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Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1

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...

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Autores principales: Guo, Hui-Chen, Jin, Ye, Han, Shi-Chong, Sun, Shi-Qi, Wei, Yan-Quan, Liu, Xian-Ji, Feng, Xia, Liu, Ding Xiang, Liu, Xiang-Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498813/
https://www.ncbi.nlm.nih.gov/pubmed/26161868
http://dx.doi.org/10.1371/journal.pone.0132384
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author Guo, Hui-Chen
Jin, Ye
Han, Shi-Chong
Sun, Shi-Qi
Wei, Yan-Quan
Liu, Xian-Ji
Feng, Xia
Liu, Ding Xiang
Liu, Xiang-Tao
author_facet Guo, Hui-Chen
Jin, Ye
Han, Shi-Chong
Sun, Shi-Qi
Wei, Yan-Quan
Liu, Xian-Ji
Feng, Xia
Liu, Ding Xiang
Liu, Xiang-Tao
author_sort Guo, Hui-Chen
collection PubMed
description 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.
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spelling pubmed-44988132015-07-17 Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1 Guo, Hui-Chen Jin, Ye Han, Shi-Chong Sun, Shi-Qi Wei, Yan-Quan Liu, Xian-Ji Feng, Xia Liu, Ding Xiang Liu, Xiang-Tao PLoS One Research Article 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. Public Library of Science 2015-07-10 /pmc/articles/PMC4498813/ /pubmed/26161868 http://dx.doi.org/10.1371/journal.pone.0132384 Text en © 2015 Guo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Guo, Hui-Chen
Jin, Ye
Han, Shi-Chong
Sun, Shi-Qi
Wei, Yan-Quan
Liu, Xian-Ji
Feng, Xia
Liu, Ding Xiang
Liu, Xiang-Tao
Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title_full Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title_fullStr Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title_full_unstemmed Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title_short Quantitative Proteomic Analysis of BHK-21 Cells Infected with Foot-and-Mouth Disease Virus Serotype Asia 1
title_sort quantitative proteomic analysis of bhk-21 cells infected with foot-and-mouth disease virus serotype asia 1
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498813/
https://www.ncbi.nlm.nih.gov/pubmed/26161868
http://dx.doi.org/10.1371/journal.pone.0132384
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