Mechanisms of Translation Control in Humans

人类翻译控制机制

• 批准号: 10552291
• 负责人: JAMIE H CATE
• 金额: $ 50.57万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-07 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552291
• 关键词: Affect; Area; Back; Behavior; Binding; Biochemical; Cells; Cellular immunotherapy; Elements; Engineering; Eukaryota; Eukaryotic Initiation Factor-3; Foundations; Future; Genotype; Glean; Human; Investigation; Life; Maps; Messenger RNA; Molecular; Pathway interactions; Peptide Initiation Factors; Phenotype; Process; Protein Biosynthesis; Proteins; Quality Control; RNA; Ribosomal Frameshifting; Ribosomes; Role; Speed; Structure; T cell regulation; T-Cell Activation; T-Cell Development; T-Cell Receptor; Therapeutic; Tissues; Trans-Activators; Translation Initiation; Translational Regulation; Translational Repression; Translations; design; drug discovery; drug-like compound; experimental study; human disease; improved; insight; interest; polypeptide; small molecule

ABSTRACT Protein synthesis, or translation, connects genotype to phenotype in all forms of life. The Cate lab has a longstanding interest in the mechanisms of protein synthesis, from universal principles gleaned from bacterial translation to the basis of translation regulation in humans. This application tackles fundamental questions about how translation is regulated in humans. We propose to explore the regulation of translation initiation in specific cells and tissues, and mechanisms of translation elongation that affect the speed and accuracy of the ribosome. We think these two broad lines of investigation will lead to many discoveries about protein synthesis that could eventually be leveraged to treat human disease. The canonical mechanism of translation initiation in eukaryotes involves many general translation initiation factors. We recently discovered that one of these–eukaryotic initiation factor 3 (eIF3)–serves specialized roles to either activate or repress the translation of specific mRNAs. We also found that eIF3 unexpectedly includes its own 5’-m7G cap binding subunit. In this application, we will probe how and when eIF3 carries out its specific regulatory functions. We will use molecular and structural approaches to decipher how eIF3 and trans-acting factors interact with structured RNA elements to regulate the translation of specific mRNAs. We will also examine the role of eIF3 in regulating translation in activated T cells. Finally, we will determine how eIF3 regulation of T cell receptor translation affects T cell development. Answers to these questions will reveal fundamental insights into translational control and will provide a foundation for future engineering of improved cell-based immunotherapies. Protein targets for many human diseases remain “undruggable” due to their underlying biochemical functions and behavior. These limits to small molecule drug discovery hold back the promise of developing affordable therapeutics. We recently showed that small molecules that bind the ribosome can selectively stall the translation of human proteins, revealing an entirely new mechanism of action that could enable targeting previously “undruggable” proteins. These drug-like compounds directly and selectively modulate the translation of specific nascent polypeptides during translation elongation or termination. We also found these compounds impact ribosome quality control pathways. We will explore whether similar mechanisms are employed by cellular metabolites to regulate the translation of specific mRNAs. We will also map new ribosome quality control pathways that target translation frameshifting, a process sensitive to the mechanisms employed by the drug-like compounds to selectively stall translation. Taken together, these experiments will provide new molecular insights that could aid in the design of new small molecule modulators of human translation.

抽象的 蛋白质合成或翻译将所有生命形式的基因型与表型联系起来。凯特实验室有一个 对蛋白质合成机制的长期兴趣，来自细菌收集的普遍原理 翻译是人类翻译调节的基础。该应用程序解决了基本问题 关于人类的翻译是如何受到监管的。我们建议探索翻译起始的监管 特定的细胞和组织以及影响翻译速度和准确性的翻译延伸机制 核糖体。我们认为这两条广泛的研究路线将带来有关蛋白质合成的许多发现 最终可以用来治疗人类疾病。 真核生物翻译起始的规范机制涉及许多一般翻译起始 因素。我们最近发现其中之一——真核起始因子 3 (eIF3)——具有特殊的作用 激活或抑制特定 mRNA 的翻译。我们还发现 eIF3 意外地包括 它自己的 5’-m7G 帽结合亚基。在此应用中，我们将探讨 eIF3 如何以及何时执行其特定的操作 监管职能。我们将使用分子和结构方法来破译 eIF3 和反式作用如何 因子与结构化 RNA 元件相互作用，调节特定 mRNA 的翻译。我们还将 检查 eIF3 在调节活化 T 细胞翻译中的作用。最后，我们将确定 eIF3 如何 T 细胞受体翻译的调节影响 T 细胞的发育。这些问题的答案将揭晓 对平移控制的基本见解，将为未来改进工程提供基础 基于细胞的免疫疗法。 由于其潜在的生化功能，许多人类疾病的蛋白质靶点仍然“无法成药” 和行为。小分子药物发现的这些限制阻碍了开发负担得起的药物的承诺 疗法。我们最近表明，结合核糖体的小分子可以选择性地阻止 人类蛋白质的翻译，揭示了一种全新的作用机制，可以实现靶向 以前“不可成药”的蛋白质。这些药物样化合物直接并选择性地调节翻译 翻译延伸或终止期间特定新生多肽的变化。我们还发现了这些化合物 影响核糖体质量控制途径。我们将探讨是否采用类似的机制 细胞代谢调节特定 mRNA 的翻译。我们还将绘制新核糖体质量图 以翻译移码为目标的控制途径，这是一个对翻译框架所采用的机制敏感的过程 类药物化合物选择性地阻止翻译。总而言之，这些实验将提供新的 分子见解可以帮助设计新的人类翻译小分子调节剂。