Yi Zhigang, Fang Caiyun, Pan Tingting, Wang Jiadong, Yang Pengyuan, Yuan Zhenghong
Key Laboratory of Medical Molecular Virology, Institute of Medical Microbiology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Biochem Biophys Res Commun. 2006 Nov 10;350(1):174-8. doi: 10.1016/j.bbrc.2006.09.027. Epub 2006 Sep 18.
Hepatitis C virus (HCV) RNA synthesis takes place on a detergent resistant membrane (DRM) structure. To identify potential cellular proteins related to HCV replication complexes (RC), we purified DRMs from HCV subgenomic replicon cells and its parental Huh7 cells. The proteins of DRM fractions were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. Comparing with parental Huh7 cells, 60 proteins were up-regulated while 14 proteins were down-regulated in HCV replicon cells. Ras-GTPase-activating protein binding protein 1 (G3BP1), one of the elevated proteins, was found to be associated with HCV NS5B and knockdown of G3BP1 by siRNA in HCV replicon cells significantly reduced HCV replication, which may indicate it a potential component of HCV RC. These results suggest that HCV viral gene and proteins may regulate the presence of host cellular proteins in DRM, ensure appropriate concentrations of replication components, and hence control the rates or efficiencies of HCV replication.
丙型肝炎病毒（HCV）RNA合成发生在抗去污剂膜（DRM）结构上。为了鉴定与HCV复制复合物（RC）相关的潜在细胞蛋白，我们从HCV亚基因组复制子细胞及其亲本Huh7细胞中纯化了DRM。通过二维凝胶电泳分离DRM组分的蛋白质，并通过质谱鉴定。与亲本Huh7细胞相比，HCV复制子细胞中有60种蛋白质上调，14种蛋白质下调。上调的蛋白质之一Ras-GTPase激活蛋白结合蛋白1（G3BP1）被发现与HCV NS5B相关，并且在HCV复制子细胞中通过siRNA敲低G3BP1可显著降低HCV复制，这可能表明它是HCV RC的潜在组分。这些结果表明，HCV病毒基因和蛋白质可能调节DRM中宿主细胞蛋白的存在，确保复制组分的适当浓度，从而控制HCV复制的速率或效率。
