Zhou Hui, Qian Qi, Shu Ting, Xu Jiuyue, Kong Jing, Mu Jingfang, Qiu Yang, Zhou Xi
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, 430071, China.
Virol Sin. 2020 Aug;35(4):436-444. doi: 10.1007/s12250-019-00182-5. Epub 2019 Nov 27.
RNAi interference (RNAi) is an evolutionarily conserved post-transcriptional gene silencing mechanism and has been well recognized as an important antiviral immunity in eukaryotes. Numerous viruses have been shown to encode viral suppressors of RNAi (VSRs) to antagonize antiviral RNAi. Hepatitis C virus (HCV) is a medically important human pathogen that causes acute and chronic hepatitis. In this study, we screened all the nonstructural proteins of HCV and found that HCV NS2 could suppress RNAi induced either by small hairpin RNAs (shRNAs) or small interfering RNAs (siRNAs) in mammalian cells. Moreover, we demonstrated that NS2 could suppress RNAi via its direct interaction with double-stranded RNAs (dsRNAs) and siRNAs, and further identified that the cysteine 184 of NS2 is required for the RNAi suppression activity through a serial of point mutation analyses. Together, our findings uncovered that HCV NS2 can act as a VSR in vitro, thereby providing novel insights into the life cycle and virus-host interactions of HCV.
RNA干扰(RNAi)是一种进化上保守的转录后基因沉默机制,并且已被公认为真核生物中重要的抗病毒免疫机制。大量病毒已被证明可编码RNAi病毒抑制因子(VSR)以对抗抗病毒RNAi。丙型肝炎病毒(HCV)是一种具有医学重要性的人类病原体,可导致急性和慢性肝炎。在本研究中,我们筛选了HCV的所有非结构蛋白,发现HCV NS2可在哺乳动物细胞中抑制由小发夹RNA(shRNA)或小干扰RNA(siRNA)诱导的RNAi。此外,我们证明NS2可通过其与双链RNA(dsRNA)和siRNA的直接相互作用来抑制RNAi,并通过一系列点突变分析进一步确定NS2的半胱氨酸184是RNAi抑制活性所必需的。总之,我们的研究结果揭示HCV NS2在体外可作为一种VSR,从而为HCV的生命周期和病毒-宿主相互作用提供了新的见解。