CMV Control of Host Membrane Trafficking

CMV 对宿主膜运输的控制

• 批准号: 9982022
• 负责人: Felicia D Goodrum
• 金额: $ 4.13万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982022
• 关键词: Address; Affect; Aging; Biogenesis; Biology; Cell Fractionation; Cells; Cellular Membrane; Cellular biology; Complex; Cytomegalovirus; Cytomegalovirus Infections; DNA Viruses; Disease; Endothelial Cells; Environment; Exhibits; Exocytosis; Failure; Family; Genes; Goals; Herpesviridae; Human; Immunosuppression; Infection; Inflammation; Integration Host Factors; Intracellular Membranes; Mediating; Membrane; Morphology; Multivesicular Body; Mutation; Organelles; Pathway interactions; Phenotype; Positioning Attribute; Process; Proteins; Proteomics; RNA Interference; RNA Viruses; Recruitment Activity; Regulation; Role; Signal Transduction; Site; Transplantation; Vesicle; Viral; Viral Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; congenital infection; design; extracellular; high resolution imaging; insight; intercellular communication; knock-down; member; recombinant virus; success; trafficking; virus host interaction

PROJECT SUMMARY / ABSTRACT Viruses commandeer host membrane trafficking to promote viral replication. While this is a critical control point for virus infection, how viruses hijack host proteins and pathways to promote infection is incompletely understood. Viral-mediated alterations in membrane trafficking are best understood for building replication compartments and sites of assembly for RNA viruses. However, much less is known about how DNA viruses influence membrane remodeling in the host. Human cytomegalovirus (CMV) is a complex DNA virus and a member of the herpesvirus family that establishes a lifelong infection in the host. During infection, CMV induces alterations in membrane trafficking that results in increased biogenesis of multivesicular bodies (MVBs), organelles important for signaling and exocytosis or degradation of cargo. During infection, MVBs become loaded with virus particles and dense bodies (vesicles of viral tegument protein). The mechanisms by which CMV controls host membrane trafficking pathways, and particularly MVB biogenesis, are not understood. Therefore, defining the viral proteins and host targets important for the regulation of membrane trafficking is an important goal with important implications for host and virus biology. We have identified two CMV proteins, pUL135 and pUL136, which regulate MVB biogenesis. Recombinant viruses lacking UL135 or UL136 genes exhibit profound alterations in MVB biogenesis and the incorporation of viral cargo into MVB. Consistent with these phenotypes, we have identified host interacting proteins for both pUL135 and pUL136 proteins that are important for membrane trafficking and MVB biogenesis. The discovery of two viral proteins and their host interactions strongly position us to define the mechanisms by which CMV modulates membrane trafficking and MVB biogenesis. We hypothesize that UL135- and UL136-host interactions co-opt host membrane trafficking and MVB pathways to modulate virus replication. Three aims are proposed to address this hypothesis. We will define the association of UL135 and UL136 with subcellular membrane compartments and host proteins in Aim 1. In Aim 2, we will define the significance of host interactions to virus replication, membrane trafficking, and incorporation of cargo into MVBs to understand the mechanistic basis of CMV- mediated control of host trafficking. Finally, we will investigate the role of UL135 and UL136 and their host interacting partners in regulating MVB biogenesis and determine how this regulation impacts viral egress. Our studies will define the virus-host interactions and trafficking pathways targeted by CMV and their significance to infection. Furthermore, we anticipate that our findings will uncover common strategies used by many viruses to manipulate trafficking pathways.

项目概要/摘要 病毒征用宿主膜运输以促进病毒复制。虽然这是一个关键控制点 对于病毒感染来说，病毒如何劫持宿主蛋白和途径来促进感染尚不完全 明白了。病毒介导的膜运输改变对于构建复制来说是最好的理解 RNA 病毒的区室和组装位点。然而，人们对 DNA 病毒如何 影响宿主的膜重塑。人类巨细胞病毒 (CMV) 是一种复杂的 DNA 病毒， 疱疹病毒家族的成员，可在宿主体内形成终生感染。感染期间，巨细胞病毒 诱导膜运输的改变，导致多囊泡体的生物发生增加 （MVB），对于信号传导和胞吐作用或货物降解很重要的细胞器。感染期间，MVB 充满病毒颗粒和致密体（病毒外皮蛋白的囊泡）。其机制由 其中 CMV 控制宿主膜运输途径，特别是 MVB 生物发生，但不是 明白了。因此，定义病毒蛋白和宿主靶标对于膜的调节很重要 贩运是一个重要目标，对宿主和病毒生物学具有重要影响。我们已经确定了两个 CMV 蛋白 pUL135 和 pUL136，调节 MVB 生物发生。缺乏 UL135 的重组病毒或 UL136 基因在 MVB 生物发生和病毒货物掺入 MVB 中表现出深刻的改变。 与这些表型一致，我们鉴定了 pUL135 和 pUL136 的宿主相互作用蛋白 对膜运输和 MVB 生物发生很重要的蛋白质。两种病毒蛋白的发现 及其宿主相互作用使我们能够明确 CMV 调节膜的机制 贩运和 MVB 生物发生。我们假设 UL135 和 UL136 宿主相互作用增选宿主 膜运输和 MVB 途径来调节病毒复制。提出了三个目标来解决 这个假设。我们将定义 UL135 和 UL136 与亚细胞膜区室的关联 目标 1 中的宿主蛋白。在目标 2 中，我们将定义宿主相互作用对病毒复制的重要性， 膜运输，以及将货物掺入 MVB 中，以了解 CMV 的机制基础 对宿主贩运的中介控制。最后，我们将研究 UL135 和 UL136 及其宿主的作用 相互作用的合作伙伴调节 MVB 生物合成，并确定这种调节如何影响病毒的流出。我们的 研究将确定 CMV 靶向的病毒-宿主相互作用和贩运途径及其对 感染。此外，我们预计我们的发现将揭示许多病毒使用的常见策略 操纵贩运途径。