Mechanisms of Nascent Polypeptide-Mediated Translational Regulation

新生多肽介导的翻译调控机制

• 批准号: 8393462
• 负责人: Mee-Ngan F Yap
• 金额: $ 23.41万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-05 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8393462
• 关键词: Biochemical; Biochemical Reaction; Biological; Co-Immunoprecipitations; Escherichia coli; Eukaryota; Gene Expression; Goals; Hereditary Disease; Human Genetics; Lead; Left; Life; Mediating; Messenger RNA; Methods; Modeling; Molecular; Movement; Mutagenesis; Mutation; Open Reading Frames; Organism; Peptides; Phase; Play; Process; Prokaryotic Cells; Protein Biosynthesis; Proteins; Proteome; Proteomics; Quality Control; Ribosomal Proteins; Ribosomes; Role; Signal Transduction; Staging; Stress; System; Techniques; Time; Transcript; Translational Regulation; Translations; Work; attenuation; bacterial genetics; comparative; crosslink; genome-wide; human disease; insight; interest; novel; polypeptide; protein aminoacid sequence; response

Ribosome movement on its template transcript plays an important role in mRNA quality control and translational regulation. Surprisingly, some regulatory nascent polypeptides stall their own translation when they are still inside the ribosome tunnel, a path that every newly synthesized protein traverses to leave the large ribosomal subunit. Despite the universality and structural conservation of the tunnel, its function is still rather mysterious. It is unclear how the tunnel deciphers the arrest signals and discriminates non-regulatory peptides from regulatory peptides. During the K99 phase I used E. coli SecM as a model to investigate how the arrest sequence motif is recognized inside the tunnel and how the timing of the release of arrest is achieved. In the ROO phase, we aim to advance our understanding of translatlonal attenuation by investigating the biological activities of the ribosome tunnel by characterizing the regulatory role of small peptides and by identifying peptide sequences within larger proteins that promote ribosome stalling/pausing. Specifically we will use an Integrative approach that combines comparative proteomics, bacterial genetics, biochemical methods and genome-wide ribosome profiling to elucidate the fundamental principles of these processes. This work will not only provide new information on the molecular responses of the tunnel to disparate nascent chains, but will also offer a broader view of the diversity of translational control systems and the translation machinery that is conserved in all living systems.

核糖体在其模板转录本上的运动在 mRNA 质量控制和 翻译监管。令人惊讶的是，一些调节性新生多肽在 它们仍然位于核糖体隧道内，这是每个新合成的蛋白质离开核糖体时都要经过的路径。 大核糖体亚基。尽管隧道具有普遍性和结构保护性，但其功能仍然存在 相当神秘。目前尚不清楚隧道如何破译逮捕信号并区分非监管人员 来自调节肽的肽。在 K99 阶段，我使用大肠杆菌 SecM 作为模型来研究如何 逮捕序列基序在隧道内被识别，以及释放逮捕的时间是如何的 实现了。在 ROO 阶段，我们的目标是通过以下方式增进我们对平移衰减的理解： 通过表征小分子的调节作用来研究核糖体通道的生物活性 肽并通过识别较大蛋白质内促进核糖体停滞/暂停的肽序列。 具体来说，我们将使用一种综合方法，结合比较蛋白质组学、细菌遗传学、 生化方法和全基因组核糖体分析来阐明这些的基本原理 流程。这项工作不仅将提供有关隧道分子响应的新信息 不同的新生链，但也将为翻译控制系统的多样性提供更广阔的视野 以及所有生命系统中保存的翻译机制。