High-throughput quantitative proteomic analysis of dengue virus type 2 infected A549 cells

Han-Chen Chiu, Holger Hannemann, Kate J. Heesom, David A. Matthews, Andrew D. Davidson*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

32 Citations (Scopus)

Abstract

Disease caused by dengue virus is a global health concern with up to 390 million individuals infected annually worldwide. There are no vaccines or antiviral compounds available to either prevent or treat dengue disease which may be fatal. To increase our understanding of the interaction of dengue virus with the host cell, we analyzed changes in the proteome of human A549 cells in response to dengue virus type 2 infection using stable isotope labelling in cell culture (SILAC) in combination with high-throughput mass spectrometry (MS). Mock and infected A549 cells were fractionated into nuclear and cytoplasmic extracts before analysis to identify proteins that redistribute between cellular compartments during infection and reduce the complexity of the analysis. We identified and quantified 3098 and 2115 proteins in the cytoplasmic and nuclear fractions respectively. Proteins that showed a significant alteration in amount during infection were examined using gene enrichment, pathway and network analysis tools. The analyses revealed that dengue virus infection modulated the amounts of proteins involved in the interferon and unfolded protein responses, lipid metabolism and the cell cycle. The SILAC-MS results were validated for a select number of proteins over a time course of infection by Western blotting and immunofluorescence microscopy. Our study demonstrates for the first time the power of SILAC-MS for identifying and quantifying novel changes in cellular protein amounts in response to dengue virus infection.

Original languageEnglish
Article number93305
Pages (from-to)e93305
Number of pages22
JournalPLoS ONE
Volume9
Issue number3
DOIs
Publication statusPublished - 26 Mar 2014

Keywords

  • GENE TRANSCRIPT ABUNDANCE
  • UNFOLDED PROTEIN RESPONSE
  • HUMAN ENDOTHELIAL-CELLS
  • WEST-NILE-VIRUS
  • NF-KAPPA-B
  • NUCLEAR-LOCALIZATION
  • HEMORRHAGIC-FEVER
  • EXOCYTOTIC CARRIERS
  • PERIPHERAL-BLOOD
  • DENDRITIC CELLS

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