Quantitative Proteomics Using Stable Isotope Labeling with Amino Acids in Cell Culture Reveals Protein and Pathway Regulation in Porcine Circovirus Type 2 Infected PK-15 Cells

The infection of host cells by porcine circovirus type 2 (PCV2) leads to extensive modulation of the gene expression levels of target cells. To uncover the pathogenesis and virus-host interactions of PCV2, a quantitative proteomic study using the stable isotope labeling with amino acids in cell cult...

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Bibliographic Details
Published in:Journal of proteome research 2012-02, Vol.11 (2), p.995-1008
Main Authors: Fan, Huiying, Ye, Yu, Luo, Yongwen, Tong, Tiezhu, Yan, Guangrong, Liao, Ming
Format: Article
Language:English
Subjects:
Animals
Cell Line
Circoviridae Infections - metabolism
Circoviridae Infections - microbiology
Circovirus - metabolism
Down-Regulation
Host-Pathogen Interactions
Isotope Labeling - methods
Protein Interaction Maps
Proteins - analysis
Proteins - chemistry
Proteins - classification
Proteome - analysis
Proteome - chemistry
Proteomics - methods
Signal Transduction
Swine
Up-Regulation
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Summary:The infection of host cells by porcine circovirus type 2 (PCV2) leads to extensive modulation of the gene expression levels of target cells. To uncover the pathogenesis and virus-host interactions of PCV2, a quantitative proteomic study using the stable isotope labeling with amino acids in cell culture (SILAC), coupled with mass spectrometry, was performed on PCV2-infected PK-15 cells. The SILAC-based approach identified 1341 proteins, 163 of which showed significant change in level at 72 h after infection (79 up-regulated and 84 down-regulated). The modulated proteins included a number of proteins involved in substrate transport, cytoskeletal changes, and the stress response. Changes in the expression levels of selected proteins were verified by Western blot analysis. Ingenuity Pathway Analysis was used to reveal protein and interactive pathway regulation in response to PCV2 infection. Functional network and pathway analyses could provide insights into the complexity and dynamics of virus–host cell interactions and may accelerate our understanding of the mechanisms of PCV2 infection.
ISSN:1535-3893
1535-3907
DOI:10.1021/pr200755d