Neurotropic herpesvirus envelopment and microtubule-mediated transport

嗜神经性疱疹病毒包膜和微管介导的运输

基本信息

批准号：
10585954
负责人：
Gregory Allan Smith
金额：
$ 67.07万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-15 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10585954
关键词：
Address Afferent Neurons Alphaherpesvirinae Axon Axonal Transport Bacterial Artificial Chromosomes Binding Biochemistry Blindness Capsid Cell Line Cell Nucleus Cells Cellular biology Complex Congenital Abnormality Cytoplasmic Organelle DNA biosynthesis Development Disease Episome Epithelium Event Family suidae Fluorescence Microscopy Gene Expression Goals Herpes Labialis Herpesviridae Herpesvirus 1 Human Human Herpesvirus 2 Image In Vitro Induced pluripotent stem cell derived neurons Infection Kinesin Laboratories Mediating Membrane Proteins Microtubules Molecular Motor Mucous Membrane Nerve Nervous System Neurons Nose Oral Organelles Oropharyngeal Pathogenesis Pathway interactions Periodicals Principal Investigator Process Proteins Role Simplexvirus Structure Structure of trigeminal ganglion Surface Testing Tissues Travel Viral Viral Genes Viral Genome Virion Virus Virus Replication anterograde transport ds-DNA experience human disease in vivo induced pluripotent stem cell innovation interest mad itch virus neuronal cell body neuronal transport neurotropic novel novel therapeutics particle pathogen protein complex reactivation from latency recruit trafficking transmission process

Address Afferent Neurons Alphaherpesvirinae Axon Axonal Transport Bacterial Artificial Chromosomes Binding Biochemistry Blindness Capsid Cell Line Cell Nucleus Cells Cellular biology Complex Congenital Abnormality Cytoplasmic Organelle DNA biosynthesis Development Disease Episome Epithelium Event Family suidae Fluorescence Microscopy Gene Expression Goals Herpes Labialis Herpesviridae Herpesvirus 1 Human Human Herpesvirus 2 Image In Vitro Induced pluripotent stem cell derived neurons Infection Kinesin Laboratories Mediating Membrane Proteins Microtubules Molecular Motor Mucous Membrane Nerve Nervous System Neurons Nose Oral Organelles Oropharyngeal Pathogenesis Pathway interactions Periodicals Principal Investigator Process Proteins Role Simplexvirus Structure Structure of trigeminal ganglion Surface Testing Tissues Travel Viral Viral Genes Viral Genome Virion Virus Virus Replication anterograde transport ds-DNA experience human disease in vivo induced pluripotent stem cell innovation interest mad itch virus neuronal cell body neuronal transport neurotropic novel novel therapeutics particle pathogen protein complex reactivation from latency recruit trafficking transmission process

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项目摘要

The Alphaherpesvirinae include pathogens of the nervous system such as herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) and the swine virus pseudorabies virus (PRV). Initial infection is commonly at mucosal epithelia such as the oral and anogenital mucosa for HSV, and nasal and oropharyngeal mucosa for PRV. Following replication in these tissues, progeny viral particles are released and infect the termini of adjacent sensory neurons. They then travel by microtubule-directed retrograde traffic along the axon to the neuronal cell body. The viral genome is ultimately delivered to the nucleus and persists as a circular dsDNA episome during ensuing latency in the trigeminal ganglia of humans (HSV-1) and swine (PRV). Periodic reactivation from latency is followed by viral gene expression, DNA replication and assembly of new capsids. These become packaged with the viral genome, emerge from the nucleus and bud into cytoplasmic organelles to generate enveloped, infectious viral particles in the organellar lumen. How and where these transport and envelopment events occur in alphaherpesvirus-infected neurons is poorly understood. Maturing alphaherpesvirus particles are transported from the neuronal cell body into and along the axon by microtubule-directed anterograde transport using kinesin motors. Infectious mature viral particles accumulate at the nerve terminal then are released to infect adjacent mucosal epithelia, leading to subsequent rounds of viral replication and spread. The identity of the kinesin motors utilized at each stage, the machinery used to recruit kinesins to trafficking virions and even the structure of the viral particle that traffics down the axon, whether non-enveloped capsids or capsids that have acquired envelopes in the neuronal cell body, are key questions that we address in this application. In Specific Aim 1 we investigate the function of the gE/gI-US9p membrane protein complex in recruitment of the kinesin motors KIF1A and KIF5 to HSV-1 and PRV. We also test an innovative hypothesis concerning the role of the large tegument protein UL36p in assembly of KIF5 onto trafficking virions. In Specific Aim 2 we use a novel “envelopment trap” to address the controversial question of where HSV-1 acquires its envelope, whether in the cell body or nerve terminal (the Married and Separate mechanisms, respectively) in a range of neuronal cell lines, human iPSC-derived Trigeminal Ganglia and explanted sensory neurons. We also test key questions concerning the mechanisms of kinesin recruitment during egress of HSV-1 and PRV via the Married and Separate pathways. This proposal is therefore focused on the three major events that underlie alphaherpesvirus transmission from the nervous system to mucosal surfaces following reactivation from latency: capsid envelopment, microtubule-directed trafficking, and anterograde axonal transport. The specific aims exploit the complementary in vitro and in vivo expertise of the two principal investigators, and our common interests and experience with the gE/gI-US9p complex and UL36p, to dissect the molecular mechanisms supporting recrudescent disease caused by these viruses.

Alphaherpesvirinae包括神经系统的病原体，例如单纯疱疹病毒类型1和2（HSV-1和HSV-2）和猪病毒假疗法病毒（PRV）。初始感染通常是在粘膜上皮中的，例如HSV的口服和肛门粘膜，以及PRV的鼻和口咽粘膜。在这些组织中复制后，释放后代病毒颗粒并感染了相邻感觉神经元的末端。然后，他们沿着微管定向的逆行交通沿轴突传播到神经元细胞体。在增强人类三叉神经节的潜伏期（HSV-1）和猪（PRV）的潜伏期期间，病毒基因组最终被传递到细胞核中，并作为圆形DSDNA发作持续存在。潜伏期的周期性重新激活之后是病毒基因表达，DNA复制和新衣壳的组装。这些被病毒基因组包装，从细胞核和芽中出现到细胞质细胞器中，以产生有机腔中的包膜，传染性的病毒颗粒。这些转运和包膜发生在αHerpesvirus感染的神经元中的方式和何处发生在较少的了解中。使用驱动蛋白电动机通过微管定向的顺行传输将成熟的αHERPESVIRUS颗粒从神经元细胞体转移到轴突中。然后释放出在神经末端积累的传染性成熟病毒颗粒以感染相邻的粘膜上皮，从而导致随后的病毒复制和扩散。在每个阶段使用的驱动蛋白电动机的身份，用于募集驱动蛋白的机械，甚至是流动轴突的病毒颗粒的结构，无论是在神经元细胞体中获得的包膜，无论是非发育的capsids还是非发育的衣壳，都是我们在该应用程序中解决的关键问题。在特定目标1中，我们研究了GE/GI-US9P膜蛋白复合物在募集驱动蛋白电机KIF1A和KIF5对HSV-1和PRV中的功能。我们还检验了关于大型远程蛋白UL36P在KIF5组装到运输病毒中的作用的创新假设。在特定目标2中，我们使用一种新颖的“信封陷阱”来解决HSV-1在何处获得其包膜的有争议的问题，无论是在细胞体还是神经末端（分别是已婚和单独的机制，分别为已婚机制），在一系列神经元细胞系，人IPSC源自IPSC的三叉神经节和expantented的感官神经元中。我们还通过已婚和单独的途径测试了有关HSV-1和PRV出口期间驱动蛋白招募机制的关键问题。因此，该提案的重点是从潜伏期重新激活后，从神经系统传播到粘膜表面的三个主要事件：衣壳环境，微管定向的贩运和抗反式轴突运输。该特定目的探讨了两个主要研究者的体外和体内专业知识，以及我们对GE/GI-US9P复合物和UL36P的共同利益和经验，以剖析支持这些病毒引起的重新产生疾病的分子机制。