A novel preribosomal complex in trypanosomes

锥虫中的一种新型前核糖体复合物

• 批准号: 7853707
• 负责人: Noreen Williams
• 金额: $ 30.79万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7853707
• 关键词: African Trypanosomiasis; Antibiotics; Bacteria; Binding; Binding Proteins; Biogenesis; Biological Assay; Cell Nucleolus; Cell Nucleus; Chagas Disease; Chemicals; Complex; Computer Simulation; Data; Databases; Disease; Drug Delivery Systems; Energy Transfer; Fluorescence; Gel; Goals; Homologous Gene; Human; Immune; In Vitro; Intervention; Ketolides; Leishmania; Leishmaniasis; Macrolides; Maintenance; Maps; Mediating; Methods; Molecular; Mutation; Nucleoplasm; Organism; Parasites; Pathway interactions; Play; Process; Protein Binding Domain; Protein Fragment; Proteins; RNA-Binding Proteins; Research; Resolution; Ribosomal Biogenesis Pathway; Ribosomal RNA; Role; Route; Screening procedure; Signal Transduction; Small RNA; Staging; Structure; System; Technology; Testing; Transcription Factor TFIIIA; Trypanosoma; Trypanosoma brucei brucei; Trypanosoma cruzi; Work; base; chemotherapy; domain mapping; in vivo; migration; novel; particle; public health relevance; research study; ribosomal protein L5; trafficking

DESCRIPTION (provided by applicant): We have shown that two trypanosome-specific RNA binding proteins, P34 and P37, play an essential role in the ribosomal biogenesis pathway in T. brucei. This represents a novel finding since the ribosomal pathway is highly conserved. To exploit this observation for the ultimate goal of targeting the T. brucei ribosomal pathway in chemotherapy, we must understand the mechanism by which these proteins act in the pathway. In recent work, we have shown that P34 and P37 interact with both L5 and 5S rRNA, the highly conserved components of the preribosomal particle. Loss of P34/P37 leads to a decrease in 5S rRNA levels and other effects in ribosomal formation. The objective of this proposal is to determine the critical residues involved in this interaction and develop a method to identify molecules that disrupt this interaction. Our hypothesis is that the molecular interactions of these proteins will provide a valid target for intervention in ribosomal assembly in T. brucei. The specific aims of the project are to: 1. Determine the critical molecular determinants on L5 and P34/P37 that are required for participation in the T. brucei preribosomal complex. 2. Determine the critical regions on 5S rRNA that interact with L5 and P34/P37 in the T. brucei preribosomal complex. 3. Determine whether the interaction of P34/P37 with L5 and 5S rRNA is required for trafficking of the complex to the nucleolus. 4. Develop a FRET (fluorescence energy transfer) based screen for the disruption of the interaction between P34/P37, L5, and 5S rRNA in the preribosomal particle. Differences in ribosomal structure and function between bacteria and the infected human host have provided targets for macrolide and ketolide antibiotics. Since this preribosomal particle is unique to kinetoplastids and play an essential role in the ribosomal structure and biogenesis in these organisms, mechanistic studies with this complex will allow us to define a strategic target for chemical intervention. PUBLIC HEALTH RELEVANCE: The protozoan parasites, the Kinetoplastids, include the organisms Trypanosoma brucei, Trypanosoma cruzi and Leishmania, which cause serious diseases in much of the world (African sleeping sickness, Chagas' disease and leishmaniasis, respectively). This study will examine a unique preribosomal complex occurring in these parasites with the goal of developing an assay to identify drugs that target the parasite but not host complex.

描述（由申请人提供）：我们已经表明，两个锥体特异性RNA结合蛋白P34和P37在T. Brucei的核糖体生物发生途径中起着至关重要的作用。这代表了一个新颖的发现，因为核糖体途径高度保守。为了利用这一观察结果，以实现化学疗法中靶向布鲁氏菌核糖体途径的最终目标，我们必须了解这些蛋白在途径中起作用的机制。在最近的工作中，我们表明p34和p37与L5和5S rRNA相互作用，L5和5S rRNA是前粒子的高度保守成分。 p34/p37的丧失导致5S rRNA水平降低，核糖体形成其他影响。该提案的目的是确定这种相互作用中涉及的关键残基，并开发一种方法来识别破坏这种相互作用的分子。我们的假设是，这些蛋白质的分子相互作用将为干预核体组装的有效靶标。该项目的具体目的是：1。确定参与T. brucei t. brucei t. t. brucei preribosomal复合物所需的L5和p34/p37上的临界分子决定因素。 2。确定5S rRNA上与L5和p34/p37相互作用的临界区域。 3。确定是否需要将p34/p37与L5和5S rRNA的相互作用进行运输到核仁。 4。开发基于FRET（荧光能传递）的屏幕，以破坏前粒子中p34/p37，L5和5S rRNA之间的相互作用。细菌与感染人类宿主之间的核糖体结构和功能的差异为大环内酯类和酮酸抗生素提供了靶标。由于这种前粒子是动力质体特有的，并且在这些生物体中的核糖体结构和生物发生中起着至关重要的作用，因此使用该复合物的机械研究将使我们能够为化学干预的战略目标定义。 公共卫生相关性：原生动物寄生虫，动力质体，包括Brucei锥虫锥虫，Cruzi锥虫和利什曼尼亚锥虫，它们在世界上引起严重的疾病（分别为非洲睡眠疾病，Chagas的疾病和Leishmaniasis）。这项研究将检查在这些寄生虫中发生的独特的前体络合物，目的是开发一种测定法以识别靶向寄生虫但不是宿主复合物的药物。