Viral and Cellular Determinants of Hepatitis C Virus RNA Replication in Cell Culture

摘要: Studies on the replication of hepatitis C virus (HCV) have been facilitated by development selectable subgenomic replicons replicating in human hepatoma cell line Huh-7 at a surprisingly high level. Analysis replicon population selected cells revealed occurrence culture-adaptive mutations that enhance RNA substantially. To gain better understanding HCV culture adaptation, we characterized conserved identified sequence analysis 26 independent clones for their effect replication. Mutations enhancing were found nearly every nonstructural (NS) protein, and they could be subdivided into least two groups efficiency cooperativity: (i) NS3 with low impact but enhanced cooperatively when combined highly adaptive (ii) NS4B, -5A, -5B, causing strong increase being incompatible each other. In addition to mutations, host plays an equally important role efficient We tested several passages same up 100-fold differences ability support amplification. These not due variations internal ribosome entry site-dependent translation or degradation. search cellular factor(s) might responsible different levels permissiveness cells, decreased increasing amounts transfected RNA, indicating viral proteins are cytopathic factors limit summary, these data show is determined both adaptation itself.