The roles of acidic autophagosomes in production of infectious poliovirus

酸性自噬体在传染性脊髓灰质炎病毒产生中的作用

• 批准号: 8630813
• 负责人: William T Jackson
• 金额: $ 32.26万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8630813
• 关键词: Appearance; Autophagocytosis; Autophagosome; Capsid Proteins; Cell membrane; Cells; Cellular Membrane; Cellular Stress; Characteristics; Cytosol; DNA Sequence Rearrangement; Data; Development; Encapsulated; Environment; Enzymes; Extracellular Space; Family; Family Picornaviridae; Future; Generations; Genome; Genomics; Goals; Homeostasis; Human; Human poliovirus; Infection; Lead; Life Cycle Stages; Lysosomes; Membrane; Modeling; Monitor; Morphogenesis; Nature; Organelles; Outcome; Pathway interactions; Play; Polioviruses; Process; Production; Proliferating; Proteins; Proteome; Proteomics; RNA; RNA Viruses; RNA replication; Research; Role; Signal Transduction; Site; Staging; Structure; Testing; Therapeutic; Therapeutic Agents; Time; Transport Vesicles; Vesicle; Viral; Virion; Virus; Virus Diseases; Virus Replication; Work; biological adaptation to stress; human disease; innovation; insight; novel; pathogen; phosphatidylinositol 4-phosphate; prevent; public health relevance; research study; therapeutic target; viral RNA

PROJECT SUMMARY/ABSTRACT All positive strand RNA viruses studied to date rearranged cellular membranes to promote their own replication. One reason for these rearrangements is that these viruses replicate their genomic RNA in association with cellular membranes. In poliovirus (PV), a model for a host of medically important positive strand RNA viruses, two distinct classes of vesicle have been identified. One class, a single-membraned vesicle which resembles a COPII secretory transport vesicle, associates with viral RNA replication proteins. The second type of vesicle, which is double-membraned, also associates with viral RNA replication proteins and resembles the autophagosome, an organelle induced by a pathway of cellular homeostasis and stress- response known as autophagy. Autophagosome-like vesicles are specifically induced by viruses and promote PV production. The long term goal of this project is to understand the mechanisms and consequences of cellular membrane rearrangements by picornaviruses. The objective of this application is to identify the roles played by autophagosomes during infection and define the mechanism by which PV induces autophagosome morphogenesis. Our preliminary data indicate that autophagy is required for optimal levels of infectious virus. Autophagic signaling is required for viral RNA replication. However, vesicle acidification, which in the case of autophagosomes is required for fusion with lysosomes, is required for cleavage of a viral capsid protein, the final step in generating infectious virus from newly formed virions. The central hypothesis of this proposal is that PV infection generates autophagosomes through morophological changes, including invagination, in the PIP4-rich secretory pathway-derived vesicles used for RNA replication. Acidification of the newly formed autophagosomes promotes virion maturation. The central hypothesis will be tested by pursuing two specific aims. In Aim I we will study the nature of the vesicle environment, and the requirements for virion maturation. In Aim II we will analyze the development of autophagic vesicles during infection and define the proteome of virus-induced autophagosomes. The rationale for this research is to understand how picornaviruses subvert what is often an anti-pathogen pathway to promote virion maturation. This will provide us with information needed to target this late step in virus production with therapeutics. Our innovative approaches will identify the mechanisms PV uses to induce autophagosomes, and how they promote maturation and egress of infectious virus. The proposed research is significant because it will fundamentally advance our understanding of how a medically important family of viruses subverts a basic cellular pathway to promote virus replication. This work will provide novel insights into the late stages of the viral life cycle, especially maturation and cellular egress, and provide the first steps in identifying therapeutic targets that may ultimately lead to treatments against multiple viral diseases. This work is also an important step in understanding the mechanisms of existing and future therapeutic agents against multiple viral diseases.

项目总结/摘要 迄今为止研究的所有正链RNA病毒都重排了细胞膜，以促进其自身的 复制的这些重排的一个原因是这些病毒复制它们的基因组RNA， 与细胞膜结合。在脊髓灰质炎病毒（PV）中，一种具有医学重要性的阳性 链RNA病毒，已经鉴定了两种不同类型囊泡。一类，单膜 类似于COPII分泌转运囊泡的囊泡，与病毒RNA复制蛋白相关。 第二种囊泡是双膜囊泡，也与病毒RNA复制蛋白有关 并且类似于自噬体，一种由细胞稳态和应激途径诱导的细胞器， 这种反应被称为自噬。自噬体样囊泡是由病毒特异性诱导的， 光伏生产。该项目的长期目标是了解 小核糖核酸病毒引起的细胞膜重排。此应用程序的目标是确定角色 自噬体在感染过程中发挥的作用，并定义PV诱导自噬体的机制 形态发生我们的初步数据表明，自噬是感染性病毒的最佳水平所必需的。 自噬信号是病毒RNA复制所必需的。然而，囊泡酸化，在这种情况下， 自噬体是与溶酶体融合所必需的，是切割病毒衣壳蛋白所必需的， 从新形成的病毒体产生感染性病毒的最后一步。这一提议的核心假设是 PV感染通过形态学变化产生自噬体，包括内陷， 用于RNA复制的富含PIP 4的分泌途径衍生的囊泡。酸化新形成的 自噬体促进病毒体成熟。中心假设将通过追求两个具体的 目标。在目的一，我们将研究囊泡环境的性质，和病毒粒子成熟的要求。 在目的II中，我们将分析感染过程中自噬囊泡的发育，并定义自噬囊泡的蛋白质组。 病毒诱导的自噬体。这项研究的基本原理是了解小核糖核酸病毒如何颠覆 这通常是一种抗病原体的途径来促进病毒体成熟。这将为我们提供信息 需要针对病毒产生的最后一步进行治疗。我们的创新方法将确定 PV用于诱导自噬体的机制，以及它们如何促进感染性细胞的成熟和排出。 病毒这项拟议中的研究意义重大，因为它将从根本上推进我们对人类如何在 医学上重要的病毒家族破坏了促进病毒复制的基本细胞途径。这项工作 将为病毒生命周期的后期阶段提供新的见解，特别是成熟和细胞出口， 并提供了确定治疗靶点的第一步，这些靶点可能最终导致治疗 多种病毒性疾病这项工作也是了解现有机制的重要一步， 未来的治疗药物对多种病毒性疾病。