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

基本信息

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
项目类别：
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&apos 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

项目摘要

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需要细胞命运的决定。在早期脊椎动物发育过程中精确控制特定蛋白质的时间和空间表达。某些成体细胞类型，例如神经系统的细胞，严重依赖于这种受调节的蛋白质表达关于转录后机制，特别是翻译控制。 RNA 结合蛋白 Bicaudal-C (Bicc1) 具有翻译调节功能，对于 Bicc1 是正常脊椎动物发育所需的 RNA 结合蛋白。 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。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。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生物疾病。中心假设是 Bicc1 通过复杂的 RNA 结合结构域选择特定的靶 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该假设基于实验室的广泛研究，重点是定义 Bicc1 如何指导。这项工作揭示了模式生物非洲爪蟾中脊椎动物发育的最早的母体阶段。建立 Bicc1 作为理解 RNA 结合蛋白如何控制 mRNA 翻译的范例直接复杂的细胞命运决定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 结合和翻译调控所需的 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；以及 3. 定义 Bicc1 mRNA 靶点的作用该研究采用了严格的多学科方法来研究脊椎动物发育过程中的细胞命运决定。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。