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Abstract
Stable isotope labeling with amino acids in cell culture (SILAC) was used to quantitatively study the host cell gene expression profile, in order to achieve an unbiased overview of the protein expression changes in BHK-21 cells infected with FMDV serotype Asia 1. The SILAC-based approach identified overall 2,141 proteins, 153 of which showed significant alteration in the expression level 6 h post FMDV infection (57 up-regulated and 96 down-regulated). Among these proteins, six cellular proteins, including three down-regulated (VPS28, PKR, EVI5) and three up-regulated (LYPLA1, SEC62 and DARs), were selected according to the significance of the changes and/or the relationship with PKR. The expression level and pattern of the selected proteins were validated by immunoblotting and confocal microscopy. Furthermore, the functions of these cellular proteins were assessed by small interfering RNA-mediated depletion, and their functional importance in the replication of FMDV was demonstrated by western blot, reverse transcript PCR (RT-PCR) and 50% Tissue Culture Infective Dose (TCID50). The results suggest that FMDV infection may have effects on the expression of specific cellular proteins to create more favorable conditions for FMDV infection. This study provides novel data that can be utilized to understand the interactions between FMDV and the host cell.
Highlights
Foot-and-mouth disease (FMD) is one of the most economically important diseases of clovenhoofed animals because it severely compromises livestock production, resulting in high economic losses and international restrictions on the export of animals and animal products [1]
The results showed that the abundance and expression patterns of 153 proteins were significantly changed upon foot-and-mouth disease virus (FMDV) infection
The FMDV protein expression reached the peak at 8h p.i., the expression of β-actin as a control was markedly decreased at this time point, suggesting virus-induced suppression of host cell protein synthesis and lysis or death of a majority of the cells at 8h p.i
Summary
Foot-and-mouth disease (FMD) is one of the most economically important diseases of clovenhoofed animals because it severely compromises livestock production, resulting in high economic losses and international restrictions on the export of animals and animal products [1]. The causative agent is the foot-and-mouth disease virus (FMDV) belonging to genus Aphthovirus of the family Picornaviridae, which is a non-enveloped virus possessing positive polarity. Proteomic Analysis of FMDV-Infected BHK-21 Cells the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author are articulated in the ‘author contributions’ section
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