Targeting the trypanosomal preribosomal complex

靶向锥虫前核糖体复合物

• 批准号: 9280621
• 负责人: Noreen Williams
• 金额: $ 28.62万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9280621
• 关键词: Affect; Affinity Chromatography; African Trypanosomiasis; Bacterial Infections; Binding; Biogenesis; Biological Assay; Cell Line; Cell Nucleolus; Cells; Chagas Disease; Characteristics; Collaborations; Complex; Computer Simulation; Defect; Development; Disease; Drug Targeting; Fluorescence Resonance Energy Transfer; Genetic Transcription; Goals; Homologous Gene; In Vitro; Knowledge; Laboratories; Leishmania; Leishmaniasis; Molecular; Mutation; Nucleoplasm; Organism; Parasites; Pharmacotherapy; Protein Biosynthesis; Proteins; RNA; RNA Interference; RNA Polymerase I; RNA Polymerase III; RNA-Protein Interaction; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Site; Structure; System; Trypanosoma; Trypanosoma brucei brucei; Trypanosoma cruzi; Work; drug development; high throughput screening; in vivo; insight; member; novel; particle; pathogen; protein function; protein protein interaction; public health relevance; rRNA Precursor; scaffold; screening

 DESCRIPTION (provided by applicant): Ribosomes are comprised of both rRNAs, which perform the catalytic function of protein synthesis, and a large number of proteins that form the structural scaffold. Transcription of the 35S rRNA precursor of 28S, 18S, and 5.8S by RNA polymerase I take place in the nucleolus where ribosomal assembly occurs. Only 5S rRNA is usually transcribed in the nucleoplasm by RNA polymerase III and brought into the nucleolus together with ribosomal protein L5. In the only well characterized eukaryotic system, Saccharomyes cerevisiae, two assembly factors, Rpf2 and Rrs1 as well as ribosomal protein, L11, are required to bring the early preribosomal (5S rRNA and L5) to the assembling ribosome. Our laboratory has defined the earliest preribosomal particle in trypanosomes which contains the conserved components 5S rRNA and L5 as well as the trypanosome-specific proteins, P34/P37. We have identified key determinants of this novel tri-molecular complex as well as unique features of the complex that we hypothesize are essential to its structure and function. We have also shown that both L5 and P34/P37 are essential to the formation of functional ribosomes and therefore, the viability of T. brucei. In this proposal we will identify and characterize the assembly components of the preribosomal particle and use our knowledge to define potential targets for chemotherapeutic approaches. Our hypothesis is that the interactions between the components of the essential preribosomal complex will provide valid targets for chemotherapeutic disruption of ribosomal assembly in T. brucei. The specific aims of the project are to: 1. Determine the critical proteins that enable the association of the 5S rRNA-containing complex with the ribosome and their function in ribosome biogenesis. 2. Define the interaction network among the pre-ribosomal complex components and examine how the disruption affects the function of the complex within the cell. 3. Develop FRET approaches to study RNA-protein interaction, in vivo protein-protein interactions, and screen for small interfering molecules. Although the ribosome is highly conserved, subtle differences between the host and pathogen have enabled the development of drugs specifically targeting pathogen ribosome assembly. Many existing drugs for the treatment of bacterial infections specifically bind to the functionally relevant sites of the bacterial ribosome. The pre-ribosomal complex that is the focus of this proposal is essential to trypanosome ribosome assembly and function and to the survival of the parasite. Proteins within the complex are either unique to trypanosomes (P34/P37) or contain features clearly distinct from those of the host (L5). Our work will also continue to provide new insights that will impact the broader field of eukaryotic ribosomal biogenesis. Moreover, the unique features of the complex will allow us to define and exploit this target for chemotherapeutic development.

 描述（由适用提供）：核糖体均累积了rRNA的核糖体，它们执行蛋白质合成的催化功能，以及形成结构支架的大量蛋白质。 RNA聚合酶I的35S rRNA前体的转录在发生核糖体组装的核心素中。 RNA聚合酶III通常仅在核等离子膜中转录5s rRNA，并与核糖体蛋白L5一起进入核olus。在唯一具有良好特征的真核系统中，需要酿酒酵母，两个组装因子RPF2和RRS1以及核糖体蛋白L11，以将早期的前蛋白体（5S rRNA和L5）带到组装核糖体中。我们的实验室定义了锥虫中最早的前粒子颗粒，其中包含保守的成分5S rRNA和L5以及锥虫特异性蛋白p34/p37。我们已经确定了这种新型的三分子复合物以及复合物的独特特征的关键决定者，我们假设对其结构和功能至关重要。我们还表明，L5和p34/p37均对形成功能性核糖体至关重要，因此，T。Brucei的生存能力。在此提案中，我们将确定并表征前粒子的组件成分，并利用我们的知识来定义化学治疗方法的潜在靶标。我们的假设是，基本前体复合物的组成部分之间的相互作用将为核糖体组装的化学治疗破坏提供有效的靶标。该项目的具体目的是：1。确定能够使5S rRNA复合物与核糖体及其在核糖体生物发生中的功能的关键蛋白。 2。定义核糖体复合物成分之间的相互作用网络，并检查破坏如何影响复合物在细胞内的功能。 3。开发研究RNA蛋白相互作用，体内蛋白 - 蛋白质相互作用以及筛选小的干扰分子的方法。尽管核糖体是高度保守的，但宿主和病原体之间的细微差异已使专门针对病原体核糖体组装的药物的发展。许多现有的用于治疗细菌感染的药物特异性结合了细菌核糖体功能相关部位。这是该建议的重点的核糖体复合物对于锥虫核糖体的组装和功能以及寄生虫的存活至关重要。复合物中的蛋白质要么是锥虫（p34/p37）独有的，要么包含与宿主（L5）明显不同的特征。我们的工作还将继续提供新的见解，从而影响真核核糖体生物发生的更广泛领域。此外，该综合体的独特特征将使我们能够定义和利用这一目标进行化学治疗发展。