Cellular cofactors affecting hepatitis C virus infection and replication
Cellular cofactors affecting hepatitis C virus infection and replication
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DOI：
10.1073/pnas.0704894104
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发表时间：
2007-07-31
影响因子：
11.1
通讯作者：
Rice, Charles M.
中科院分区：
文献类型：
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作者：
Randall, Glenn;Panis, Maryline;Rice, Charles M.
最近鉴定的在细胞培养中具有感染性的丙型肝炎病毒（HCV）分离株提供了一个遗传系统来评估病毒-宿主相互作用对HCV复制的意义。我们已经完成了一个系统的RNAi筛选，其中siRNA被设计为靶向62个宿主基因，这些基因编码与HCV RNA或蛋白质物理相互作用的蛋白质，或者属于被认为调节HCV感染的细胞途径。这包括我们在本研究中鉴定的10种结合HCV NS 5A的宿主蛋白。靶向这些宿主基因中的26个的siRNA使感染性HCV产生改变>3倍。在这组26个siRNA中包括靶向Dicer的siRNA，Dicer是RNAi沉默途径的主要组分。与RNAi是哺乳动物中的抗病毒途径的假设相反，如已经报道的亚基因组HCV复制子，靶向Dicer的siRNA抑制HCV复制。此外，靶向RNAi途径的几种其他组分的siRNA也抑制HCV复制。人肝脏、人肝癌Huh-7.5细胞和携带复制型HCV的Huh-7.5细胞的microRNA谱表明，miR-122是每种环境中的主要microRNA。miR-122先前已涉及正调控HCV基因型1复制子的复制。我们发现miR-122的2 '-O-甲基反义寡核苷酸缺失也抑制HCV基因型2a复制和感染性病毒产生。我们的数据定义了26个调节HCV感染的宿主基因，并表明对HCV复制的功能性RNAi的需求比该途径可能对HCV产生的任何抗病毒活性都占主导地位。
Recently identified hepatitis C virus (HCV) isolates that are infectious in cell culture provide a genetic system to evaluate the significance of virus-host interactions for HCV replication. We have completed a systematic RNAi screen wherein siRNAs were designed that target 62 host genes encoding proteins that physically interact with HCV RNA or proteins or belong to cellular pathways thought to modulate HCV infection. This includes 10 host proteins that we identify in this study to bind HCV NS5A. siRNAs that target 26 of these host genes alter infectious HCV production >3-fold. included in this set of 26 were siRNAs that target Dicer, a principal component of the RNAi silencing pathway. Contrary to the hypothesis that RNAi is an antiviral pathway in mammals, as has been reported for sub-genomic HCV replicons, siRNAs that target Dicer inhibited HCV replication. Furthermore, siRNAs that target several other components of the RNAi pathway also inhibit HCV replication. MicroRNA profiling of human liver, human hepatoma Huh-7.5 cells, and Huh-7.5 cells that harbor replicating HCV demonstrated that miR-122 is the predominant microRNA in each environment. miR-122 has been previously implicated in positively regulating the replication of HCV genotype 1 replicons. We find that 2'-O-methyl antisense oligonucleotide depletion of miR-122 also inhibits HCV genotype 2a replication and infectious virus production. Our data define 26 host genes that modulate HCV infection and indicate that the requirement for functional RNAi for HCV replication is dominant over any antiviral activity this pathway may exert against HCV.