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

RNA 结构、蛋白质和 miRNA 结合在丙型肝炎病毒早期复制事件中的相互作用

• 批准号: G1100139/1
• 负责人: David Evans
• 金额: $ 59.91万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=G1100139%2F1
• 关键词: Interplay; RNA; structure; protein; miRNA

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.

丙型肝炎病毒（HCV）是一种重要的病毒病原体，感染全球约3%的人口。HCV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，并且开发预防感染的疫苗的前景很小。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，病毒复制产生大量变异子代病毒，这些包括对抗病毒药物具有抗性的变异体。我们已经确定了一种新的细胞蛋白质和病毒核酸之间的相互作用。抑制这种相互作用？通过阻止蛋白质与病毒基因组结合会显著降低病毒的复制能力这是通过抑制生命周期中一个关键的早期过程来实现的，称为翻译。这种抑制的结果是病毒产量降低。我们预测？基于我们自己和其他人的研究该结合蛋白使病毒基因组稳定在特定的构象中，并且破坏这种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）我们将鉴定蛋白质并开发试剂，使我们能够确定它是如何发挥作用的？例如，我们将测试它是否只存在于肝细胞中，我们将调查它是否在未感染和病毒感染的细胞中发生变化（我们预测它会发生变化），无论是在数量上，位置上还是通过化学修饰。3)我们会进行功能研究，探讨这种蛋白质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，特别是，它是否会减少病毒生命周期中的关键翻译事件。所有的病毒都是寄生虫，破坏细胞过程以利于自身复制。如果像我们预测的那样，HCV的基因组结构在某种程度上是稳定的？偷窃？一种来自肝细胞的细胞蛋白，那么抑制这一过程可能会允许开发新的、有效的抗病毒疗法。