Editosome Protein-RNA Interactions in Trypanosomes

锥虫中编辑体蛋白-RNA 相互作用

• 批准号: 7348341
• 负责人: Salvador Z. Tarun
• 金额: $ 5.2万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7348341
• 关键词: Animal Diseases; Binding; Biochemical; Biological; Complex; Facility Construction Funding Category; Gene Expression; Genes; Goals; Human; In Vitro; Mitochondrial RNA; Physiological; Property; Protein Binding; Proteins; RNA; RNA Binding; RNA Editing; RNA Interference; RNA Processing; RNA-Protein Interaction; Recombinant Proteins; Research; Role; Transgenic Organisms; Trypanosoma; Trypanosoma brucei brucei; base; cell growth; in vivo; mutant; protein folding

Trypanosoma brucei is a group of pathogenic protozoans causing a spectrum of infectious human and related animal diseases that can be fatal and economically devastating. The long-term goal is to elucidate the biological importance and biochemical function of the editosomal proteins KREPA6 and KREPA5 in mitochondrial RNA editing, a unique RNA processing mechanism in trypanosomes that is essential for their survival. The proposal's specific aims are (1) to determine the physiological effects of disrupting the expression of these genes, (2) assess their RNA binding properties, and (3) identify KREPA6 protein binding partners. Regulatable RNAi, gene knock-outs, and construction of mutant transgenic strains will be used to determine their effects on cell growth and survival, editosome integrity and RNA editing in vivo. Recombinant proteins will be used to characterize their RNA binding substrates and protein binding partners in vitro. These studies will help reveal how KREPA5 and KREPA6 might contribute to the coordination of RNA substrate binding activities and catalytic steps within the editosome during RNA editing, helping advance our mechanistic understanding of RNA editing in trypanosomes.

布氏锥虫是一组致病性原生动物，可引起一系列人类和人类感染 相关的动物疾病可能是致命的并具有经济破坏性。长期目标是阐明 编辑体蛋白 KREPA6 和 KREPA5 的生物学重要性和生化功能 线粒体 RNA 编辑是锥虫中独特的 RNA 加工机制，对于锥虫的生存至关重要 生存。该提案的具体目标是（1）确定破坏 这些基因的表达，(2) 评估它们的 RNA 结合特性，(3) 识别 KREPA6 蛋白结合 合作伙伴。可调控的 RNAi、基因敲除和突变转基因菌株的构建将用于 确定它们对细胞生长和存活、编辑体完整性和体内 RNA 编辑的影响。重组 蛋白质将用于在体外表征其 RNA 结合底物和蛋白质结合伴侣。 这些研究将有助于揭示 KREPA5 和 KREPA6 如何促进 RNA 的协调 RNA 编辑过程中编辑体内的底物结合活性和催化步骤，有助于推进我们的研究 对锥虫 RNA 编辑机制的理解。