Cardioviral Proteases and Comparative Genome Structure

心脏病毒蛋白酶和比较基因组结构

• 批准号: 8197069
• 负责人: ANN C. PALMENBERG
• 金额: $ 35.63万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8197069
• 关键词: Amaze; Antiviral Agents; Back; Binding; Biochemistry; Biology; Brain; Cardiovirus; Cell Nucleolus; Cell Nucleus; Cells; Cessation of life; Complex; Cytoplasm; Disasters; Disease; Encephalomyocarditis virus; Enzyme Precursors; Enzymes; Event; Evolution; Family; Family Picornaviridae; Family member; FarGo; Fertility; Foundations; Genetic Transcription; Genetic Translation; Genome; Goals; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Heart; Homologous Gene; Host Defense; Hour; Immune response; Immune system; Individual; Infection; Intestines; Investigation; Laboratories; Lead; Life; Lytic; Messenger RNA; Molecular; Morphogenesis; Mus; Natural Immunity; Nuclear Pore; Nucleic Acids; Occupations; Outcome; Pancreas; Pathway interactions; Peptide Hydrolases; Phase; Phosphorylation; Phosphotransferases; Picornaviridae Infections; Polyproteins; Pore Proteins; Process; Proteins; Proteolytic Processing; RNA; Ribosomes; Running; Signal Transduction; Structural Protein; Structure; System; Translations; Viral; Viral Proteins; Virus; Virus Diseases; War; Work; analog; arm; cohort; comparative; experience; extracellular; falls; fighting; inhibitor/antagonist; intercellular communication; killings; member; novel; nucleocytoplasmic transport; prevent; programs; protein function; research study; spelling; trafficking

The goals of this investigation are to explore and define the relationship of the cardiovirus genus to other members of the picornavirus family and to exploit the unique features of cardioviruses to examine fundamental molecular questions about picornavirus translation, proteolytic processing, morphogenesis and host interaction. The RNA picornaviruses are one of the best understood and most thoroughly accessible experimental systems in all of biology. Natural infections with cardioviruses, like encephalomyocarditis virus (EMCV), kill nearly every cell in the brain, pancreas and heart of a mouse, within 3 days. The virus does this with apparent impunity to cellular antiviral defenses by subverting innate immunity traps and crippling the capacity of an infected cell to mount a defense or trigger an alarm. The molecular battleground inside infected cells pits viral protease 3Cpro and two unique cardiovirus proteins, L and 2A, enzymes honed by evolution for their special anti-cellular purposes, against the complete array of innate host defenses. The outcome rarely varies. Within 2-3 hours of infection EMCV brings to a halt cellular mRNA transcription, cap- dependent mRNA translation, antiviral signal transduction, and active protein/RNA exchange between the nucleus and cytoplasm. The virus replicates with fecundity and the cell dies before it ever triggers an alarm. At the ultimate molecular level, the activities of these proteins instigate the cascade of events that set off or prevent an episode of disease. The next phase of this project will examine the biochemistry and molecular pathways of EMCV L (Leader), the first viral (or cellular) protein known to bind and inactivate Ran GTPase cycling, the crucial, ubiquitous regulatory system for all protein and nucleic acid trafficking into and out of the nucleus. The project also examines the pathways by which the presence of L, or L-Ran complexes activate a specific cohort of cellular kinases, and redirects them towards the phosphorylation of nuclear pore proteins (Nups). The specific aims are: (1) To resolve the NMR structure of Mengo L (Leader) protein as it interacts with Ran GTPase. (2) To characterize the biochemistry of cardiovirus L:Ran interactions which inhibit RanGDP/GTP cycling in cell-free extracts. (3) To identify within cells, the host kinases activated by L, which contribute to the abrogation of nucleocytoplasmic trafficking steps. (4) To define the molecular interactions of L with other viral proteins (2A and 3CD), and define the replication advantages to cardioviruses, for encoding a unique and potent inhibitor of Ran and of cellular protein and mRNA trafficking. These objectives build directly upon experimental foundations developed during the preceding 27 years of the program.

本研究的目的是探索和确定心脏病毒属与其他病毒的关系。 小核糖核酸病毒家族的成员，并利用心脏病毒的独特功能来检查 关于小核糖核酸病毒翻译、蛋白水解加工、形态发生和 宿主相互作用RNA小核糖核酸病毒是最好理解和最彻底的访问之一 所有生物学的实验系统。心脏病毒的自然感染，如脑心肌炎病毒 （EMCV），在3天内杀死小鼠脑、胰腺和心脏中的几乎每一个细胞。病毒就是这么做的 通过破坏先天免疫陷阱和削弱免疫系统， 受感染细胞的防御能力或触发警报的能力。内部的分子战场 感染的细胞使病毒蛋白酶3Cpro和两种独特的心脏病毒蛋白L和2A形成凹坑， 进化的特殊抗细胞的目的，对先天宿主防御的完整阵列。的 结果很少变化。在感染后2-3小时内，EMCV使细胞mRNA转录停止，帽- 依赖的mRNA翻译，抗病毒信号转导，以及活性蛋白/RNA之间的交换。 细胞核和细胞质。病毒以繁殖力进行复制，细胞在触发警报之前就死亡了。 在最终的分子水平上，这些蛋白质的活动引发了一系列事件， 防止疾病的发作。这个项目的下一阶段将研究生物化学和分子生物学。 EMCV L（Leader）是已知的第一种结合和抑制Ran GT3的病毒（或细胞）蛋白， 循环，至关重要的，无处不在的调节系统，所有的蛋白质和核酸运输进出 原子核该项目还研究了L或L-Ran复合物的存在的途径 激活一组特定的细胞激酶，并将它们重定向到核孔的磷酸化， 蛋白质（NUPS）。具体目标是：（1）解析Mengo L（Leader）蛋白的NMR结构， 与Ran GTweek互动。(2)为了表征心脏病毒L：Ran相互作用的生物化学， 抑制无细胞提取物中RanGDP/GTP循环。(3)为了在细胞内鉴定由L激活的宿主激酶， 其有助于消除核质运输步骤。(4)来定义分子 L与其他病毒蛋白（2A和3CD）的相互作用，并定义了复制优势， 心脏病毒，用于编码Ran和细胞蛋白质和mRNA的独特且有效的抑制剂 贩卖人口这些目标直接建立在前一阶段开发的实验基础上。 27年的节目。