title: Hepatitis C virus NS5A protein impairs TNF-mediated hepatic apoptosis, but not by an anti-FAS antibody, in transgenic mice.
abstract: Hepatitis C virus (HCV) is a major etiologic agent of chronic hepatitis worldwide and may lead to the development of hepatocellular carcinoma. However, the mechanism of development of chronic hepatitis or hepatocarcinogenesis by HCV remains unclear. In the present study, we have investigated the effect of nonstructural protein 5A (NS5A) on TNF- and Fas-mediated apoptosis in the liver of transgenic mice. For this purpose, transgenic mice were generated by targeting the HCV NS5A genomic region cloned under the control of a liver-specific apoE promoter. The transgenic animals were phenotypically similar to their normal littermates and did not exhibit a detectable histological change in the liver at 8-12 weeks of age. Intraperitoneal injection of recombinant TNF induced hepatic injury and apoptosis in normal mice. In contrast, transgenic mice expressing NS5A protein were protected against hepatic apoptosis after injection of TNF. However, injection of anti-Fas antibody into transgenic mice did not significantly influence hepatic apoptosis compared to the normal littermates. These results suggested distinct effects of TNF and anti-Fas antibody in transgenic mice expressing NS5A. We subsequently investigated the effect of NS5A in signaling pathways involved in these two cytokine-mediated apoptosis. A physical association between NS5A and TRADD was observed by pull-down assay, coimmunoprecipitation, and colocalization experiments. Furthermore, NS5A prevented the association between TRADD and FADD and blocked TRADD-mediated NF-kappaB activation. Together, our results suggest that NS5A impairs TNF-mediated apoptosis by interfering upstream of the signal transduction pathway and may play a role in HCV-mediated pathogenesis.
authors.affiliation: Department of Pathology, Saint Louis University, 1402 S Grand Blvd., 4th Floor, St. Louis, MO 63104, USA.
authors.forename: Mainak
authors.initials: M
authors.lastname: Majumder
authors.forename: Asish K
authors.initials: AK
authors.lastname: Ghosh
authors.forename: Robert
authors.initials: R
authors.lastname: Steele
authors.forename: Xiao Yan
authors.initials: XY
authors.lastname: Zhou
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Phillips
authors.forename: Ranjit
authors.initials: R
authors.lastname: Ray
authors.forename: Ratna B
authors.initials: RB
authors.lastname: Ray
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D017209:Apoptosis; D002352:Carrier Proteins / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein; D005260:Female; D006801:Humans; D008099:Liver / Q000166:cytology / Q000473:pathology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011506:Proteins / Q000378:metabolism; D053199:TNF Receptor-Associated Death Domain Protein; D047990:TNF Receptor-Associated Factor 1; D014162:Transfection; D014407:Tumor Cells, Cultured; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D019014:fas Receptor / Q000276:immunology*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000906:Antibodies; D002352:Carrier Proteins; D053200:Fas-Associated Death Domain Protein; D011506:Proteins; D053199:TNF Receptor-Associated Death Domain Protein; D047990:TNF Receptor-Associated Factor 1; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins; D014409:Tumor Necrosis Factor-alpha; D017361:Viral Nonstructural Proteins; D019014:fas Receptor; C505533:FADD protein, human; C505534:Fadd protein, mouse; C481226:Tradd protein, mouse; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase