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）：冠状病毒引起鸟类和动物的呼吸道和胃肠道疾病。这些RNA病毒在遗传上是可变的，并且可以迅速发展以感染人类，有时会导致严重的急性呼吸道疾病。冠状病毒是通过涉及细胞内病毒形成以及向外部环境的新型运输路线进行的策略来分开的。对这些晚期感染事件的进一步了解将确定治疗干预的策略。我们已经发现，在组装和 /或分泌阶段抑制了小鼠肝炎冠状病毒，蛋白酶体抑制剂的无毒浓度非常低。我们假设抑制性机制涉及细胞的泛素耗竭，这是蛋白酶体抑制剂的已知作用，因为我们发现病毒分泌的核心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冠状病毒是否对蛋白酶体抑制剂过敏，然后解决E蛋白的泛素结合是否对冠状病毒形态发生还是从细胞中驱动的核心。我们的实验将专门评估抑制剂和泛素作用的源头是否处于病毒分泌阶段，并将解决泛素修饰的新假设，即泛素修饰将病毒填充细胞器的运输转移到释放病毒货物的细胞表面上。我们的发现将揭示囊泡形成和细胞器转运的新型细胞生物学特征，还将告知我们在晚期感染阶段挫败冠状病毒的潜力。公共卫生相关性：冠状病毒引起人类和家养动物的呼吸道疾病，在严重的急性呼吸综合症冠状病毒的情况下，症状可能危及生命。冠状病毒以新颖的方式形成，通过组装成细胞器囊泡，然后在囊泡融合到质膜时从细胞中排出。关于这一过程的发生知之甚少，该提案的目的是从细胞中揭示冠状病毒组装和分泌的机制，从而发现了以这种基本感染阶段进行干预的靶标。