Regulation of cell fates by the Bicaudal-C translational repressor

Bicaudal-C 翻译阻遏蛋白对细胞命运的调节

• 批准号: 9523681
• 负责人: Michael D Sheets
• 金额: $ 30.76万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-05 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9523681
• 关键词: 3' Untranslated Regions; Address; Adult; Affect; Animal Model; Animals; Area; Binding; Biological; Biological Assay; Cell Fate Control; Cell physiology; Cells; Complex; Coupled; Defect; Development; Developmental Process; Disease; Elements; Embryo; Embryology; Embryonic Development; Eukaryotic Cell; Foundations; Functional disorder; Gene Expression; Genetic Transcription; Genetic Translation; Genome; Goals; Growth and Development function; Health; Human; In Vitro; Individual; Knowledge; Mass Spectrum Analysis; Maternal Messenger RNA; Mediating; Messenger RNA; Modeling; Molecular; Molecular Genetics; Mutation Analysis; Names; Nervous system structure; Nucleotides; Organism; Outcome; Phase; Physiological Processes; Population; Process; Protein Biochemistry; Proteins; Public Health; RNA; RNA Binding; RNA Recognition Motif; RNA Sequences; RNA-Binding Proteins; RNA-Protein Interaction; RNase protection assay; Reporter; Repression; Research; Role; Surface; Testing; Time; Translational Regulation; Translational Repression; Translations; Untranslated Regions; Vertebrates; Work; Xenopus; Xenopus laevis; base; body system; cell fate specification; cell type; citrate carrier; cohort; deep sequencing; flexibility; genetic regulatory protein; in vivo; mRNA Expression; multidisciplinary; mutant; protein crosslink; protein expression; protein function; translation assay

Abstract: Over the course of an animal’s lifetime, cell-fate decisions are continually being made that allow for normal development and growth as well as the health of the adult organism. Cell-fate decisions require precisely controlled temporal and spatial expression of particular proteins. In early vertebrate development and certain adult cell types, such as those of the nervous system, this regulated protein expression relies heavily on post-transcriptional mechanisms, particularly translational control. This proposal focuses on a conserved RNA binding protein named Bicaudal-C (Bicc1) that functions in translational regulation and is essential for normal vertebrate development. While it is established that Bicc1 is an RNA binding protein required for the normal development and health of vertebrates, the cellular and molecular mechanisms by which Bicc1 performs these roles are largely unknown and thus represent a major gap in knowledge. In addition, because several relevant Bicc1 target mRNAs have only recently been identified, their roles in vertebrate development are also unknown, limiting the ability to connect Bicc1’s molecular functions to specific cell-fate decisions. The long-term research goal is to define the molecular mechanisms by which developmentally important RNA binding proteins select their target mRNAs and control mRNA expression to effect specific cell-fate decisions, and to understand how defects in these processes contribute to cell dysfunction and organismal disease. The central hypothesis is that Bicc1 selects particular target mRNAs through a complex RNA binding domain with multiple independent RNA binding surfaces, and regulates translation via additional distinct regions yet to be defined. This hypothesis is based on extensive research from the lab focused on defining how Bicc1 directs the earliest, maternal stages of vertebrate development in the model organism Xenopus laevis. This work has established Bicc1 as a paradigm for understanding how RNA binding proteins control mRNA translation to direct complex cell-fate decisions. Building on extensive conceptual and technical progress over the past decade, the Specific Aims will address the central hypothesis by: 1. Defining elements within Bicc1 target mRNAs required for Bicc1 binding and translational regulation; 2. Determining the regions of Bicc1 that are necessary and sufficient to selectively bind and contact mRNAs and to function in translational regulation. These regions’ roles in embryogenesis will also be examined; and 3. Defining the roles of Bicc1 mRNA targets in cell-fate decisions during vertebrate development. The research employs a rigorous and multidisciplinary strategy incorporating RNA-protein biochemistry, unique translation-reporter assays, genome-enabled approaches, reverse molecular genetics and embryology to define the molecular mechanisms by which the conserved and disease-relevant RNA binding protein Bicc1 directs the earliest cell-fate decisions essential for vertebrate development.

摘要：在动物的一生中，细胞命运的决定不断做出，以允许 正常发育和生长以及成人有机体的健康。细胞命运的决定需要 精确控制特定蛋白质的时间和空间表达。在脊椎动物的早期发育和 某些成体细胞类型，例如神经系统的细胞，这种受调节的蛋白质表达在很大程度上依赖于 转录后机制，特别是翻译控制。该提案重点关注保守的 RNA 结合蛋白 Bicaudal-C (Bicc1) 具有翻译调节功能，对于 脊椎动物的正常发育。虽然已确定 Bicc1 是 RNA 结合蛋白，但 Bicc1 影响脊椎动物正常发育和健康的细胞和分子机制 所扮演的这些角色在很大程度上是未知的，因此代表了知识上的重大差距。另外，因为 几个相关的 Bicc1 靶 mRNA 最近才被鉴定，它们在脊椎动物发育中的作用 也是未知的，限制了将 Bicc1 分子功能与特定细胞命运决定联系起来的能力。这 长期研究目标是确定对发育重要的RNA的分子机制 结合蛋白选择其目标 mRNA 并控制 mRNA 表达以影响特定的细胞命运决定， 并了解这些过程中的缺陷如何导致细胞功能障碍和机体疾病。这 中心假设是 Bicc1 通过复杂的 RNA 结合域选择特定的靶 mRNA 多个独立的RNA结合表面，并通过其他不同的区域调节翻译 定义的。该假设基于实验室的广泛研究，重点是定义 Bicc1 如何指导 模式生物非洲爪蟾中脊椎动物发育的最早的母体阶段。这部作品有 建立 Bicc1 作为理解 RNA 结合蛋白如何控制 mRNA 翻译的范例 直接决定复杂的细胞命运。建立在过去广泛的概念和技术进步的基础上 十年内，具体目标将通过以下方式解决中心假设： 1. 定义 Bicc1 目标中的要素 Bicc1 结合和翻译调节所需的 mRNA； 2. 确定 Bicc1 的区域 选择性结合和接触 mRNA 并在翻译调节中发挥作用是必要和充分的。 还将检查这些区域在胚胎发生中的作用； 3. 定义 Bicc1 mRNA 靶标的作用 脊椎动物发育过程中细胞命运的决定。该研究采用了严格的、多学科的 结合RNA-蛋白质生物化学、独特的翻译报告基因检测、基因组支持的策略 方法，逆向分子遗传学和胚胎学来定义分子机制 保守且与疾病相关的RNA结合蛋白Bicc1指导最早的细胞命运决定，这对细胞命运至关重要 脊椎动物的发育。