Direct interaction between α‐actinin and hepatitis C virus NS5B

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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, α‐actinin, we confirmed the interaction by coimmunoprecipitation, immunofluorescent staining and confocal microscopic analysis. Experiments with deletion mutants showed that domains NS5B84–95, NS5B466–478, and α‐actinin621–733 are responsible for the interaction. Studies of the HCV subgenomic replicon system with small interference RNA indicate that α‐actinin is essential for HCV RNA replication. Our results suggest α‐actinin may be a component of the HCV replication complex.