Determine the Three-Dimensional Structure of Icosahedral Virus Genomes

描述(申请人提供)：基因组是病毒的传染性物质，因此适当的包装、储存和拆卸是成功感染的关键。然而，人们对病毒基因组的三维结构知之甚少。我们开发了一种方法来确定包装在球形(二十面体)衣壳内的病毒基因组的结构。这项技术使用了低温电子显微镜的图像，可以三维重建基因组结构。该方法适用于模拟的图像数据，我们建议用卫星烟草花叶病毒(STMV)的图像来测试该技术，这是一个简单的模型系统。成功的3D重建依赖于对多个粒子图像进行平均，以及三维基因组排列从粒子到粒子保持一致的前提。我们希望将我们的研究扩展到微小核糖核酸病毒，这是一种重要的人类病原体家族，使用脊髓灰质炎病毒作为我们的模型系统。我们假设单个病毒物种的基因组具有一致的3D结构。通过验证这一假设，我们建议测试微小核糖核酸病毒用于我们的基因组重建方法的可行性。与公共卫生相关：为了有效感染，病毒的基因组必须在三个维度上正确排列。了解3D基因组结构将给出病毒如何导致疾病的更全面的机械图景，并可能为抗病毒疗法提供见解。

DESCRIPTION (provided by applicant): The genome is a virus' infectious material, thus proper packaging, storage, and disassembly are critical to successful infection. Yet, little is known about the three- dimensional structure of viral genomes. We developed a method for determining the structure of viral genomes that are packaged inside spherical (icosahedral) capsids. The technique uses images from cryogenic electron microscopy and allows three- dimensional reconstruction of the genome structure. The method works for simulated image data, and we propose to test the technique with images of satellite tobacco mosaic virus (STMV), a simple model system. Successful 3D reconstruction relies on averaging many particle images and the premise that the three-dimensional genome arrangement is consistent from particle-to-particle. We expect to extend our study to picornaviruses, a family of significant human pathogens, using poliovirus as our model system. We hypothesize that the genomes of individual viral species have a consistent 3D structure. By testing this hypothesis, we propose to test the feasibility of picornaviruses for our genome-reconstruction method. PUBLIC HEALTH RELEVANCE: For effective infection, a virus' genome must be arranged correctly in three dimensions. Understanding 3D genome architecture will give a fuller mechanistic picture of how viruses cause disease and may give insights into anti-viral therapies.