Hepatitis C virus (HCV) NS5A binds RNA-dependent RNA polymerase (RdRP) NS5B and modulates RNA-dependent RNA polymerase activity.

摘要: Hepatitis C virus (HCV) NS5B is RNA-dependent RNA polymerase (RdRP), the essential catalytic enzyme for HCV replication. Recently, NS5A has been reported to be important establishment of replication in vitro by adaptive mutations, although its role viral remains uncertain. Here we report that purified bacterial recombinant and directly interact with each other vitro, detected glutathione S-transferase (GST) pull-down assay. Furthermore, complex formation these proteins transiently coexpressed mammalian cells was coprecipitation. Using terminally internally truncated NS5A, two discontinuous regions (amino acids 105–162 277–334) outside mutations were identified independently binding both vivo vitro(Yamashita, T., Kaneko, S., Shirota, Y., Qin, W., Nomura, Kobayashi, K., Mkyrakami, S. (1998) J. Biol. Chem. 273, 15479–15486). We previously examined effect His-NS5A on RdRP activity soluble NS5Bt vitro(see Yamashita et al. above). Wild weakly stimulated at first (when less than 0.1 molar ratio NS5B) then inhibited a dose-dependent manner. The internal deletion mutants defective exhibited no inhibitory effect, indicating necessary inhibition. Taken together, our results support idea modulates as component complex.