Molecular and cellular mechanisms of HSV-1 assembly and egress

项目摘要/摘要 阿尔法疱疹病毒，包括人类重要的病原体单纯疱疹病毒1型(HSV-1)，是 在为数不多的进化到利用高度专业化的神经细胞生物学的病毒中。在.期间 自然病程，阿尔法疱疹病毒感染周围神经系统(PNS)神经节，作为一种 终生潜伏感染，偶尔重新激活，导致周围组织复发，或者可以 传播到感染中枢神经系统。在重新激活、复制和组装时，子代病毒 颗粒可以从受感染神经元的胞体/树突中出来，或者可以分类成轴突并经历长时间的 距离轴突运输。特别是HSV-1，是病毒性脑炎的主要原因，也可能是 神经退行性疾病的发展。 本补充方案的主要目的是确定以运动为基础的微管 调节轴突分类的机制，特别是在三叉神经节神经元中。在目标1中，我们将确定 不同的动蛋白马达和微管相关蛋白对子病毒颗粒细胞内转运的影响 在非神经细胞中，通过活细胞成像。在目标2中，我们将确定这些马达和 微流体样小室在初级神经元轴突分选中的应用 神经元培养系统。在本补充申请中，建议的实验是新颖的，但在 上级R01奖励的范围。 阐明我们的病毒在神经元和非神经元中使用的基本细胞生物学过程 细胞将增加我们对疱疹病毒如何以及为什么传播到神经系统并在神经系统内传播的理解， 导致确定可用药靶点和开发更好的病毒神经病理学治疗方法，以及 可能提供对细胞生物学的基本见解，特别是对神经元的细胞生物学。

Project Summary/Abstract Alpha herpesviruses, including the important human pathogen Herpes Simplex Virus 1 (HSV-1), are among the very few viruses that have evolved to exploit highly-specialized neuronal cell biology. During the natural course of disease, alpha herpesviruses infect peripheral nervous system (PNS) ganglia, persist as a life-long latent infection, and occasionally reactivate to cause recurrent lesions in peripheral tissues, or can spread to infect the central nervous system. Upon reactivation, replication, and assembly, progeny virus particles can exit from the soma/dendrites of infected neurons, or can sort into axons and undergo long- distance axonal transport. HSV-1, in particular, is a leading cause of viral encephalitis, and may also contribute to the development of neurodegenerative disease. The main objective of this supplement proposal is to determine the microtubule motor-based mechanisms that mediate axonal sorting, specifically in PNS neurons. In Aim 1, we will determine the roles of different kinesin motors and microtubule-associated proteins in intracellular transport of progeny virus particles in non-neuronal cells by live-cell imaging. In Aim 2, we will determine the roles of these motors and microtubule-associated proteins in axonal sorting in primary neurons, using a microfluidics-like chambered neuronal culture system. In this supplement application, the proposed experiments are novel, but are within the scope of the parent R01 award. Elucidating the basic cell biological processes that our viruses use in both neurons and non-neuronal cells will increase our understanding of how and why herpesviruses spread to and within the nervous system, lead to the identification of druggable targets and development of better therapies for viral neuropathology, and may provide fundamental insights into cell biology, particularly of the cell biology of neurons.