Translational Control by the Fragile X Mental Retardation Protein

脆性 X 智力迟钝蛋白的翻译控制

基本信息

批准号：
9199419
负责人：
SIMPSON JOSEPH
金额：
$ 35.48万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2019-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9199419
关键词：
American Amino Acids Animal Model Behavioral Binding Binding Proteins Biochemical Biological Assay Brain Cessation of life Childhood Code Complex Cryoelectron Microscopy Data Development Disease Drosophila FMR1 protein Drosophila genus Drosophila inturned protein Drug Targeting Exhibits FMR1 FMRP FXR1 gene FXR2 gene Facioscapulohumeral Muscular Dystrophy Fluorescence Fragile X Syndrome G-Quartets Genes Genetic Genetic Transcription Goals Human Human Cell Line In Vitro Inherited Intellectual functioning disability Laboratories Mammals Mental Retardation Messenger RNA Molecular Monitor Mus Muscle Development Muscular Dystrophies Myocardium Patients Phenotype Prevalence Protein Biosynthesis Proteins RNA RNA-Binding Proteins Regulation Reporter Resolution Ribosomes Role Seizures Skeletal Muscle Specificity Structure System Techniques Transgenic Organisms Translations autistic behaviour base biophysical techniques experimental study fetal fly in vivo insight interest mRNA Expression male myogenesis nervous system disorder novel therapeutics paralogous gene protein complex protein function public health relevance tool translation factor

项目摘要

DESCRIPTION (provided by applicant) Fragile X syndrome is a disease that afflicts about 100,000 Americans and about 3 million people worldwide, resulting in intellectual disability, childhood seizures, and autistic behavior i the patients. The disease is caused by the transcriptional silencing of the fragile X mental retardation 1 gene (FMR1). FMR1 gene codes for a RNA-binding protein, the fragile X mental retardation protein (FMRP), which is highly expressed in the brain and is essential for the normal development of the brain. Mammals have two autosomal paralogs of FMRP designated as fragile X related 1 and 2 (FXR1 and FXR2) proteins. FXR1 is essential for myogenesis and the altered expression of FXR1 causes facioscapulohumeral muscular dystrophy, the most prevalent form of muscular dystrophy. Inactivation of FXR2 does not cause a disease in humans; however, loss of both FMRP and FXR2 results in a more severe form of FXS in mice and Drosophila. Thus, FMRP and FXR2 appear to have overlapping functions in the brain, whereas FXR1 is more critical for muscle development. FMRP, FXR1 and FXR2 have been implicated in regulating the translation of several mRNAs. However, the precise mechanism by which these proteins regulate the expression of these mRNAs is unknown. The goal of the proposed study is to understand the molecular mechanism underlying the regulation of protein synthesis by FMRP, FXR1 and FXR2. We have strong initial results showing that FMRP can bind directly to the 80S ribosome to regulate protein synthesis. In Specific Aim 1, we will dissect the mechanism of translational control by Drosophila FMRP. In Specific Aim 2, we will focus on the mechanism of translational control by human FMRP, FXR1 and FXR2. For both specific aims, we will use a robust in vitro translation system and quantitative biophysical methods that have been developed in our laboratory, and in vivo studies, using human cell lines and transgenic Drosophila. These functional analyses, in conjunction with the proposed high-resolution cryo-electron microscopy, will significantly advance our understanding of the molecular mechanism used by FMRP, FXR1 and FXR2 to regulate protein synthesis. Results of these studies will provide useful insights in identifying potential drug targets to treat fragile X syndrome and facioscapulohumeral muscular dystrophy.

描述（由申请人提供）脆弱的X综合征是一种疾病，在全球范围内遭受约100,000名美国人和约300万人的疾病，导致智力残疾，儿童癫痫发作和患者加速行为。该疾病是由脆弱的X智力低下1基因（FMR1）的转录沉默引起的。 FMR1基因编码RNA结合蛋白，即脆弱的X智力低下蛋白（FMRP），该蛋白在大脑中高度表达，对于大脑的正常发育至关重要。哺乳动物具有指定为脆弱X相关1和2（FXR1和FXR2）蛋白的FMRP的两个常染色体旁系同源物。 FXR1对于肌发生至关重要，FXR1的表达改变会导致肌肉旋转肌肉营养不良，这是最普遍的肌肉营养不良形式。 FXR2的失活不会引起人类疾病；然而，FMRP和FXR2的丧失导致小鼠和果蝇中更严重的FXS形式。这是FMRP和FXR2在大脑中似乎具有重叠的功能，而FXR1对于肌肉发育更为重要。 FMRP，FXR1和FXR2已在调节几个mRNA中实施。但是，这些蛋白质调节这些mRNA表达的精确机制尚不清楚。拟议的研究的目的是了解FMRP，FXR1和FXR2调节蛋白质合成的基础机制。我们的初始结果表明，FMRP可以直接与80S核糖体结合以调节蛋白质合成。在特定目标1中，我们将剖析果蝇FMRP翻译控制的机制。在特定的目标2中，我们将重点介绍人FMRP，FXR1和FXR2的翻译控制机制。对于这两个特定目标，我们将使用在我们的实验室中开发的强大体外翻译系统和定量生物物理方法，并使用人类细胞系和转基因果蝇进行体内研究。这些功能分析与提议的结合。高分辨率的冷冻电子显微镜将显着提高我们对FMRP，FXR1和FXR2用于调节蛋白质合成的分子机制的理解。这些研究的结果将提供有用的见解，以识别潜在的药物靶标以治疗脆弱的X 综合征和Facioscapulohumeral肌肉营养不良。