Understanding the Assembly of the Small Ribosomal Subunit in Bacteria

了解细菌中小核糖体亚基的组装

• 批准号: 288327-2012
• 负责人: Ortega, Joaquin
• 金额: $ 2.48万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=506042
• 关键词: Understanding; Assembly; Small; Ribosomal; Subunit

The ribosome catalyzes protein synthesis in bacterial cells. Proteins are the building blocks for cellular structures and also for the enzymes that perform the biochemical reactions necessary to sustain life. Considering our dependency from bacteria for production of food and biofuels and for manufacturing a large number of basic products for Canadians (antibiotics, vaccines, enzymes and chemicals for industry, etc), it is essential that key processes of the bacterial physiology, including the assembly of the ribosome, are well understood. Made of two subunits (called 30S and 50S) and containing more than 50 individual components, the ribosome constitutes one of the most intricate macromolecular machines found in living organisms. Previous studies from others and us have identified a number of protein factors that act at the late stages of assembly of the 30S subunit, making its maturation an extremely efficient process. However, their mechanisms of action have not been revealed. Our research program is aimed at establishing a functional model for four critical assembly factors, YjeQ, RbfA, RimM and Era. Earlier studies suggested that these factors act in concert during the late stages of maturation of the 30S subunit and that they constitute the main group of proteins boosting the efficiency of the assembly process. To this end, we are obtaining the cryo-electron microscopy structure of ribosome intermediates accumulated by single-gene deletion strains lacking these factors and will study their degree of structural similarity. Collectively, these structures will establish whether YjeQ, RbfA, RimM and Era act in the same or different stages of assembly. In addition, we will establish the specific order in which they bind to the 30S ribosomal subunit and the conformational changes that they induce. As our program progresses, we will study how additional assembly factors also assist the process of assembly and tune the function of YjeQ, RbfA, RimM and Era. Ultimately, the outcome of our research will delineate the sequence of events that made ribosome assembly a highly efficient process. Consequently, it will constitute a significant step forward in our understanding of one of the essential processes for life.

核糖体催化细菌细胞中蛋白质的合成。蛋白质是细胞结构的基石，也是执行维持生命所必需的生化反应的酶的基石。考虑到我们生产食品和生物燃料以及为加拿大人制造大量基本产品(抗生素、疫苗、酶和工业化学品等)对细菌的依赖，很有必要对细菌生理学的关键过程，包括核糖体的组装，有很好的了解。核糖体由两个亚基(称为30S和50S)组成，包含50多个单独的成分，构成了在生物体内发现的最复杂的大分子机器之一。其他人和我们之前的研究已经确定了一些蛋白质因子，它们在30s亚单位组装的后期阶段发挥作用，使其成熟成为一个极其有效的过程。然而，他们的作用机制还没有被揭示。我们的研究计划旨在建立四个关键组装因子YjeQ、RbfA、Rimm和Era的功能模型。早期的研究表明，这些因素在30S亚基成熟的后期协同作用，它们构成了主要的蛋白质组，提高了组装过程的效率。为此，我们正在获得缺乏这些因子的单基因缺失菌株积累的核糖体中间产物的冷冻电子显微镜结构，并将研究它们的结构相似程度。总的来说，这些结构将确定YjeQ、RbfA、Rimm和Era在组装的相同或不同阶段发挥作用。此外，我们还将确定它们与30S核糖体亚基结合的具体顺序以及它们引起的构象变化。随着我们计划的进展，我们将研究其他组装因子如何也有助于组装过程，并调整YjeQ、RbfA、Rimm和Era的功能。最终，我们的研究结果将描绘出使核糖体组装成为一个高效过程的一系列事件。因此，这将是我们在理解生命的基本过程之一方面向前迈出的重要一步。