Tanji Y, Hijikata M, Hirowatari Y, Shimotohno K
Virology Division, National Cancer Center Research Institute, Tokyo, Japan.
J Virol. 1994 Dec;68(12):8418-22. doi: 10.1128/JVI.68.12.8418-8422.1994.
Hepatitis C virus (HCV) serine proteinase (Cpro-2) is responsible for the processing of HCV nonstructural (NS) protein processing. To clarify the mechanism of Cpro-2-dependent processing, pulse-chase and mutation analyses were performed by using a transient protein production system in cultured cells. Pulse-chase study revealed the sequential production of HCV-NS proteins. Production of p70(NS3) and p66(NS5B) were rapid. An 89-kDa processing intermediate protein (p89) was observed during the early part of the chase. p89 seemed to be cleaved first into a 31-kDa protein (p31) and a p58/56(NS5A). p31 was further processed into p4(NS4A) and p27(NS4B). Mutation analysis of cleavage sites of NS4A/4B, NS4B/5A, and NS5A/5B revealed that cleavage at each site is essentially independent from cleavage occurring at the other site.
丙型肝炎病毒（HCV）丝氨酸蛋白酶（Cpro-2）负责HCV非结构（NS）蛋白的加工。为阐明Cpro-2依赖性加工的机制，利用培养细胞中的瞬时蛋白产生系统进行了脉冲追踪和突变分析。脉冲追踪研究揭示了HCV-NS蛋白的顺序产生。p70（NS3）和p66（NS5B）的产生迅速。在追踪早期观察到一种89 kDa的加工中间蛋白（p89）。p89似乎首先被切割成一种31 kDa的蛋白（p31）和一种p58/56（NS5A）。p31进一步被加工成p4（NS4A）和p27（NS4B）。对NS4A/4B、NS4B/5A和NS5A/5B切割位点的突变分析表明，每个位点的切割基本上独立于其他位点的切割。
