权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanisms of viral protein trafficking in polarized insect cells

极化昆虫细胞中病毒蛋白运输的机制

基本信息

批准号：
2024252
负责人：
Gary Blissard
金额：
$ 77.99万
依托单位：
Boyce Thompson Institute Plant Research
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2024252&HistoricalAwards=false
关键词：
Mechanisms viral protein trafficking polarized

项目摘要

Arthropods represent the largest animal biomass on earth. Their populations are regulated by many viral diseases, and they vector many viruses that cause diseases of plants and animals. Typically, a virus enters the insect by the oral route and must penetrate the cells of the gut to spread to other tissues. Because little is known in this area, these studies will focus on the specific mechanisms by which viruses move through the insect gut cells. Typically, a virus will infect and replicate in the gut cells and newly-produced particles will bud from the other side, into the insect's blood. These studies will examine how the virus and its components move through the cell in a highly specific directional manner, so that the virus can enter the insect's blood and eventually infect other tissues. In another component of these studies, viral protein signals that are recognized by the cell, for recruitment into cellular trafficking pathways will be identified. These studies capitalize on the powerful genetic system in the fruitfly, Drosophila melanogaster. This system will be used in combination with cell culture systems and other modern molecular techniques, to identify the insect proteins and viral signals that are required for polarized virus trafficking through insect gut cells. The studies will have impacts across a wide variety of virus-insect interactions, with applications in diverse areas such as agricultural crop protection, forestry, animal science, and human health. In addition, undergraduate and graduate student student training will be supported by this proposal.Viral transport across the insect midgut epithelium is an important infection step for most viruses that infect insects, and for those viruses vectored by insects. However, this process is not well-understood even in the best characterized systems due to the difficult nature of experimental manipulation in the insect gut. These studies incorporate an experimental design that leverages two powerful model systems for identifying specific proteins, pathways, and signals involved in virus-midgut trafficking. Using ectopic expression of viral proteins in Drosophila midgut cells, followed by RNAi screens using comprehensive libraries of RNAi fly strains, candidate pathways will be screened to identify specific proteins and pathways necessary for polarized trafficking of two model viral proteins, baculovirus GP64 and VSV G. To identify viral protein signals or motifs necessary for midgut trafficking, the baculovirus GP64 protein will be analyzed in a natural host insect, the lepidopteran Trichoplusia ni. Existing libraries of viruses containing previously characterized envelope protein mutations will be used to identify the viral protein signals recognized for recruitment to trafficking pathways. The identification of viral signals and host midgut proteins and pathways for polarized trafficking will establish new paradigms for pathogen-insect interactions and will dramatically advance the understanding of how viruses productively interact with insects as hosts and vectors.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

节肢动物是地球上生物量最大的动物。它们的数量受到许多病毒性疾病的控制，它们携带许多导致动植物疾病的病毒。通常，病毒通过口腔途径进入昆虫，必须穿透肠道细胞传播到其他组织。由于在这一领域所知甚少，这些研究将集中在病毒通过昆虫肠道细胞的具体机制上。通常情况下，病毒会在肠道细胞中感染和复制，新产生的颗粒会从另一边发芽，进入昆虫的血液。这些研究将检查病毒及其成分如何以高度特定的定向方式在细胞中移动，以便病毒可以进入昆虫的血液并最终感染其他组织。在这些研究的另一个组成部分，病毒蛋白信号被细胞识别，招募到细胞运输途径将被确定。这些研究利用了果蝇（Drosophila melanogaster）强大的遗传系统。该系统将与细胞培养系统和其他现代分子技术结合使用，以识别极化病毒通过昆虫肠道细胞运输所需的昆虫蛋白质和病毒信号。这些研究将对各种各样的病毒-昆虫相互作用产生影响，并在农业作物保护、林业、动物科学和人类健康等不同领域得到应用。此外，本提案将支持本科生和研究生的学生培训。病毒通过昆虫中肠上皮的转运是大多数感染昆虫的病毒的一个重要感染步骤，对于那些由昆虫传播的病毒来说也是如此。然而，由于在昆虫肠道中进行实验操作的困难性质，即使在最好的表征系统中，这一过程也没有得到很好的理解。这些研究结合了一个实验设计，利用两个强大的模型系统来识别涉及病毒-中肠运输的特定蛋白质、途径和信号。利用果蝇中肠细胞中病毒蛋白的异位表达，然后利用RNAi蝇株综合文库进行RNAi筛选，筛选候选途径，以确定杆状病毒GP64和VSV g两种模型病毒蛋白极化运输所需的特定蛋白质和途径。为了确定中肠运输所需的病毒蛋白信号或基序，杆状病毒GP64蛋白将在天然宿主昆虫中进行分析。鳞翅目毛鳞目现有的包含先前表征的包膜蛋白突变的病毒文库将用于识别被贩运途径招募的病毒蛋白信号。病毒信号和宿主中肠蛋白以及极化运输途径的鉴定将为病原体-昆虫相互作用建立新的范例，并将极大地促进对病毒如何作为宿主和媒介与昆虫有效相互作用的理解。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。