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: 8664595
• 负责人: William T Jackson
• 金额: $ 35.96万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8664595
• 关键词: Appearance; Autophagocytosis; Autophagosome; Capsid Proteins; Cell membrane; Cells; Cellular Membrane; Cellular Stress; Characteristics; Complex; Cytosol; DNA Sequence Rearrangement; Data; Development; Encapsulated; Environment; Enzymes; Extracellular Space; Family; Family Picornaviridae; Future; Generations; Genome; Genomics; Goals; Golgi Apparatus; Guanine Nucleotide Exchange Factors; 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; Secretory Vesicles; Site; Staging; Testing; Therapeutic; Therapeutic Agents; Time; Translations; Transport Vesicles; Vesicle; Viral; Virion; Virus; Virus Diseases; Virus Replication; Work; base; biological adaptation to stress; human disease; innovation; insight; novel; pathogen; prevent; research study; therapeutic target; viral RNA

DESCRIPTION (provided by applicant): 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 but is dispensable for PV entry, translation, and RNA replication. We have found that 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 morphogenesis of COPII-like secretory vesicles used for RNA replication, and 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 identif 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.

描述（由申请人提供）：迄今为止研究的所有正链RNA病毒都重排细胞膜以促进自身复制。这些重排的一个原因是，这些病毒复制与细胞膜相关的基因组RNA。脊髓灰质炎病毒（PV）是医学上重要的正链RNA病毒宿主的一种模型，在PV中，已经确定了两种不同类型的囊泡。一类是单膜囊泡，类似于COPII分泌运输囊泡，与病毒RNA复制蛋白相关。第二种囊泡是双膜的，也与病毒RNA复制蛋白相关，类似于自噬体，自噬体是一种由细胞稳态和应激反应途径诱导的细胞器，称为自噬。自噬体样囊泡被病毒特异性诱导并促进PV的产生。该项目的长期目标是了解小核糖核酸病毒对细胞膜重排的机制和后果。此应用程序的目标是确定所扮演的角色