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Hepatitis C virus RNA polymerase and NS5A complex with a SNARE-like protein.
Tu, H; Gao, L; Shi, S T; Taylor, D R; Yang, T; Mircheff, A K; Wen, Y; Gorbalenya, A E; Hwang, S B; Lai, M M.
Afiliación
  • Tu H; Department of Molecular Microbiology and Immunology, University of Southern California School of Medicine, Los Angeles, California 90033, USA.
Virology ; 263(1): 30-41, 1999 Oct 10.
Article en En | MEDLINE | ID: mdl-10544080
Hepatitis C virus (HCV) NS5A is a phosphoprotein that possesses a cryptic trans-activation activity. To investigate its potential role in viral replication, we searched for the cellular proteins interacting with NS5A protein by yeast two-hybrid screening of a human hepatocyte cDNA library. We identified a newly discovered soluble N-ethylmaleimide-sensitive factor attachment protein receptor-like protein termed human vesicle-associated membrane protein-associated protein of 33 kDa (hVAP-33). In vitro binding assay and in vivo coimmunoprecipitation studies confirmed the interaction between hVAP-33 and NS5A. Interestingly, hVAP-33 was also shown to interact with NS5B, the viral RNA-dependent RNA polymerase. NS5A and NS5B bind to different domains of hVAP-33: NS5A binds to the C-terminus, whereas NS5B binds to the N-terminus of hVAP-33. Immunofluorescent staining showed a significant colocalization of hVAP-33 with both NS5A and NS5B proteins. hVAP-33 contains a coiled-coil domain followed by a membrane-spanning domain at its C-terminus. Cell fractionation analysis revealed that hVAP-33 is predominantly associated with the ER, the Golgi complex, and the prelysosomal membrane, consistent with its potential role in intracellular membrane trafficking. These interactions provide a mechanism for membrane association of the HCV RNA replication complex and further suggest that NS5A is a part of the viral RNA replication complex.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Polimerasa Dependiente del ARN / Proteínas Portadoras / Proteínas no Estructurales Virales / Hepacivirus / Proteínas de Transporte Vesicular / Proteínas de la Membrana Tipo de estudio: Prognostic_studies Límite: Animals / Female / Humans Idioma: En Revista: Virology Año: 1999 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Polimerasa Dependiente del ARN / Proteínas Portadoras / Proteínas no Estructurales Virales / Hepacivirus / Proteínas de Transporte Vesicular / Proteínas de la Membrana Tipo de estudio: Prognostic_studies Límite: Animals / Female / Humans Idioma: En Revista: Virology Año: 1999 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos