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Title: Proteome Analysis of Liver Cells Expressing a Full- Length Hepatitis C Virus (HCV) Replicon and Biopsy Specimens of Posttransplantation Liver from HCV-Infected Patients

Abstract

The development of a reproducible model system for the study of Hepatitis C virus (HCV) infection has the potential to significantly enhance the study of virus-host interactions and provide future direction for modeling the pathogenesis of HCV. While there are studies describing global gene expression changes associated with HCV infection, changes in the proteome have not been characterized. We report the first large scale proteome analysis of the highly permissive Huh-7.5 cell line containing a full length HCV replicon. We detected > 4,400 proteins in this cell line, including HCV replicon proteins, using multidimensional liquid chromatographic (LC) separations coupled to mass spectrometry (MS). The set of Huh-7.5 proteins confidently identified is, to our knowledge, the most comprehensive yet reported for a human cell line. Consistent with the literature, a comparison of Huh-7.5 cells (+) and (-) the HCV replicon identified expression changes of proteins involved in lipid metabolism. We extended these analyses to liver biopsy material from HCV-infected patients where > 1,500 proteins were detected from 2 {micro}g protein lysate using the Huh-7.5 protein database and the accurate mass and time (AMT) tag strategy. These findings demonstrate the utility of multidimensional proteome analysis of the HCV replicon model system formore » assisting the determination of proteins/pathways affected by HCV infection. Our ability to extend these analyses to the highly complex proteome of small liver biopsies with limiting protein yields offers the unique opportunity to begin evaluating the clinical significance of protein expression changes associated with HCV infection.« less

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
15016517
Report Number(s):
PNWD-SA-6678
Journal ID: ISSN 0022-538X; JOVIAM; 4393; TRN: US200513%%615
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of Virology, 79(12):7558-7569
Additional Journal Information:
Journal Volume: 79; Journal Issue: 12; Journal ID: ISSN 0022-538X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ANIMAL CELLS; BIOPSY; GENES; HEPATITIS; LIPIDS; LIVER; LIVER CELLS; MASS SPECTROSCOPY; METABOLISM; PATHOGENESIS; PATIENTS; PROTEINS; REPLICONS; SIMULATION; Environmental Molecular Sciences Laboratory

Citation Formats

Jacobs, Jon M., Diamond, Deborah L., Chan, Eric Y., Gritsenko, Marina A., Qian, Weijun, Stastna, Miroslava, Baas, Tracey, Camp, David G., Carithers, Jr., Robert L., Smith, Richard D., and Katze, Michael G. Proteome Analysis of Liver Cells Expressing a Full- Length Hepatitis C Virus (HCV) Replicon and Biopsy Specimens of Posttransplantation Liver from HCV-Infected Patients. United States: N. p., 2005. Web. doi:10.1128/JVI.79.12.7558-7569.2005.
Jacobs, Jon M., Diamond, Deborah L., Chan, Eric Y., Gritsenko, Marina A., Qian, Weijun, Stastna, Miroslava, Baas, Tracey, Camp, David G., Carithers, Jr., Robert L., Smith, Richard D., & Katze, Michael G. Proteome Analysis of Liver Cells Expressing a Full- Length Hepatitis C Virus (HCV) Replicon and Biopsy Specimens of Posttransplantation Liver from HCV-Infected Patients. United States. doi:10.1128/JVI.79.12.7558-7569.2005.
Jacobs, Jon M., Diamond, Deborah L., Chan, Eric Y., Gritsenko, Marina A., Qian, Weijun, Stastna, Miroslava, Baas, Tracey, Camp, David G., Carithers, Jr., Robert L., Smith, Richard D., and Katze, Michael G. Wed . "Proteome Analysis of Liver Cells Expressing a Full- Length Hepatitis C Virus (HCV) Replicon and Biopsy Specimens of Posttransplantation Liver from HCV-Infected Patients". United States. doi:10.1128/JVI.79.12.7558-7569.2005.
@article{osti_15016517,
title = {Proteome Analysis of Liver Cells Expressing a Full- Length Hepatitis C Virus (HCV) Replicon and Biopsy Specimens of Posttransplantation Liver from HCV-Infected Patients},
author = {Jacobs, Jon M. and Diamond, Deborah L. and Chan, Eric Y. and Gritsenko, Marina A. and Qian, Weijun and Stastna, Miroslava and Baas, Tracey and Camp, David G. and Carithers, Jr., Robert L. and Smith, Richard D. and Katze, Michael G.},
abstractNote = {The development of a reproducible model system for the study of Hepatitis C virus (HCV) infection has the potential to significantly enhance the study of virus-host interactions and provide future direction for modeling the pathogenesis of HCV. While there are studies describing global gene expression changes associated with HCV infection, changes in the proteome have not been characterized. We report the first large scale proteome analysis of the highly permissive Huh-7.5 cell line containing a full length HCV replicon. We detected > 4,400 proteins in this cell line, including HCV replicon proteins, using multidimensional liquid chromatographic (LC) separations coupled to mass spectrometry (MS). The set of Huh-7.5 proteins confidently identified is, to our knowledge, the most comprehensive yet reported for a human cell line. Consistent with the literature, a comparison of Huh-7.5 cells (+) and (-) the HCV replicon identified expression changes of proteins involved in lipid metabolism. We extended these analyses to liver biopsy material from HCV-infected patients where > 1,500 proteins were detected from 2 {micro}g protein lysate using the Huh-7.5 protein database and the accurate mass and time (AMT) tag strategy. These findings demonstrate the utility of multidimensional proteome analysis of the HCV replicon model system for assisting the determination of proteins/pathways affected by HCV infection. Our ability to extend these analyses to the highly complex proteome of small liver biopsies with limiting protein yields offers the unique opportunity to begin evaluating the clinical significance of protein expression changes associated with HCV infection.},
doi = {10.1128/JVI.79.12.7558-7569.2005},
journal = {Journal of Virology, 79(12):7558-7569},
issn = {0022-538X},
number = 12,
volume = 79,
place = {United States},
year = {2005},
month = {6}
}