Structure and dynamics of enterovirus IRES-ribosome complexes

肠道病毒 IRES-核糖体复合物的结构和动力学

• 批准号: 2752057
• 金额: --
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2752057
• 关键词: Structure; dynamics; enterovirus; IRES; ribosome

Enteroviruses (e.g. Poliovirus, Rhinovirus, Coxsackievirus) comprise a diverse group of human and animal pathogens, which together are estimated to cause over one billion infections annually. Once inside the host cell, their positive-sense, single-stranded RNA genomes are directly translated to produce the viral polyprotein. A key event in the infection cycle is the recruitment of host ribosomes to initiate translation. This occurs via an internal ribosome entry site (IRES) in the 5' untranslated region of the genome. The enterovirus type 1 IRES is ~450 nt in length and organized into five structured domains, which interact with trans-acting protein factors (ITAFs) leading to recruitment of 43S complexes. However, our understanding of this network of interactions has been hampered by the lack of any high-resolution three-dimensional structures. Furthermore, a new gene has recently been discovered in some enteroviruses, located upstream of the main polyprotein (Lulla et al., Nat Microbiol, 2019). This exploits an alternate AUG codon located within domain VI of the IRES itself, but the mechanistic basis for start-site selection is not understood. You will study the structural and mechanistic basis of enterovirus initiation at a variety of model IRESs. This will involve the reconstitution of initiation in vitro, purification of initiation complexes, sample optimization, cryo-EM data collection and processing. You will also study protein RNA-interactions between ITAFs and IRES domains using a variety of biophysical and biochemical tools. You will explore key findings in virus-infected cells, in collaboration with the Lulla lab (University of Cambridge). You will join a vibrant, diverse and highly supportive training environment with the combined expertise of three supervisors - ribosome structural biology and protein-RNA interactions (Hill and Plevin labs) and Enterovirus molecular biology (Lulla lab). You will also benefit from access to state-of the art cryo-EM infrastructure at YSBL, and the molecular interactions laboratory at the Biology Technology Facility. Candidates from under-represented groups are particularly encouraged to apply.

肠道病毒（例如脊髓灰质炎病毒、鼻病毒、柯萨奇病毒）由多种人类和动物病原体组成，估计每年导致超过十亿例感染。一旦进入宿主细胞，它们的正义单链RNA基因组就会直接翻译以产生病毒多蛋白。感染周期中的一个关键事件是招募宿主核糖体来启动翻译。这是通过基因组 5' 非翻译区的内部核糖体进入位点 (IRES) 发生的。 1 型肠道病毒 IRES 长度约为 450 nt，分为五个结构域，这些结构域与反式作用蛋白因子 (ITAF) 相互作用，导致 43S 复合物的募集。然而，由于缺乏任何高分辨率的三维结构，我们对这种相互作用网络的理解受到了阻碍。此外，最近在一些肠道病毒中发现了一个新基因，位于主要多蛋白的上游（Lulla 等人，Nat Microbiol，2019）。这利用了位于 IRES 本身的结构域 VI 内的替代 AUG 密码子，但起始位点选择的机制基础尚不清楚。您将研究各种 IRES 模型中肠道病毒启动的结构和机制基础。这将涉及体外启动的重建、启动复合物的纯化、样品优化、冷冻电镜数据收集和处理。您还将使用各种生物物理和生化工具研究 ITAF 和 IRES 结构域之间的蛋白质 RNA 相互作用。您将与 Lulla 实验室（剑桥大学）合作，探索病毒感染细胞的重要发现。您将加入一个充满活力、多样化和高度支持的培训环境，并拥有三位导师的综合专业知识——核糖体结构生物学和蛋白质-RNA 相互作用（Hill 和 Plevin 实验室）以及肠道病毒分子生物学（Lulla 实验室）。您还将受益于 YSBL 最先进的冷冻电镜基础设施和生物技术设施的分子相互作用实验室。特别鼓励来自代表性不足群体的候选人申请。