title: Association of filamin A and vimentin with hepatitis C virus proteins in infected human hepatocytes.
abstract: Chronic hepatitis C (CHC) infection caused by hepatitis C virus (HCV) is a major cause of liver disease and remains a major therapeutic challenge. A variety of host proteins interact with HCV proteins. The definitive role of cytoskeletal (CS) proteins in HCV infection remains to be determined. In this study, our aim was to determine the expression profile of differentially regulated and expressed selected CS proteins and their association with HCV proteins in infected hepatocytes as possible therapeutic targets. Using proteomics, qRT-PCR, Western blot and immunofluorescence techniques, we revealed that filamin A (fila) and vimentin (vim) were prominently increased proteins in HCV-expressing human hepatoma cells compared with parental cells and in liver biopsies from patients with CHC vs controls. HCV nonstructural (NS) 3 and NS5A proteins were associated with fila, while core protein partially with fila and vim. Immunoprecipitation showed interactions among fila and NS3 and NS5A proteins. Cells treated with interferon-α showed a dose- and time-dependent decrease in CS and HCV proteins. NS proteins clustered at the perinuclear region following cytochalasin b treatment, whereas disperse cytoplasmic and perinuclear distribution was observed in the no-treatment group. This study demonstrates and signifies that changes occur in the expression of CS proteins in HCV-infected hepatocytes and, for the first time, shows the up-regulation and interaction of fila with HCV proteins. Association between CS and HCV proteins may have implications in future design of CS protein-targeted therapy for the treatment for HCV infection.
authors.affiliation: The Liver-Biliary-Pancreatic Center and Liver, Digestive and Metabolic Disorders Laboratory, Carolinas Medical Center, Charlotte, NC 28203, USA. sriparna.ghosh@carolinashealthcare.org
authors.forename: S
authors.initials: S
authors.lastname: Ghosh
authors.forename: W A
authors.initials: WA
authors.lastname: Ahrens
authors.forename: S U
authors.initials: SU
authors.lastname: Phatak
authors.forename: S
authors.initials: S
authors.lastname: Hwang
authors.forename: L W
authors.initials: LW
authors.lastname: Schrum
authors.forename: H L
authors.initials: HL
authors.lastname: Bonkovsky
mesh_terms: D015153:Blotting, Western; D003285:Contractile Proteins / Q000378:metabolism*; D064448:Filamins; D005455:Fluorescent Antibody Technique; D016174:Hepacivirus / Q000472:pathogenicity*; D022781:Hepatocytes / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D008840:Microfilament Proteins / Q000378:metabolism*; D011485:Protein Binding; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome; D014746:Vimentin / Q000378:metabolism*; D014758:Viral Core Proteins / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
chemical_list: D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D020543:Proteome; D014746:Vimentin; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C069130:nucleocapsid protein, Hepatitis C virus; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase