Isoform-specific interaction of pyruvate kinase with hepatitis C virus NS5B

Hepatitis C virus (HCV) replication and pathogenesis involve both virus-encoded proteins and cellular factors. In our study, we showed that NS5B, the HCV RNA-dependent RNA polymerase, interacted with M2 type pyruvate kinase (M2PK) but not L type pyruvate kinase. We confirmed the interaction by GST pull down, coimmunoprecipitation and confocal immunofluorescence analysis in cells with transient expression of NS5B and M2PK as well as in a HCV replicon-bearing cell line. Furthermore shRNA which specifically down-regulated M2PK expression could inhibit the replication of HCV in HCV replicon 9B cells.Structured summaryMINT-6548876, MINT-6548924:NS5B(Con1) (uniprotkb:Q9WMX2) and M2PK (uniprotkb:P14618) colocalize (MI:0403) by fluorescence microscopy (MI:0416)MINT-6548705:NS5BΔ21(BK) (uniprotkb:P26663) physically interacts (MI:0218) with M2PK (uniprotkb:P52480) by two hybrid (MI:0018)MINT-6548722, MINT-6548746:CM2PK (uniprotkb:P52480)binds (MI:0407) to NS5BΔ21(BK) (uniprotkb:P26663) by pull down (MI:0096)MINT-6548831:M2PK (uniprotkb:P14618) physically interacts (MI:0218) with NS5B (Con1) (uniprotkb:Q9WMX2) by anti bait coimmunoprecipitation (MI:0006)MINT-6548795:M2PK (uniprotkb:P14618) physically interacts (MI:0218) with NS5B(Con1) (uniprotkb:Q9WMX2)by anti tag coimmunoprecipitation (MI:0007)MINT-6548852:M2PK (uniprotkb:P14618) and NS5BΔ21(BK) (uniprotkb:P26663) colocalize (MI:0403) by fluorescence microscopy (MI:0416)