Ribosomal biogenesis in trypanosomes

锥虫中的核糖体生物发生

• 批准号: 8670854
• 负责人: Noreen Williams
• 金额: $ 29.38万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8670854
• 关键词: Address; Affinity; Affinity Chromatography; African Trypanosomiasis; Antibiotics; Biogenesis; Biological Assay; Cell Nucleus; Chagas Disease; Complex; Core Protein; Cytoplasm; Disease; Docking; Intervention; Laboratories; Leishmania; Leishmaniasis; Methods; Nuclear Export; Nuclear Pore Complex; Oligonucleotides; Organism; Parasites; Pathway interactions; Peripheral; Pharmaceutical Preparations; Play; Process; Protein Binding; Proteins; RNA Interference; RNA-Binding Proteins; Resistance; Ribosomal Biogenesis Pathway; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Site; Structure; System; Toxic effect; Trypanosoma; Trypanosoma brucei brucei; Trypanosoma cruzi; Universities; Work; Yeasts; cost; exportin 1 protein; interest; particle; protein function; public health relevance; receptor

DESCRIPTION (provided by applicant): Previous work from this laboratory has focused on the role of two trypanosome-specific RNA binding proteins, P34/P37, in the largely conserved ribosomal biogenesis pathway. The work proposed here will expand our studies into the larger question of the unique features of trypanosome ribosomal biogenesis. Recent cryo-EM studies from our collaborator, Dr. Frank (Columbia University), have shown that the T. brucei ribosome possesses a number of unique features including large expansion segments in the ribosomal RNAs that suggest sites for docking of trypanosome-specific proteins or trypanosome-specific extensions of the otherwise conserved ribosomal proteins. However, their studies were limited in that they focused on the cytoplasmic ribosome and used relatively harsh conditions to prepare them. So although some interesting features of the ribosome "core" proteins were found, they were unable to identify any unique ribosomal proteins. A number of laboratories have characterized non-"core" RNA binding proteins that are essential to ribosomal biogenesis in trypanosomes. This suggests that these trypanosome-specific proteins (both those already identified and those yet to be discovered) are more peripheral or have a more transient association and yet are essential to ribosomal maturation. The studies described here will identify trypanosome-specific proteins that interact with the ribosome during the biogenesis pathway and, in particular, interact with these unique expansion segments. We will also examine the unique features of the exportin 1 (XpoI) export complex responsible for 60S (and likely 40S) export from the nucleus to the cytoplasm and determine whether other export receptor systems are also used. Our hypothesis is that the function of these proteins will provide valid targets for intervention in the ribosomal assembly in T. brucei. The specific aims of the project address the following questions. 1. What unique proteins interact with the ribosome? 2. Which proteins interact with the expansion segments and are they conserved or unique to trypanosomes? 3. How do the 60S and 40S complexes interact with the nuclear export complex in trypanosomes? Treatment of diseases caused by trypanosomes is severely limited by the lack of new drugs as well as resistance, toxicity, and issues with cost and administration for the few drugs currently available. Ribosomal structure and function have been identified as targets for antibiotics in other systems. By determining additional unique features of kinetoplastid ribosomal structure and biogenesis we will identify new targets for chemotherapeutic intervention that are desperately needed for these parasites.

描述（由申请人提供）：该实验室的先前工作集中在两个锥体特异性RNA结合蛋白P34/p37在很大程度上保守的核糖体生物发生途径中的作用。这里提出的工作将使我们的研究扩大到锥虫核糖体生物发生的独特特征的更大问题。我们的合作者Frank（哥伦比亚大学）的最近的低温EM研究表明，T. brucei核糖体具有许多独特的功能，包括核糖体RNA中的大型扩展段，建议对接锥虫特异性蛋白质或锥虫特异性蛋白或其他固化的固醇蛋白质蛋白质蛋白质的网站。但是，他们的研究受到限制，因为他们专注于细胞质核糖体，并使用相对恶劣的条件为它们做好准备。因此，尽管发现了核糖体“核心”蛋白的一些有趣特征，但他们无法识别任何独特的核糖体蛋白。许多实验室表征了非“核” RNA结合蛋白，这些蛋白对于锥虫中的核糖体生物发生至关重要。这表明这些锥虫特异性蛋白（均已鉴定出的蛋白质和尚未发现的蛋白质）更外围或具有更短暂的关联，但对于核糖体成熟至关重要。此处描述的研究将确定在生物发生途径期间与核糖体相互作用的锥虫特异性蛋白，尤其是与这些独特的扩张片段相互作用。我们还将检查导出1（XPOI）的导出复合物的独特特征，该复合物负责60s（可能是40s）从细胞核出口到细胞质，并确定是否还使用了其他导出受体系统。我们的假设是，这些蛋白质的功能将提供有效的靶标，以干预布鲁氏菌中的核糖体组装。项目的具体目的解决了以下问题。 1。哪些独特的蛋白质与核糖体相互作用？ 2。哪些蛋白与膨胀片段相互作用，它们是保守的还是锥虫的独特？ 3。60s和40s的复合物如何与锥虫中的核出口复合物相互作用？锥虫引起的疾病的治疗受到缺乏新药，耐药性，毒性以及目前可用的少数药物的成本和管理问题的严重限制。核糖体的结构和功能已被确定为其他系统中抗生素的靶标。通过确定动力质体核糖体结构和生物发生的其他独特特征，我们将确定这些寄生虫迫切需要的化学治疗干预措施的新目标。