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

基本信息

批准号：
8411599
负责人：
Andrew William Folkmann
金额：
$ 2.69万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
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/8411599
关键词：
3&apos 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 public health relevance research study small molecule trafficking

项目摘要

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

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中的重要步骤，并影响细胞生理学的各个方面。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：致命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 是一种重要的 mRNA 输出因子，从酵母到人类都是保守的。酵母 Gle1 在翻译起始和终止中发挥着重要作用。对 LCCS-1 病例的基因组 DNA 进行序列分析，发现了一种突变，该突变在 GLE1 的第三个内含子内产生非法剪接受体位点。这种特定突变 (FINMajor) 导致在 hGle1 的氨基末端结构域中插入三个氨基酸残基 (PFQ)。初步研究发现 hGle1 在细胞质 mRNA 运输中的新功能。引人注目的是，FINMajor 突变可能会破坏 hGle1 的运输功能。该提案的目的是阐明 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。在目标 I 中，将进行研究以将 hGle1 定义为细胞质 mRNA 运输机制的一个组成部分。在 Aim II 中，将开发一种新颖的 hGle1 击倒和回加模型系统。通过该实验模型，将使用细胞生物学和生化检测来研究 FINMajor 突变蛋白是否会破坏 mRNA 运输。长期目标是从分子水平上确定 hGle1 在有害疾病 LCCS-1 发病机制中的神经元作用