Defining the role for Gle1 in the fetal lethal motor neuron disease LCCS1

定义 Gle1 在胎儿致死性运动神经元疾病 LCCS1 中的作用

• 批准号: 8061325
• 负责人: Andrew William Folkmann
• 金额: $ 2.64万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8061325
• 关键词: 3' Splice Site; ATP phosphohydrolase; Alleles; Amino Acids; Anterior; Back; Binding; Biochemical; Biological Assay; Biological Models; Birth; Boxing; Cell Line; Cell Nucleus; Cell physiology; Cellular biology; Contracture; Coupled; Coupling; Cytoplasm; DNA; Defect; Development; Disease; Dynein ATPase; Experimental Designs; Experimental Models; Fellowship; Finland; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genetic Translation; Genomics; Goals; Horns; Human; Incidence; Introns; Kinesin; Link; Macromolecular Complexes; Mammalian Cell; Messenger RNA; Metabolism; Modeling; Molecular; Molecular Motors; Motor Activity; Motor Neuron Disease; Motor Neurons; Mutation; Neurons; Nuclear; Nuclear Export; Nuclear Pore Complex; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Phytic Acid; Play; Plus End of the Microtubule; Positioning Attribute; Process; Proteins; RNA; RNA Processing; RNA Transport; Role; Sequence Analysis; Spinal Cord; Syndrome; Testing; Time; Tissues; Translation Initiation; Translation Process; Translations; Work; Yeasts; cell growth regulation; dynactin; fetal; genetic linkage; mutant; novel; research study; small molecule; trafficking

DESCRIPTION (provided by applicant): Nuclear export of messenger RNAs (mRNAs) and their subsequent translation are essential steps in the gene expression pathway and impact all aspects of cell physiology. There is a growing body of evidence suggesting that dysfunctional mRNA trafficking contributes directly to motor neuron diseases (MND). This is further supported by the recent genetic linkage of mutant human GLE1 alleles to the fetal motor neuron disease: Lethal Congenital Contracture Syndrome (LCCS-1). Gle1 is an essential mRNA export factor, conserved from yeast to humans. Yeast Gle1 plays an important role in both translation initiation and termination. Sequence analysis of genomic DNA from LCCS-1 cases identified a mutation that generates an illegitimate splice acceptor site within the third intron of GLE1. This specific mutation (FINMajor) results in an insertion of three amino acid residues (PFQ) in the amino-terminal domain of hGle1. In preliminary studies a novel function for hGle1 in cytoplasmic mRNA trafficking has been found. Strikingly, the FINMajor mutation may disrupt hGle1's function in trafficking. The goal of this proposal is to elucidate the functional consequence of the FINMajor mutant protein in mammalian cells. The two aims proposed will define the molecular consequence of the FINMajor mutation. In Aim I, studies will be done to define hGle1 as a component of the cytoplasmic mRNA trafficking machinery. In Aim II, a novel hGle1 knockdown, and add-back model system will be developed. With this experimental model, cell biology and biochemical assays will be used to investigate if the FINMajor mutant protein disrupts mRNA trafficking. The long-term goal is to molecularly define the neuronal role for hGle1 in the pathogenesis of the deleterious disease LCCS-1 PUBLIC HEALTH RELEVANCE: Gle1 is positioned to regulate multiple steps during mRNA metabolism and gene expression. By defining the molecular functional consequence of the FINMajor mutation, we will elucidate the importance of RNA metabolism in motor neuron disease.

描述（由申请人提供）：信使rna （mrna）的核输出及其随后的翻译是基因表达途径的重要步骤，影响细胞生理的各个方面。越来越多的证据表明，功能失调的mRNA运输直接导致运动神经元疾病（MND）。最近突变的人类GLE1等位基因与胎儿运动神经元疾病：致死性先天性挛缩综合征（LCCS-1）的遗传联系进一步支持了这一点。Gle1是一个重要的mRNA输出因子，从酵母到人类都是保守的。酵母Gle1在翻译起始和终止过程中都起着重要的作用。对LCCS-1病例的基因组DNA序列分析发现，GLE1的第三内含子中存在一个突变，该突变产生了一个非法的剪接受体位点。这种特异性突变（FINMajor）导致hGle1的氨基末端区域插入三个氨基酸残基（PFQ）。在初步研究中发现了hGle1在细胞质mRNA转运中的新功能。引人注目的是，FINMajor突变可能会破坏hGle1在运输中的功能。本研究的目的是阐明FINMajor突变蛋白在哺乳动物细胞中的功能后果。提出的两个目标将定义FINMajor突变的分子后果。在Aim I中，研究将确定hGle1作为细胞质mRNA运输机制的一个组成部分。在Aim II中，将开发一种新的hGle1击倒和添加回模型系统。有了这个实验模型，细胞生物学和生化分析将用于研究FINMajor突变蛋白是否会破坏mRNA的运输。长期目标是从分子上确定hGle1在有害疾病LCCS-1发病机制中的神经元作用