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(申请人提供)：本实验室以前的工作集中在两个锥虫特异性RNA结合蛋白P34/P37在基本保守的核糖体生物发生途径中的作用。这里提出的工作将扩大我们对锥虫核糖体生物发生的独特特征这一更大问题的研究。我们的合作者，哥伦比亚大学的Frank博士最近的冷冻-EM研究表明，布鲁氏锥虫核糖体具有许多独特的特征，包括核糖体RNA中的大扩展片段，这些片段可能是锥虫特异性蛋白的对接部位，或者是原本保守的核糖体蛋白的锥体特异性延伸。然而，他们的研究是有限的，因为他们专注于细胞质核糖体，并使用相对苛刻的条件来制备它们。因此，尽管发现了核糖体“核心”蛋白的一些有趣的特征，但他们无法识别任何独特的核糖体蛋白。许多实验室已经确定了锥虫体内核糖体生物发生所必需的非“核心”RNA结合蛋白的特征。这表明，这些锥虫特异性蛋白(包括那些已识别的和那些尚未发现的)更多是外围的，或者有更短暂的联系，但对核糖体成熟是必不可少的。这里描述的研究将识别在生物发生过程中与核糖体相互作用的锥虫特定蛋白，特别是与这些独特的扩展片段相互作用。我们还将研究Exportin 1(XpoI)出口复合体的独特功能，该复合体负责从细胞核到细胞质的60s(可能是40s)出口，并确定是否也使用其他出口受体系统。我们的假设是，这些蛋白质的功能将为干预布氏毛滴虫核糖体组装提供有效的靶点。该项目的具体目标涉及以下问题。1.哪些独特的蛋白质与核糖体相互作用？2.哪些蛋白质与扩张段相互作用，它们是锥体特有的还是保守的？3.60S和40S复合体如何与锥体中的核输出复合体相互作用？锥虫引起的疾病的治疗受到缺乏新药以及耐药性、毒性以及现有少数几种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疗干预的新靶点。