Mechanisms in Polyomavirus Assembly

项目总结 病毒学中的一个新概念是病毒复制和病毒粒子组装通常是机械性的 结合在细胞内特定位置的称为病毒“工厂”的位置。空间和时间 这些过程的协调允许装配线以以下方式生产病毒粒子 在考虑到最终结构的质量控制的同时，在能量上有利。尽管工厂有 对在细胞质中复制的病毒进行了更详细的描述，大多数DNA病毒 在核内复制/组装，在那里工厂的形态特征难以识别 并且由于染色质网络致密而具有特征性。也有迹象表明，这些工厂 篡夺其他用于DNA损伤修复(DDR)等功能的基本核结构域， 事实上，在DNA病毒的复制位置发现了许多参与DDR的细胞蛋白质。我们 建议对病毒粒子组装厂的结构和组成进行表征，以供深入研究 用iPOND蛋白鉴定联合方法鉴定小鼠多瘤病毒 分辨率成像和病毒/宿主细胞遗传学。我们的目标是确定分子的结构 这些工厂的纳米分辨率以及它们的空间和功能关系 蛋白质成分。最近发现了10种新的人类多瘤病毒，并标记了 随着对各种疾病免疫调节疗法的增加，多瘤病毒已经成为 目前还没有特效药的重要人类病原体。刻画人物形象 组装厂不仅将确定影响多瘤病毒复制的新治疗靶点， 以及其他DNA病毒，但也提供了对它们在正常细胞过程中的功能的洞察。

PROJECT SUMMARY An emerging concept in virology is that viral replication and virion assembly are often mechanistically coupled in specific locations within the cell at sites termed virus “factories”. The spatial and temporal coordination of these processes permit assembly line production of virions in a manner that is energetically favorable while allowing for quality control of the final structure. Although factories have been described in some detail for viruses replicating in the cytoplasm, most DNA viruses replicate/assemble in the nucleus where morphological features of factories are problematic to identify and characterize due to the dense chromatin network. There are also indications that the factories usurp essential nuclear domains otherwise used for such functions as DNA damage repair (DDR), and indeed many cellular proteins involved in DDR are found at replication sites for DNA viruses. We propose to characterize the structure and composition of virion assembly factories for the well-studied murine polyomavirus using a combined approach employing iPOND protein identification, high resolution imaging, and virus/host cell genetics. Our goal is to determine the molecular architecture of these factories at nanometer resolution as well as the spatial and functional relationships of their protein components. With the recent identification of ten new human polyomaviruses, and the marked increase in immunomodulatory therapies for a variety of diseases, polyomaviruses have emerged as important human pathogens that at present have no specific therapy. Characterization of the assembly factories not only will identify new therapeutic targets for affecting polyomavirus replication, as well as other DNA viruses, but also provide insight as to their function in normal cellular processes.