Role of Coronavirus Membrane Protein Carboxy Tail Virus Assembly

冠状病毒膜蛋白羧基尾病毒组装的作用

• 批准号: 7555074
• 负责人: Ariel L. Arndt
• 金额: $ 3.62万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-20 至 2010-12-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7555074
• 关键词: Acute Pneumonia; Affect; Amino Acids; Animals; Antiviral Agents; Asorbicap; Automobile Driving; Biochemical; C-terminal; Cattle; Charge; Chiroptera; Common Cold; Confocal Microscopy; Coronaviridae; Coronavirus; Country; Cytoplasm; Cytoplasmic Tail; Development; Diagnostic; Domestic Fowls; E protein; Economics; Endoplasmic Reticulum; Enteral; Epidemic; Family; Family member; Family suidae; Genome; Genomics; Genotype; Goals; Golgi Apparatus; Growth; Health; Healthcare; Human; Individual; Industry; Infection; Kinetics; Knowledge; Length; Mediating; Membrane; Membrane Proteins; Morphology; Murine hepatitis virus; N-terminal; Nucleocapsid; Pathogenesis; Phenotype; Phosphoproteins; Play; Prevention; Process; Protein Binding; Proteins; RNA; RNA Viruses; Reagent; Research; Role; Scientist; Severe Acute Respiratory Syndrome; Structural Protein; Structure; Surface; Tail; Transmembrane Domain; Vaccines; Viral; Viral Matrix Proteins; Viral Proteins; Virion; Virus; Virus Assembly; Virus-like particle; economic impact; env Gene Products; human coronavirus; insight; interest; mortality; multiple myeloma M Protein; mutant; protein protein interaction; protein structure; respiratory; social; vaccine development

DESCRIPTION (provided by applicant): Coronaviruses are a medically significant group of enveloped, positive stranded RNA viruses that cause primarily respiratory and enteric infections in humans and many animals. The overall focus of this proposal is to understand the mechanism and function of coronavirus proteins in virus assembly. The virion envelope contains at least two major structural proteins, the membrane (M) and spike (S) proteins. Additionally, a few molecules of the envelope (E) protein are also present in the envelope. The nucleocapsid (N) phosphoprotein encapsidates the genomic RNA packaged inside the envelope. The N protein binds the genomic RNA to form a helical nucleocapsid. The most abundant structural protein in the virion envelope is the M protein. It interacts with itself, other structural proteins and the nucleocapsid and therefore plays a key role in viral assembly. The M protein contains a short N-terminal domain outside the virion, three transmembrane domains and a long C-terminus domain inside the virion. The C-terminus contains a long amphipathic domain and a highly charged hydrophilic tail. The amphipathic domain has been implicated to play a role in various viral protein-protein interactions, including M-S and M-E associations. Additionally, the extreme C-terminal -25 amino acids of M have been shown to be functionally important in assembly through interactions with the nucleocapsid. The specific residues and requirements that are involved in mediating these protein-protein interactions are not fully defined and understood. The specific aims of the proposed research are to determine the role of a conserved domain and charged residues within the M protein cytoplasmic tail that are important in coronavirus assembly. To achieve this goal, a panel of mutant viruses will be generated with various changes in charged residues and the conserved domain within the M tail. All of the mutants will be analyzed in the context of the full-length mouse hepatitis virus (MHV-CoV)-A59 infectious clone. Mutants will be studied for plaque morphology and growth kinetics compared to wild-type virus. Biochemical analysis, coimmunoprecipitation (co-IP) and confocal microscopy will be used to elucidate the mechanistic roles of M in assembly, in terms of specific residue and structure requirements. The proposed study will provide information which is applicable to identifying potential antiviral targets. Antiviral development against coronaviruses, including the recently emerged severe acute respiratory syndrome coronavirus (SARS-CoV), will provide a significant improvement in health care treatment options. Antiviral reagents and vaccines directed towards coronaviruses will aid in the treatment and prevention of respiratory and enteric infections, thus contributing to global improvement of human health.

描述（由申请人提供）：冠状病毒是一组具有医学意义的阳性，滞留的RNA病毒，主要引起人类和许多动物的呼吸和肠道感染。该建议的总体重点是了解病毒组装中冠状病毒蛋白的机制和功能。病毒座包膜至少包含两个主要的结构蛋白，即膜（M）和尖峰（S）蛋白。此外，包膜中也存在一些包膜（E）蛋白的分子。 Nucleocapsid（N）磷蛋白封装了包装在包膜内的基因组RNA。 N蛋白结合基因组RNA形成螺旋核蛋白酶。病毒座包膜中最丰富的结构蛋白是M蛋白。它与自身相互作用，其他结构蛋白和核素蛋白质，因此在病毒组装中起关键作用。 M蛋白在病毒座之外包含一个短的N末端结构域，三个跨膜结构域和病毒体内的一个长C末端结构域。 C末端包含一个长的两亲域和高电荷的亲水性尾巴。两亲域已被牵涉到在包括M-S和M-E关联在内的各种病毒蛋白 - 蛋白质相互作用中发挥作用。另外，通过与核蛋白酶的相互作用，M的极端C末端-25氨基酸在组装中在功能上很重要。介导这些蛋白质蛋白质相互作用的特定残基和要求尚未完全定义和理解。拟议的研究的具体目的是确定在冠状病毒组装中重要的M蛋白质细胞质尾巴中保守域和带电残基的作用。为了实现这一目标，将产生一系列突变病毒，并在电荷残基和M尾部内的保守域进行各种变化。所有突变体将在全长小鼠肝炎病毒（MHV-COV）-A59传染性克隆的背景下进行分析。与野生型病毒相比，将研究突变体的斑块形态和生长动力学。生化分析，共免疫沉淀（CO-IP）和共聚焦显微镜将根据特定的残基和结构要求来阐明M组装中M的机械作用。拟议的研究将提供适用于识别潜在抗病毒靶标的信息。针对冠状病毒的抗病毒发育，包括最近出现的严重急性呼吸道综合征冠状病毒（SARS-COV），将在医疗保健治疗方案方面显着改善。针对冠状病毒的抗病毒试剂和疫苗将有助于治疗和预防呼吸道和肠道感染，从而有助于全球人类健康的改善。