Mechanisms in Polyomavirus Assembly

描述（由申请人提供）：病毒学中的一个新兴概念是病毒复制和病毒体组装通常在细胞内称为病毒“工厂”的位点的特定位置机械偶联。这些过程的空间和时间协调允许以能量上有利的方式装配线生产病毒体，同时允许最终结构的质量控制。尽管在一些地方已经描述了工厂 关于病毒在细胞质中复制的细节，大多数DNA病毒在细胞核中复制/组装，其中由于密集的染色质网络，工厂的形态特征难以识别和表征。也有迹象表明，这些工厂侵占了重要的核结构域，否则这些核结构域将用于DNA损伤修复（DDR）等功能，而且事实上，在DNA病毒的复制位点发现了许多参与DDR的细胞蛋白。我们建议采用病毒和宿主细胞遗传学、生化分离和高分辨率成像技术相结合的方法，来表征病毒粒子组装工厂的结构和蛋白质组成。我们的目标是以纳米分辨率确定这些工厂的分子结构，并确定其蛋白质组分的空间关系。随着最近鉴定出9种新的人类多瘤病毒，以及用于多种疾病的免疫调节疗法的显著增加，多瘤病毒已成为目前尚无特异性疗法的重要人类病原体。组装工厂的表征不仅将确定影响多瘤病毒复制以及其他DNA病毒的新治疗靶点，而且还提供了关于它们在正常细胞过程中的功能的见解。

DESCRIPTION (provided by applicant): 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 protein composition of virion assembly factories for the well-studied murine polyomavirus, using a combined approach employing virus and host cell genetics, biochemical isolation, and high-resolution imaging technologies. Our goal is to determine the molecular architecture of these factories at nanometer resolution, and determine the spatial relationship of their protein components. With the recent identification of nine 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.