National Research Laboratory of Hepatitis C Virus and Ilsong Institute of Life Science, Hallym University, Anyang, South Korea.
Institut Pasteur Korea, Bundang-gu, Seongnam, Republic of Korea.
Sci Rep. 2017 Jul 25;7(1):6445. doi: 10.1038/s41598-017-06740-9.
The life cycle of hepatitis C virus (HCV) is highly dependent on host proteins for virus propagation. By transcriptome sequencing analysis, we identified host genes that were highly differentially expressed in HCV-infected cells. Of these candidates, we selected Death receptor 6 (DR6) for further characterization. DR6 is an orphan member of the tumor necrosis factor receptor superfamily. In the present study, we demonstrated that both mRNA and protein levels of DR6 were increased in the context of HCV replication. We further showed that promoter activity of DR6 was increased by HCV infection. By employing promoter-linked reporter assay, we showed that HCV upregulated DR6 via ROS-mediated NF-κB pathway. Both mRNA and protein levels of DR6 were increased by NS4B or NS5A. However, NS5A but not NS4B specifically interacted with DR6. We showed that HCV modulated JNK, p38 MAPK, STAT3, and Akt signaling pathways in a DR6-dependent manner. Interestingly, Akt signaling cascade was regulated by protein interplay between DR6 and NS5A. Silencing of DR6 expression resulted in decrease of infectious HCV production without affecting viral entry, replication, and translation. Together, these data indicate that HCV modulates DR6 signaling pathway for viral propagation and may contribute to HCV-mediated pathogenesis.
丙型肝炎病毒 (HCV) 的生命周期高度依赖于宿主蛋白来进行病毒繁殖。通过转录组测序分析,我们鉴定了在 HCV 感染细胞中高度差异表达的宿主基因。在这些候选基因中,我们选择了死亡受体 6 (DR6) 进行进一步表征。DR6 是肿瘤坏死因子受体超家族的孤儿成员。在本研究中,我们证明了在 HCV 复制的情况下,DR6 的 mRNA 和蛋白水平均增加。我们进一步表明,HCV 感染可增加 DR6 的启动子活性。通过启动子连接报告基因检测,我们表明 HCV 通过 ROS 介导的 NF-κB 途径上调 DR6。DR6 的 mRNA 和蛋白水平均被 NS4B 或 NS5A 增加。然而,NS5A 而不是 NS4B 特异性地与 DR6 相互作用。我们表明,HCV 以 DR6 依赖的方式调节 JNK、p38 MAPK、STAT3 和 Akt 信号通路。有趣的是,Akt 信号级联反应受 DR6 和 NS5A 之间的蛋白相互作用调节。沉默 DR6 的表达会导致感染性 HCV 产量减少,而不影响病毒进入、复制和翻译。总之,这些数据表明 HCV 调节 DR6 信号通路以促进病毒繁殖,并可能导致 HCV 介导的发病机制。