Subversion of lysosomal trafficking by HHV-7 U21

HHV-7 U21 颠覆溶酶体贩运

• 批准号: 9706283
• 负责人: AMY W HUDSON
• 金额: $ 12.65万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9706283
• 关键词: Adaptive Immune System; Administrative Supplement; Affinity; Age; Binding; Biological; Biological Assay; Biological Process; CRISPR library; CRISPR/Cas technology; Cell surface; Cells; Cellular biology; Centrifugation; Client; Co-Immunoprecipitations; Complex; Comprehension; Coupled; Cytoplasmic Tail; Data; Detection; Evolution; Genetic Screening; Glycoproteins; Goals; Golgi Apparatus; Herpesviridae; Histocompatibility Antigens Class I; Host Defense Mechanism; Human Herpesvirus 6; Human Herpesvirus 7; Immune; Immune Evasion; Immune response; In Vitro; Integral Membrane Protein; Lead; Libraries; Life; Ligands; Lighting; Lysosomes; Mediating; Microscope; Microscopy; Mitochondria; Molecular; Mutate; Pathogenesis; Pathway interactions; Physiologic pulse; Play; Population; Prevention; Process; Proteins; RNA Interference; Role; Route; Signal Transduction; Sorting - Cell Movement; Structure; Textbooks; Transmembrane Domain; Viral Proteins; Virus; experimental study; genome-wide; in vivo; instrument; mutant; nanomolar; novel; protein transport; proteostasis; small hairpin RNA; success; symposium; trafficking

PROJECT SUMMARY/ABSTRACT. The goal of this administrative supplement request is to provide a state-of-the- art microscope for cell biological studies of lysosomal sorting for R01GM120735. The requested instrument will be shared equally with Drs. Blake Hill (R01GM067180.) and Matt Scaglione (R35GM119544) for their cell biological studies in mitochondria fission and proteostasis, respectively. The herpesviruses have co-evolved with their hosts for hundreds of millions of years. As our adaptive immune systems evolved, so did the mechanisms of immune evasion encoded by herpesviruses. Because herpesviruses remain latent or persistent within their host throughout life, the herpesviruses have evolved a unique set of strategies to help them contend with the lifelong host immune response. This application’s central premise is that the herpesviruses, in their evolution, have performed a 100 million-year-long genetic screen to undermine host defense mechanisms, and in so doing, can illuminate cell biological processes that we may not yet understand; we seek to take advantage of their evolution to discover new concepts in cell biology. The long term goal of this project is to understand the underlying cell biological mechanisms by which human herpesviruses-6 and -7 (HHV-6/7) escape host defense mechanisms. The focus of this proposal is to understand how HHV-7 U21 reroutes class I MHC molecules to lysosomes, and in so doing, understand the cellular pathway usurped by U21. Our preliminary data indicate that U21 employs an unknown sorting mechanism that seems to defy convention - almost certainly utilizing a novel cellular means of lysosomal sorting. In Aim 1, we assess the structure and association of U21 with itself, as a homotetramers, and determine whether homooligomerization of U21 is necessary its ability to bind to and reroute class I MHC molecules to lysosomes, and use RUSH to examine the synchronous trafficking of U21 and class I molecules to lysosomes. We ultimately seek a temporal and mechanistic view of how U21 undermines the cellular lysosomal trafficking pathway, and how this lysosomal trafficking pathway functions in the absence of U21. A better understanding of this process should contribute to our understanding of basic underlying principles of lysosomal trafficking, and also lead to a more thorough comprehension of how these viruses have such great success in evading the host immune response.

项目摘要/摘要。此行政补充请求的目标是提供最新的- R01GM120735溶酶体分选细胞生物学研究的ART显微镜。所请求的仪器将 与布莱克·希尔博士(R01GM067180)平分。和Matt Scaglione(R35GM119544)用于他们的细胞 分别在线粒体分裂和蛋白稳定方面进行生物学研究。 疱疹病毒与宿主共同进化了数亿年。作为我们的适应性免疫 系统进化了，疱疹病毒编码的免疫逃避机制也进化了。因为 疱疹病毒在整个生命周期中在宿主内保持潜伏或持续，疱疹病毒已经进化成 一套独特的策略，帮助它们应对终生的宿主免疫反应。此应用程序的中心 前提是疱疹病毒在它们的进化过程中进行了长达1亿年的基因筛选 破坏宿主防御机制，在这样做的过程中，可以阐明我们可能无法做到的细胞生物学过程 但要理解；我们寻求利用它们的进化来发现细胞生物学的新概念。这个 该项目的长期目标是了解人类 疱疹病毒-6和-7(HHV-6/7)逃避宿主防御机制。这项提议的重点是 了解HHV-7 U21如何将I类MHC分子重新路由到溶酶体，并在这样做的过程中，了解 细胞通路被U21篡改。我们的初步数据表明，U21采用了一种未知的排序方式 似乎违反惯例的机制--几乎可以肯定地使用一种新的溶酶体的细胞方法 分类。在目标1中，我们评估了U21作为同源异构体的结构和与自身的联系，以及 确定U21的同源齐聚是否是必要的，其与I类MHC结合和重路由的能力 分子到溶酶体，并用HASH检测U21和I类分子的同步运输 到溶酶体。我们最终寻求U21如何破坏细胞的时间和机械观点 溶酶体转运途径，以及在没有U21的情况下溶酶体转运途径是如何发挥作用的。一个 更好地理解这一过程应该有助于我们理解 溶酶体的贩运，也有助于更彻底地理解这些病毒是如何具有如此巨大的 成功地逃避了宿主的免疫反应。