Limits...
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

The overall workflow.Huh7.5.1 wt and HepG2 whole-cell proteins were harvested and analyzed using the Label-free and iTRAQ proteomic approaches, and the differential proteins were verified by qRT-PCR and western blot. The functions of selected differential proteins in HCV replication were analyzed further.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4636247&req=5

pone.0142082.g001: The overall workflow.Huh7.5.1 wt and HepG2 whole-cell proteins were harvested and analyzed using the Label-free and iTRAQ proteomic approaches, and the differential proteins were verified by qRT-PCR and western blot. The functions of selected differential proteins in HCV replication were analyzed further.

Mentions: The inter-sample analysis is visualized using a volcano plot comparing the fold change and t-test p-value (Fig 1). A total of 393 proteins were determined to be modulated more than 2-fold with t-test p-value less than 0.05 (S1 Table).


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)

The overall workflow.Huh7.5.1 wt and HepG2 whole-cell proteins were harvested and analyzed using the Label-free and iTRAQ proteomic approaches, and the differential proteins were verified by qRT-PCR and western blot. The functions of selected differential proteins in HCV replication were analyzed further.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0142082.g001: The overall workflow.Huh7.5.1 wt and HepG2 whole-cell proteins were harvested and analyzed using the Label-free and iTRAQ proteomic approaches, and the differential proteins were verified by qRT-PCR and western blot. The functions of selected differential proteins in HCV replication were analyzed further.
Mentions: The inter-sample analysis is visualized using a volcano plot comparing the fold change and t-test p-value (Fig 1). A total of 393 proteins were determined to be modulated more than 2-fold with t-test p-value less than 0.05 (S1 Table).

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