Originally published In Press as doi:10.1074/mcp.M700396-MCP200 on December 4, 2007.
Molecular & Cellular Proteomics 7:612-625, 2008.
© 2008 by The American Society for Biochemistry and Molecular Biology, Inc.
Research
Proteomics Analysis of Host Cells Infected with Infectious Bursal Disease Virus*,S
Xiaojuan Zheng ,
Lianlian Hong ,
Lixue Shi ,
Junqing Guo ,
Zhen Sun and
Jiyong Zhou , ,¶,||
From the Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, Hangzhou 310003, China, and ¶ Key Laboratory of conservation genetics and reproductive biology for endangered wild animals of the Ministry of Education, Zhejiang University, Hangzhou 310058, China
The effect of infectious bursal disease virus (IBDV) infection on cellular protein expression is essential for viral pathogenesis. To characterize the cellular response to IBDV infection, the differential proteomes of chicken embryo fibroblasts, with and without IBDV infection, were analyzed at different time points with two-dimensional gel electrophoresis (2-DE) followed by MALDI-TOF/TOF identification. Comparative analysis of multiple 2-DE gels revealed that the majority of protein expression changes appeared at 48 and 96 h after IBDV infection. Mass spectrometry identified 51 altered cellular proteins, including 13 up-regulated proteins and 38 down-regulated proteins 12–96 h after infection. Notably 2-DE analysis revealed that IBDV infection induced the increased expression of polyubiquitin, apolipoprotein A-I, heat shock 27-kDa protein 1, actins, tubulins, eukaryotic translation initiation factor 4A isoform 2, acidic ribosomal phosphoprotein, and ribosomal protein SA isoform 2. In addition, IBDV infection considerably suppressed those cellular proteins involved in ubiquitin-mediated protein degradation, energy metabolism, intermediate filaments, host translational apparatus, and signal transduction. Moreover 38 corresponding genes of the differentially expressed proteins were quantitated by real time RT-PCR to examine the transcriptional profiles between infected and uninfected chicken embryo fibroblasts. Western blot further confirmed the inhibition of Rho protein GDP dissociation inhibitor expression and the induction of polyubiquitin during IBDV infection. Subcellular distribution analysis of the cytoskeletal proteins vimentin and β-tubulin clearly demonstrated that IBDV infection induced the disruption of the vimentin network and microtubules late in IBDV infection. Thus, this work effectively provides useful dynamic protein-related information to facilitate further investigation of the underlying mechanism of IBDV infection and pathogenesis.
|| To whom correspondence should be addressed: Inst. of Preventive Veterinary Medicine, Zhejiang University, 268 Kaixuan Rd., Hangzhou 310029, China. Tel.: 86-571-8697-1698; Fax: 86-571-8697-1821; E-mail: jyzhou{at}zju.edu.cn

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Copyright © 2008 by the American Society for Biochemistry and Molecular Biology.
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