Equipe 4271, Université de Nantes, Paris, France.
J Virol. 2013 Jun;87(12):6668-77. doi: 10.1128/JVI.00538-13. Epub 2013 Apr 3.
In the model of Huh-7.5.1 hepatocyte cells infected by the JFH1 hepatitis C virus (HCV) strain, transcriptomic and proteomic studies have revealed modulations of pathways governing mainly apoptosis and cell cycling. Differences between transcriptomic and proteomic studies pointed to regulations occurring at the posttranscriptional level, including the control of mRNA translation. In this study, we investigated at the genome-wide level the translational regulation occurring during HCV infection. Sucrose gradient ultracentrifugation followed by microarray analysis was used to identify translationally regulated mRNAs (mRNAs associated with ribosomes) from JFH1-infected and uninfected Huh-7.5.1 cells. Translationally regulated mRNAs were found to correspond to genes enriched in specific pathways, including vesicular transport and posttranscriptional regulation. Interestingly, the strongest translational regulation was found for mRNAs encoding proteins involved in pre-mRNA splicing, mRNA translation, and protein folding. Strikingly, these pathways were not previously identified, through transcriptomic studies, as being modulated following HCV infection. Importantly, the observed changes in host mRNA translation were directly due to HCV replication rather than to HCV entry, since they were not observed in JFH1-infected Huh-7.5.1 cells treated with a potent HCV NS3 protease inhibitor. Overall, this study highlights the need to consider, beyond transcriptomic or proteomic studies, the modulation of host mRNA translation as an important aspect of HCV infection.
在受 JFH1 丙型肝炎病毒(HCV)株感染的 Huh-7.5.1 肝细胞模型中,转录组学和蛋白质组学研究揭示了主要调控细胞凋亡和细胞周期的途径的调节。转录组学和蛋白质组学研究之间的差异表明,调节发生在转录后水平,包括对 mRNA 翻译的控制。在这项研究中,我们在 HCV 感染过程中,从全基因组水平上研究了翻译调节。使用蔗糖梯度超速离心和微阵列分析来鉴定来自 JFH1 感染和未感染 Huh-7.5.1 细胞的翻译调节的 mRNA(与核糖体相关的 mRNA)。发现翻译调节的 mRNA 对应于富含特定途径的基因,包括囊泡运输和转录后调控。有趣的是,最强的翻译调节发生在编码参与前体 mRNA 剪接、mRNA 翻译和蛋白质折叠的蛋白质的 mRNAs 上。值得注意的是,这些途径以前没有通过 HCV 感染后的转录组学研究来确定。重要的是,观察到的宿主 mRNA 翻译的变化直接归因于 HCV 的复制,而不是 HCV 的进入,因为在用有效的 HCV NS3 蛋白酶抑制剂处理的 JFH1 感染的 Huh-7.5.1 细胞中没有观察到这些变化。总的来说,这项研究强调了在 HCV 感染中,除了转录组学或蛋白质组学研究之外,还需要考虑宿主 mRNA 翻译的调节,这是一个重要的方面。