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

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.

α疱疹病毒亚科包括神经系统病原体，例如1型和2型单纯疱疹病毒（HSV-1和HSV-2）和猪病毒伪狂犬病病毒（PRV）。初始感染通常发生在粘膜上皮，例如口腔和肛门生殖器粘膜。对于 HSV，以及对于 PRV 的鼻和口咽粘膜，在这些组织中复制后，子代病毒颗粒被释放并感染末端。然后，它们通过微管引导的逆行交通沿着轴突到达神经细胞体，最终将病毒基因组传递到细胞核，并在随后的人类三叉神经节潜伏期中以环状双链 DNA 形式持续存在。 -1) 和猪 (PRV) 中，病毒基因表达、DNA 复制和新衣壳组装后会定期重新激活，并从细胞核中出现。成熟的α疱疹病毒颗粒从神经元细胞体转运到轴突并沿轴突转运，但人们对这些转运和包膜事件在α疱疹病毒感染的神经元中发生的方式和地点知之甚少。通过使用驱动蛋白马达的微管引导的顺行运输，感染性成熟病毒颗粒在神经末梢积聚，然后释放以感染邻近的粘膜。上皮细胞，导致随后几轮病毒复制和传播，每个阶段使用的驱动蛋白马达的身份，用于招募驱动蛋白来运输病毒体的机制，甚至沿着轴突运输的病毒颗粒的结构，无论是否有包膜。衣壳或在神经元细胞体中获得包膜的衣壳是我们在本应用中解决的关键问题，在具体目标 1 中，我们研究了 gE/gI-US9p 膜蛋白的功能。在将驱动蛋白马达 KIF1A 和 KIF5 募集到 HSV-1 和 PRV 过程中，我们还测试了一项关于大皮膜蛋白 UL36p 在将 KIF5 组装到运输病毒颗粒中的作用的创新假设。陷阱”来解决HSV-1包膜在何处获得它的有争议的问题，无论是在细胞体还是在神经末梢（分别是“结合”和“分离”机制）我们还测试了有关 HSV-1 和 PRV 通过“已婚”和“分离”途径流出期间驱动蛋白招募机制的关键问题。 α疱疹病毒从潜伏期重新激活后从神经系统传播到粘膜表面的主要事件：衣壳包膜、微管定向运输和顺行轴突具体目标是利用两位主要研究人员互补的体外和体内专业知识，以及我们对 gE/gI-US9p 复合物和 UL36p 的共同兴趣和经验，来剖析支持这些病毒引起的复发性疾病的分子机制。