Project 3: Genetic Analysis of JCPyV Entry

项目摘要--项目3 JCPyV是一种小型DNA病毒，感染了很大一部分人类人口。很少，在免疫抑制的情况下 在个体中，JCPyV从肾上皮细胞传播到中枢神经系统，在那里它可以引起 快速致命性脱髓鞘病，进行性多灶性白质脑病，PML。病毒和细胞 导致这种致病转变的因素尚不清楚，也没有特定的抗病毒药物可以 抑制JCPyV。JCPyV衣壳分解和细胞内转运的许多方面还知之甚少。 在功能基因组学、伪病毒的生产和使用以及病毒/细胞生物学方面的最新进展 关于其他多瘤病毒的进入，但我们对JCPyV的理解还没有取得重大进展 进入。在这个项目中，我们将应用这些技术来识别和表征细胞中的 JCPyV进入和细胞内贩运。这一努力是通过遗传病毒学和细胞 DiMaio实验室的生物学专业知识，与其他项目负责人Atwood博士和 斯泰尔。病毒侵入的细节可能是JCPyV生物学活性的分子基础 PML的发病机制。此外，对JCPyV进入的复杂过程的研究可能会揭示 潜在的新的抗病毒靶点，并提出了抑制JCPyV复制和传播的其他策略。我们 将测试对JCPyV感染的候选进入因素的要求，并表征这些因素的作用 因素，特别强调开发和部署新的分析方法，以确定病毒生命的哪个阶段 这些因素的作用是循环的。此外，在机械学研究中，我们将寻找 并对它们进行突变分析，以确定它们的关键特征。最后，如有必要， 我们将进行大规模的基因筛查，以确定JCPyV感染所需的额外细胞基因。 这些研究将为JCPyV感染提供机制上的洞察，并为新的 治疗方法。这些检测还将用于鉴定产生的JCPyV突变体的进入 Atwood博士和Stehle博士，并分析由这些项目开发的病毒抑制剂的效果。

PROJECT SUMMARY - PROJECT 3 JCPyV is a small DNA virus that infects a large fraction of the human population. Rarely, in immunosuppressed individuals, JCPyV spreads from kidney epithelial cells into the central nervous system, where it can cause a rapidly fatal demyelination disease, progressive multifocal leukoencephalopathy, PML. The viral and cellular factors responsible for this pathogenic switch are not known, and there are no specific anti-viral agents that inhibit JCPyV. Many aspects of JCPyV capsid disassembly and intracellular trafficking are poorly understood. Recent advances in functional genomics, pseudovirus production and use, and virus/cell biology have shed light on entry of other polyomaviruses, but major inroads have yet to be made in our understanding of JCPyV entry. In this project, we will apply these techniques to identify and characterize cellular proteins that mediate JCPyV entry and intracellular trafficking. This endeavor is made possible by the genetic virologic and cell biology expertise of the DiMaio laboratory, in collaboration with the other project leaders, Drs. Atwood and Stehle. The details of virus entry may underlie the molecular basis for the biological activity of JCPyV during the pathogenesis of PML. Furthermore, study of the complex process of JCPyV entry is likely to reveal potential new anti-viral targets and suggest additional strategies to inhibit JCPyV replication and spread. We will test the requirement for candidate entry factors in JCPyV infection and to characterize the role of these factors with a particular emphasis on developing and deploying new assays to determine where in the virus life cycle the factors act. In addition, in mechanistic studies, we will search for a physical association between the cellular factors and conduct mutational analysis on them to identify their crucial features. Finally, if necessary we will conduct large-scale genetic screens to identify additional cellular genes required for JCPyV infection. These studies will provide mechanistic insight into JCPyV infection and inform the development of new therapeutic approaches. These assays will also be used to characterize entry by the JCPyV mutants generated by Dr. Atwood and Dr. Stehle, and to analyze the effects of viral inhibitors developed by these projects.