tetano
Editor, Senior Moderator
Proteomics. 2013 Sep 30. doi: 10.1002/pmic.201300180. [Epub ahead of print]
Subcellular Proteomic Analysis of Human Host Cells Infected with H3N2 Swine Influenza Virus.
Wu X, Wang S, Yu Y, Zhang J, Sun Z, Yan Y, Zhou J.
Source
Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, 310058, PR China; State Key Laboratory.
Abstract
Cross-species transmissions of swine influenza viruses (SIV) raise great public health concerns. Here, subcellular proteomic profiles of human A549 cells inoculated with H3N2 subtype SIV were used to characterize dynamic cellular responses to infection. By two-dimensional gel electrophoresis (2-DE) and mass spectrometry, 27 differentially expressed (13 up-regulated, 14 down-regulated) cytoplasmic proteins and 20 differentially expressed (13 up-regulated, 7 down-regulated) 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 post-transcriptional modifications. 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 hnRNP U, hnRNP C, ALDH1A1, WARS, IFI35 and HSPB1 in H3N2 SIV-infected cells were consistent with results of 2-DE analysis. By confocal microscopy, nucleus-to-cytoplasm translocation of hnRNP C and colocalization between the viral protein NS1 and hnRNP C as well as NMI 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. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
A549 cells, interaction, subcellular proteomics, swine influenza, two-dimensional gel electrophoresis
PMID:
24115376
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24115376
Subcellular Proteomic Analysis of Human Host Cells Infected with H3N2 Swine Influenza Virus.
Wu X, Wang S, Yu Y, Zhang J, Sun Z, Yan Y, Zhou J.
Source
Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, 310058, PR China; State Key Laboratory.
Abstract
Cross-species transmissions of swine influenza viruses (SIV) raise great public health concerns. Here, subcellular proteomic profiles of human A549 cells inoculated with H3N2 subtype SIV were used to characterize dynamic cellular responses to infection. By two-dimensional gel electrophoresis (2-DE) and mass spectrometry, 27 differentially expressed (13 up-regulated, 14 down-regulated) cytoplasmic proteins and 20 differentially expressed (13 up-regulated, 7 down-regulated) 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 post-transcriptional modifications. 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 hnRNP U, hnRNP C, ALDH1A1, WARS, IFI35 and HSPB1 in H3N2 SIV-infected cells were consistent with results of 2-DE analysis. By confocal microscopy, nucleus-to-cytoplasm translocation of hnRNP C and colocalization between the viral protein NS1 and hnRNP C as well as NMI 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. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
A549 cells, interaction, subcellular proteomics, swine influenza, two-dimensional gel electrophoresis
PMID:
24115376
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24115376