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Quantitative Proteomics Analysis of the Hepatitis C Virus Replicon High-Permissive and Low-Permissive Cell Lines.

Ye F, Xin Z, Han W, Fan J, Yin B, Wu S, Yang W, Yuan J, Qiang B, Sun W, Peng X - PLoS ONE (2015)

Bottom Line: The development of the subgenome replicon model system significantly enhanced study of HCV.And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels.Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation.

View Article: PubMed Central - PubMed

Affiliation: The State Key Laboratory of Medical Molecular Biology, Department of Molecular Biology and Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Chronic hepatitis C virus (HCV) infection is one of the leading causes of severe hepatitis. The molecular mechanisms underlying HCV replication and pathogenesis remain unclear. The development of the subgenome replicon model system significantly enhanced study of HCV. However, the permissiveness of the HCV subgenome replicon greatly differs among different hepatoma cell lines. Proteomic analysis of different permissive cell lines might provide new clues in understanding HCV replication. In this study, to detect potential candidates that might account for the differences in HCV replication. Label-free and iTRAQ labeling were used to analyze the differentially expressed protein profiles between Huh7.5.1 wt and HepG2 cells. A total of 4919 proteins were quantified in which 114 proteins were commonly identified as differentially expressed by both quantitative methods. A total of 37 differential proteins were validated by qRT-PCR. The differential expression of Glutathione S-transferase P (GSTP1), Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), carboxylesterase 1 (CES1), vimentin, Proteasome activator complex subunit1 (PSME1), and Cathepsin B (CTSB) were verified by western blot. And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels. Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation. These results highlight the potential role of CTSB and vimentin in virus replication.

No MeSH data available.


Related in: MedlinePlus

Classification of differentially expressed proteins based on GO annotation.(A) Categorization based upon biological process analysis of total differential protein expression. (B) Categorization based upon molecular function analysis of total differential protein expression. The distribution frequencies in regard to the specified categories within the given charts are indicated in % of the total number of protein entries.
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pone.0142082.g003: Classification of differentially expressed proteins based on GO annotation.(A) Categorization based upon biological process analysis of total differential protein expression. (B) Categorization based upon molecular function analysis of total differential protein expression. The distribution frequencies in regard to the specified categories within the given charts are indicated in % of the total number of protein entries.

Mentions: To gain a functional understanding of the possible roles of the differentially expressed proteins, GO annotation was performed using “biological process” and “molecular function”. The 674 differential proteins were clustered according to their biological processes and molecular function (Fig 3). Proteins were involved in several biological processes that are relevant for HCV infection, especially growth and inflammation. A significant number of identified proteins (29.8%) were engaged in metabolic process, while the next top five biological process categories were cellular process (14.2%), cell communication (9.3%), transport (7.5%), immune system process (6.4%) and developmental process (6.2%) (Fig 3A). In the molecular function distribution category, the majority of the proteins were associated with three functions: catalytic activity (40.1%), binding (29.2%) and structural molecule activity (10.4%) (Fig 3B).


Quantitative Proteomics Analysis of the Hepatitis C Virus Replicon High-Permissive and Low-Permissive Cell Lines.

Ye F, Xin Z, Han W, Fan J, Yin B, Wu S, Yang W, Yuan J, Qiang B, Sun W, Peng X - PLoS ONE (2015)

Classification of differentially expressed proteins based on GO annotation.(A) Categorization based upon biological process analysis of total differential protein expression. (B) Categorization based upon molecular function analysis of total differential protein expression. The distribution frequencies in regard to the specified categories within the given charts are indicated in % of the total number of protein entries.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4636247&req=5

pone.0142082.g003: Classification of differentially expressed proteins based on GO annotation.(A) Categorization based upon biological process analysis of total differential protein expression. (B) Categorization based upon molecular function analysis of total differential protein expression. The distribution frequencies in regard to the specified categories within the given charts are indicated in % of the total number of protein entries.
Mentions: To gain a functional understanding of the possible roles of the differentially expressed proteins, GO annotation was performed using “biological process” and “molecular function”. The 674 differential proteins were clustered according to their biological processes and molecular function (Fig 3). Proteins were involved in several biological processes that are relevant for HCV infection, especially growth and inflammation. A significant number of identified proteins (29.8%) were engaged in metabolic process, while the next top five biological process categories were cellular process (14.2%), cell communication (9.3%), transport (7.5%), immune system process (6.4%) and developmental process (6.2%) (Fig 3A). In the molecular function distribution category, the majority of the proteins were associated with three functions: catalytic activity (40.1%), binding (29.2%) and structural molecule activity (10.4%) (Fig 3B).

Bottom Line: The development of the subgenome replicon model system significantly enhanced study of HCV.And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels.Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation.

View Article: PubMed Central - PubMed

Affiliation: The State Key Laboratory of Medical Molecular Biology, Department of Molecular Biology and Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Chronic hepatitis C virus (HCV) infection is one of the leading causes of severe hepatitis. The molecular mechanisms underlying HCV replication and pathogenesis remain unclear. The development of the subgenome replicon model system significantly enhanced study of HCV. However, the permissiveness of the HCV subgenome replicon greatly differs among different hepatoma cell lines. Proteomic analysis of different permissive cell lines might provide new clues in understanding HCV replication. In this study, to detect potential candidates that might account for the differences in HCV replication. Label-free and iTRAQ labeling were used to analyze the differentially expressed protein profiles between Huh7.5.1 wt and HepG2 cells. A total of 4919 proteins were quantified in which 114 proteins were commonly identified as differentially expressed by both quantitative methods. A total of 37 differential proteins were validated by qRT-PCR. The differential expression of Glutathione S-transferase P (GSTP1), Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), carboxylesterase 1 (CES1), vimentin, Proteasome activator complex subunit1 (PSME1), and Cathepsin B (CTSB) were verified by western blot. And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels. Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation. These results highlight the potential role of CTSB and vimentin in virus replication.

No MeSH data available.


Related in: MedlinePlus