Direct interaction between alpha-actinin and hepatitis C virus NS5B |
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Authors: | Lan Shuiyun Wang Hua Jiang Hong Mao Hongxia Liu Xiaoying Zhang Xiaonan Hu Yunwen Xiang Li Yuan Zhenghong |
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Affiliation: | Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai 200032, PR China. |
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Abstract: | It has been suggested that cellular proteins are involved in hepatitis C virus (HCV) RNA replication. By using the yeast two-hybrid system, we isolated seven cDNA clones encoding proteins interacting with HCV RNA polymerase (NS5B) from a human liver cDNA library. For one of these, alpha-actinin, we confirmed the interaction by coimmunoprecipitation, immunofluorescent staining and confocal microscopic analysis. Experiments with deletion mutants showed that domains NS5B(84-95), NS5B(466-478), and alpha-actinin(621-733) are responsible for the interaction. Studies of the HCV subgenomic replicon system with small interference RNA indicate that alpha-actinin is essential for HCV RNA replication. Our results suggest alpha-actinin may be a component of the HCV replication complex. |
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Keywords: | Hepatitis C Virus RNA polymerase α-Actinin Protein-protein interaction Yeast two-hybrid system |
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