Triplicate replicates of cytosolic and nuclear fractions identified a total of 2375 proteins, of which 50, 57, and 46 were significantly up-regulated, and 37, 26, and 44 were significantly down-regulated by T1L, T3D, and UV-T3D, respectively. Several pathways, most notably the Interferon signaling pathway and the EIF2 and ILK signaling pathways, were induced by virus infection. Western blots confirmed that cells were more strongly activated by live T3D as demonstrated by elevated levels of key proteins like STAT-1, ISG-15, IFIT-1, IFIT-3, and Mx1. This study expands our understanding of reovirus-induced host responses.
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, University of Manitoba Winnipeg, MB, Canada.
authors.forename: Peyman
authors.initials: P
authors.lastname: Ezzati
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Manitoba Winnipeg, MB, Canada.
authors.forename: Krysten
authors.initials: K
authors.lastname: Komher
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Manitoba Winnipeg, MB, Canada.
authors.forename: Giulia
authors.initials: G
authors.lastname: Severini
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, University of Manitoba Winnipeg, MB, Canada ; Department of Medical Microbiology, Faculty of Medicine, University of Manitoba Winnipeg, MB, Canada ; Manitoba Institute of Child Health, John Buhler Research Centre Winnipeg, MB, Canada.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Coombs
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D012089:Mammalian orthoreovirus 3 / Q000235:genetics / Q000502:physiology*; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D016601:RNA-Binding Proteins; D012088:Reoviridae Infections / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D016207:Cytokines; C484290:IFIT1 protein, human; C116468:IFIT3 protein, human; D047908:Intracellular Signaling Peptides and Proteins; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D016601:RNA-Binding Proteins; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D014452:Ubiquitins; C074034:ISG15 protein, human; D007372:Interferons
keywords: RNA virus; bioinformatics; cell signaling; host cell alterations; liquid chromatography; mass spectrometry; virus infection