title: Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1.
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.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Hui-Chen
authors.initials: HC
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Shi-Chong
authors.initials: SC
authors.lastname: Han
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Shi-Qi
authors.initials: SQ
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Yan-Quan
authors.initials: YQ
authors.lastname: Wei
authors.affiliation: Guangzhou Fitgene Biotechnology Co. Ltd. Guangzhou International Business Incubator A110, Guangzhou, Guangdong, China.
authors.forename: Xian-Ji
authors.initials: XJ
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China; School of Biological Sciences, Nanyang Technological University, Singapore.
authors.forename: Ding Xiang
authors.initials: DX
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology and OIE/National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Xiang-Tao
authors.initials: XT
authors.lastname: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002853:Chromatography, Liquid; D019295:Computational Biology; D015536:Down-Regulation; D005536:Foot-and-Mouth Disease / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000502:physiology*; D055785:Gene Knockdown Techniques; D005814:Genes, Viral; D015151:Immunoblotting; D007553:Isotope Labeling; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D015203:Reproducibility of Results; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D020543:Proteome; D034741:RNA, Small Interfering; D014764:Viral Proteins