UBIQUITIN AND CELLULAR FACTORS IN CORONAVIRUS ASSEMBLY

冠状病毒组装中的泛素和细胞因子

基本信息

批准号：
7860419
负责人：
Thomas Miller Gallagher
金额：
$ 18.69万
依托单位：
LOYOLA UNIVERSITY CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-05 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7860419
关键词：
Acute Address Animals Antineoplastic Agents Applications Grants Biological Birds Bortezomib Cell membrane Cell secretion Cell surface Cells Complex Containment Coronaviridae Coronavirus Coronavirus Infections Cytoplasmic Tail Defect Disease Domestic Animals E protein Environment Event Exhibits Experimental Designs FDA approved Family Future Gastrointestinal Diseases Hepatitis Human Infection Intervention Life Lung diseases Lysine MG132 Mediating Modeling Modification Molecular Genetics Morphogenesis Mus Organelles Process Proteasome Inhibitor Proteins RNA Viruses Recombinants Research Route Severe Acute Respiratory Syndrome Staging Symptoms Therapeutic Intervention Ubiquitin Ubiquitination Vesicle Viral Viral Proteins Virion Virus Virus Assembly base clinically significant cofactor human coronavirus inhibitor/antagonist member novel particle programs prototype public health relevance research study respiratory trafficking ubiquitin ligase

项目摘要

DESCRIPTION (provided by applicant): Coronaviruses cause respiratory and gastrointestinal diseases in birds and animals. These RNA viruses are genetically variable and can rapidly evolve to infect humans, sometimes causing severe acute respiratory disease. The coronaviruses are set apart in their assembly and secretion from cells by a strategy involving intracellular virus formation and perhaps novel trafficking routes to the outside environment. Further understanding of these late infection events will identify strategies for therapeutic intervention. We have discovered that a prototype mouse hepatitis coronavirus is inhibited at the assembly and / or secretion stages by very low nontoxic concentrations of a proteasome inhibitor. We hypothesize that the inhibitory mechanism involves ubiquitin depletion from cells, a known effect of proteasome inhibitors, because we discovered that the viral E proteins that are central to virus secretion are ubiquitinated on two lysine residues. Our aims are to determine whether pathogenic human coronaviruses are similarly hypersensitive to proteasome inhibitors and then address whether the ubiquitin conjugation of E proteins is central to coronavirus morphogenesis or expulsion out of cells. Our experiments will specifically evaluate whether the locus of inhibitor and ubiquitin action are at the virus secretion stages and will address the novel hypothesis that ubiquitin modifications direct the trafficking of virus-filled organelles to cell surfaces where the virus cargo is liberated. Our findings will reveal novel cell biological features of vesicle formation and organelle transport and will also inform us about the potential to thwart coronaviruses at late infection stages. PUBLIC HEALTH RELEVANCE: Coronaviruses cause respiratory diseases in humans and domesticated animals and in cases of severe acute respiratory syndrome coronavirus the symptoms can be life threatening. Coronaviruses form in a novel fashion by assembling into organellar vesicles and then expelling from cells when the vesicles fuse to the plasma membrane. As little is known about how this process occurs, the proposal aims to unravel mechanisms of coronavirus assembly and secretion from cells and thereby uncover targets for intervention with this essential infection stage.

描述（由申请人提供）：冠状病毒会引起鸟类和动物的呼吸道和胃肠道疾病。这些 RNA 病毒具有遗传变异，可以迅速进化以感染人类，有时会导致严重的急性呼吸道疾病。冠状病毒在细胞的组装和分泌过程中是通过一种涉及细胞内病毒形成以及可能新颖的运输到外部环境的途径的策略来区分的。对这些晚期感染事件的进一步了解将确定治疗干预策略。我们发现，原型小鼠肝炎冠状病毒在组装和/或分泌阶段被极低无毒浓度的蛋白酶体抑制剂抑制。我们假设抑制机制涉及细胞中泛素的消耗，这是蛋白酶体抑制剂的已知作用，因为我们发现对病毒分泌至关重要的病毒 E 蛋白在两个赖氨酸残基上被泛素化。我们的目的是确定致病性人类冠状病毒是否同样对蛋白酶体抑制剂过敏，然后确定 E 蛋白的泛素缀合是否是冠状病毒形态发生或从细胞中排出的核心。我们的实验将专门评估抑制剂和泛素作用位点是否处于病毒分泌阶段，并将解决新的假设，即泛素修饰引导充满病毒的细胞器运输到释放病毒货物的细胞表面。我们的研究结果将揭示囊泡形成和细胞器运输的新细胞生物学特征，还将告诉我们在感染后期阻止冠状病毒的潜力。公共卫生相关性：冠状病毒会引起人类和家养动物的呼吸道疾病，在严重急性呼吸综合征冠状病毒的情况下，症状可能会危及生命。冠状病毒以一种新颖的方式形成，通过组装成细胞器囊泡，然后当囊泡与质膜融合时从细胞中排出。由于对这一过程如何发生知之甚少，该提案旨在揭示冠状病毒组装和细胞分泌的机制，从而发现干预这一重要感染阶段的目标。