Dissecting the Mechanisms of Regulation and Quality Control in Ribosome Assembly and the Consequences of their Failure

剖析核糖体组装的调控和质量控制机制及其失败的后果

• 批准号: 10640194
• 负责人: Katrin Karbstein
• 金额: $ 95.21万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640194
• 关键词: Address; Affect; Biochemical; Biogenesis; Cells; Cellular biology; Degradation Pathway; Disease; Ensure; Failure; Genetic; Genomics; Goals; Head; Human; Link; Mediating; Messenger RNA; Molecular Biology; Pathway interactions; Protein Biosynthesis; Proteins; Quality Control; RNA Folding; Regulation; Research; Ribosomal Proteins; Ribosomes; Role; Structure; System; Translating; Work; Yeasts; cancer cell; conformational conversion; disease phenotype; frontier; human disease; insight; model organism; novel; proteostasis; tool

PROJECT SUMMARY Ribosomes carry out protein synthesis in all cells, interpreting the information contained in the mRNA to produce the proper amount of the correct protein. In addition, ribosomes also mediate mRNA quality control. Thus, misassembled ribosomes can affect the sequence and abundance of proteins and mRNAs, thereby disrupting protein homeostasis. This can lead to a number of diseases, demonstrating the importance of ensuring ribosomes are accurately assembled, and produced in the correct numbers. Using a combination of biochemical, genetic, genomic and structural tools, we will (i) investigate mechanisms that promote proper incorporation of ribosomal proteins and folding of the RNA, (ii) dissect quality control pathways to identify and ultimately degrade misassembled intermediates and (iii) study how misassembled ribosomes promote disease. In the first part, we will build on our existing work and study the late assembly of the ribosomal head, as well as combine insights from recent structures and our biochemical work to understand how DEAD-box proteins are used to construct ATP-dependent regulatory switches to control major conformational transitions in early 40S biogenesis. In the second part, we will extend our work on quality control to investigate a possible proofreading mechanism and identify degradation pathways for misassembled intermediates, a novel frontier for the field. In the last part, we will investigate how ribosomes containing substoichiometric levels of two ribosomal proteins, Asc1 and Rps10, which are produced in cancer cells that lack sufficient amounts of the assembly factor Ltv1, promote disease. This work builds on a genetic system we have developed to separate ribosomes of distinct composition, and also takes into consideration the known roles of these proteins in mRNA quality control. In addition, we will also investigate the effects from dysregulation of ribosome numbers in disease. Together, the proposed work will link mechanistic insights into a fundamental problem of cell and molecular biology – how ribosomes are assembled, to human disease.

项目摘要 核糖体在所有细胞中进行蛋白质合成，将mRNA中包含的信息解释为 产生适当量的正确蛋白质。此外，核糖体还介导mRNA质量控制。 这是填料的核糖体会影响蛋白质和mRNA的序列和抽象，从而 破坏蛋白质稳态。这可能导致多种疾病，证明了 确保准确组装核糖体，并以正确的数字产生。 使用生化，遗传，基因组和结构工具的组合，我们将（i）研究机制 促进核糖体蛋白的正确掺入和RNA的折叠，（ii）剖析质量控制 识别并最终降低无机组装中间体的途径，（iii）研究了如何填充 核糖体促进疾病。 在第一部分中，我们还将以现有工作为基础，并研究核糖体头的晚会 正如最近结构的结合见解和我们的生化工作，以了解死盒蛋白如何 用于构建依赖ATP的调节开关，以控制早期的主要构象转变 40S生物发生。 在第二部分中，我们将扩展我们在质量控制方面的工作，以调查可能的校对 机理并确定了MIDSASSMBLED INTERMEDIATES的降解途径，该中间体是该领域的新型边界。 在最后一部分中，我们将研究如何含有两个核糖体的化学计量水平的核糖体 蛋白质，ASC1和RPS10，它们是在缺乏足够量的癌细胞中产生的 因子LTV1，促进疾病。这项工作建立在我们开发的遗传系统上，以分离核糖体 独特的组成，还考虑了这些蛋白质在mRNA质量中的已知作用 控制。此外，我们还将研究核糖体数量失调中疾病中的影响。 拟议的工作一起将机械洞察力与细胞和分子的基本问题联系起来 生物学 - 核糖体如何组装成人类疾病。