Use of synchrotron radiation to investigate the structure of genomic RNAs inside viral capsids

最简单的病毒仅由核酸和蛋白质分子的保护壳组成。我们正在研究核酸成分为单链RNA的病毒的结构、组装和拆卸。ssRNA病毒涵盖了广泛的感染因子，包括那些导致普通感冒、某些癌症和艾滋病的病原体。抵御任何病毒最成功的防线是接种适当的免疫原，但在许多情况下，这已被证明是非常困难或不可能的。我们相信，通过我们最近开展的工作，对ssRNA在这些病毒的组装和拆卸中的作用有了更好的理解，这给我们带来了独特的见解，将开辟新的治疗靶点，有可能使子代病毒组装的效率大大降低，从而使非传染性的、部分组装的颗粒可以启动宿主的免疫系统，从而导致病毒清除。这项工作的核心是对病毒颗粒内ssRNA结构的理解。因此，我们建议测试用于研究这种结构的主要技术，即x射线足迹，是否可以用于我们的测试病毒。这项工作涉及使用牛津郡的新钻石同步加速器，在那里我们将建立一个仪器，让病毒以一种确定的方式暴露在x射线下。由此产生的改良的无害的ssrna将在利兹和我们在俄克拉何马州的美国合作者进行检查。

The simplest viruses consist only of nucleic acids surrounded by a protective shell of protein molecules. We are studying the structures, assembly and disassembly of viruses in which the nucleic acid component is a single-stranded (ss) RNA. ssRNA viruses encompass broad classes of infectious agents including those responsible for the common cold, some cancers and AIDS. The most successful line of defence against any virus is vaccination with an appropriate immunogen but in many cases this has proved to be very difficult or impossible. We believe that the improved understanding of the roles of the ssRNA in the assembly and disassembly of such viruses resulting from the work we have recently carried out gives us unique insights that will open up novel therapeutic targets, potentially allowing the efficiency of progeny virus assembly to be reduced dramatically so that non-infectious, partially assembled particles can prime the host's immune system, leading to viral clearance. Central to this effort is an understanding of teh structure(s) of the ssRNA within the viral particle. We are therefore proposing to test whether the main technique available for interogating such structures, X-ray footprinting, can be used on our test viruses. This work involves use of the new Diamond Synchrotron in Oxfordshire where we will build an instrument to allow viruses to be exposed to X-rays in a defined way. The resulting modified and harmless ssRNAs will then be examined in Leeds and by our US collaborators in Oklahoma.