Alpha-Herpesvirus Transport in Axons

描述(申请人提供)：阿尔法疱疹病毒是嗜神经的病原体，包括人类病毒水痘带状疱疹病毒(VZV)和单纯疱疹病毒(HSV)1和2，以及动物病毒伪狂犬病病毒(PRV)。这些病毒通过定向轴突运输进入神经系统，到达外周神经节的感觉神经元细胞体，在那里建立了终生潜伏感染。感染通常不会对健康人的生命造成威胁，然而，这些病毒能够跨突触传播，通过电路特有的途径进入大脑。一旦进入大脑，导致的脑炎在大多数情况下是致命的。我研究的长期目标是确定阿尔法疱疹病毒在神经系统中传播的机制，并确定控制疾病结局的因素。这项应用将专注于病毒在感觉神经元轴突中的传播。具体地说，就是： 使用感染性克隆诱变方法，将分离出携带编码每种被膜蛋白基因突变的病毒。 突变病毒将通过对融合到绿色荧光蛋白上的衣壳的延时跟踪来筛选轴突中衣壳运输的缺陷。 将启动衣壳运输所需的被膜蛋白的结构/功能分析。 通过这种方式，病毒被膜蛋白在衣壳运输中的作用将得到明确的测试。这些蛋白质有望将衣壳与微管马达捆绑在一起，并调节运动活动，因为目前还没有其他类型的病毒蛋白与衣壳结合，并同时暴露在宿主细胞质中。这些研究的结果将确定阿尔法疱疹病毒如何在感觉神经元内定向移动。这将是确定病毒传播的分子机制的第一步，并解决病毒在脊椎动物神经系统中传播的更大问题。

DESCRIPTION (provided by applicant): Alpha-herpesviruses are neurotropic pathogens that include the human viruses varicella-zoster virus (VZV) and herpes simplex virus (HSV) types 1 and 2, and the animal virus pseudorabies virus (PRV). These viruses spread into the nervous system by directed axonal transport to sensory neuron cell bodies in peripheral ganglia, where life-long latent infections are established. Infections are typically not life threatening in healthy individuals, however, these viruses have the capacity to spread trans-synaptically to enter the brain by circuit-specific routes. Once in the brain, the resulting encephalitis is lethal in the majority of cases. The long-term goals of my studies are to determine the mechanisms by which alpha-herpesviruses spread in the nervous system, and to identify the factors governing disease outcome. This application will focus on virus spread in the axons of sensory neurons. Specifically: Using infectious clone mutagenesis methods, viruses will be isolated carrying mutations in the genes encoding each tegument protein. The mutant viruses will be screened for defects in capsid transport in axons by time-lapse tracking of capsids fused to the green-fluorescence protein. Structure/function analysis of tegument proteins required for capsid transport will be initiated. In this way, the role of the viral tegument proteins in capsid transport will be definitively tested. These proteins are expected to both tether capsids to microtubule motors and regulate motor activity, as no other class of viral proteins are known to associate with capsids and be exposed to the host cytosol simultaneously. The results of these studies will determine how alpha-herpesviruses move directionally within sensory neurons. This will serve as a first step to identifying the molecular mechanisms of viral transport, and to addressing the larger question of virus spread in the vertebrate nervous system.