Interplay of RNA structure, protein and miRNA binding in early replication events of hepatitis C virus

丙型肝炎病毒（HCV）是一种重要的病毒性病原体，感染了全球约3%的人口。丙肝病毒感染可导致慢性肝病、肝硬化和肝细胞癌。目前的抗病毒疗法效果不佳，而且开发出预防感染的疫苗的前景渺茫。开发新的抗病毒疗法面临两个重大挑战；首先，我们对病毒如何控制其生命周期中的关键事件只有不完全的了解，特别是在感染的早期阶段，当时只有低病毒水平。其次，病毒复制会产生大量的变异后代病毒，其中包括对抗病毒药物有抗性的变异。我们已经确定了细胞蛋白和病毒核酸之间的一种新的相互作用。抑制这种相互作用？通过阻止蛋白质与病毒基因组结合？显著降低病毒复制的能力。这是通过抑制生命周期中称为翻译的关键早期过程来实现的。这种抑制的结果是病毒产量降低。我们预测？基于我们自己和别人的工作？结合蛋白将病毒基因组稳定在一个特定的构象中破坏这个构象对病毒复制是有害的。为了更好地理解这一过程，我们建议进行三组实验；1)我们将测量病毒基因组结构的变化，以确认它是否经历了我们预测的结构变化；2)我们将识别蛋白质并开发试剂，使我们能够确定它是如何起作用的？例如，我们将测试它是否只存在于肝细胞中，我们将研究它是否在未感染和病毒感染的细胞中发生变化（我们预测会发生变化），无论是数量、位置还是通过化学修饰。3)我们将开展功能研究，研究该蛋白对病毒生命周期的重要性。例如，我们将耗尽细胞中的蛋白质，看看这是否会减少病毒的产量，具体来说，它是否会减少病毒生命周期中的关键转译事件。所有的病毒都是寄生虫，它们破坏细胞过程，有利于自己的复制。如果正如我们预测的那样，HCV的基因组结构通过？一种来自肝细胞的细胞蛋白，那么抑制这一过程可能允许开发新的，有效的抗病毒治疗方法。

Hepatitis C virus (HCV) is an important viral pathogen infecting approximately 3% of the global population. HCV infection results in chronic liver disease, cirrhosis and hepatocellular carcinoma. Current anti-viral therapies are poorly effective and there is little prospect of developing a vaccine to prevent infection. There are two significant challenges to the development of new anti-viral therapies; firstly we have only an incomplete understanding of how the virus controls critical events in its life cycle, particularly during the early stages of infection when there are only low viral levels. Secondly, virus replication generates large populations of variant progeny viruses, these include variants resistant to anti-viral drugs. We have identified a novel interaction between a cellular protein and the nucleic acid of the virus. Inhibition of this interaction ? by preventing the protein from binding to the virus genome ? significantly reduces the ability of the virus to replicate. This is achieved by inhibiting a key early process in the life cycle termed translation. The result of this inhibition is that the viral yield is reduced. We predict ? based on our own work and that of others ? that the binding protein stabilizes the virus genome in a particular conformation and that disrupting this conformation is detrimental for virus replication. To understand this process better we propose to conduct three sets of experiments; 1) we will measure changes in the structure of the virus genome to confirm whether it undergoes the structural changes we predict, 2) we will identify the protein and develop reagents that will allow us to determine how it functions ? for example, we will test if it only present in liver cells, we will investigate whether it changes in uninfected and virus infected cells (we predict it does), either in quantity, location or by chemical modification. 3) we will conduct functional studies to investigate the importance of this protein to the virus life cycle. For example, we will deplete the protein from cells and see if this reduces the yield of virus and, specifically, whether it reduces the key translation events in the virus life cycle. All viruses are parasites that subvert cellular processes to favour their own replication. If, as we predict, the genomic structure of HCV is somehow stabilized by ?stealing? a cellular protein from liver cells, then inhibition of this process may allow the development of novel, effective anti-viral therapies.