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.

Alphaherpesvirine包括神经简单的单纯形病毒类型1和2（HSV-1和HSV-2）ABIES病毒（PRV）。对于这些组织在相邻的感觉神经元中的复制。三叉神经节的HSV-1）和猪（prv）。。在每个阶段使用的驱动蛋白电动机的身份神经元细胞体中的信封在募集动力素电机KIF1A和KIF5对HSV-1和PRV的募集中我们使用一个新颖的“信封陷阱”来解决HSV-1在哪里获得信封的有争议的问题，无论是在细胞体还是神经终端（已婚和单独的机制，各自的机制，分别），在一系列神经元细胞lins中。 Nterograde轴突运输。特定的目的是利用两个主要研究者的启示，我们的共同利益并体验了GE/GI-US9P Comp Complex和UL36P，以支持支持支持支持疾病的支持疾病的杀伤机制。