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Regulation of synaptic proteasome activity by the dynactin complex

dynactin 复合物对突触蛋白酶体活性的调节

基本信息

批准号：
8420454
负责人：
Benjamin Arthur Eaton
金额：
$ 30.72万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8420454
关键词：
Affect Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Axonal Transport Binding Binding Proteins Biochemical Biological Assay Cell membrane Cellular Assay Cellular biology Complex Data Deposition Development Disease Drosophila genus Dynein ATPase Enhancers Event Family Fluorescence Frontotemporal Dementia Functional disorder Genes Genetic Genetic Screening Genetic Techniques Glues Goals Golgi Apparatus Growth Guanosine Triphosphate Phosphohydrolases Health Homologous Gene Human Human Pathology Image Impairment In Vitro Investigation Link Lipid Binding Maintenance Mediating Mediation Membrane Modeling Molecular Molecular Genetics Monomeric GTP-Binding Proteins Mus Mutation Nerve Nerve Degeneration Nervous System Physiology Nervous system structure Neurodegenerative Disorders Neurons Neurophysiology - biologic function Onset of illness Parkinson Disease Pathogenesis Pathway interactions Phenocopy Process Proteasome Inhibition Proteins Reagent Regulation Reporter Resolution Role Sequence Analysis Signal Transduction Structure Synapses System Tertiary Protein Structure Vesicle base design dynactin fly human disease in vivo interest multicatalytic endopeptidase complex mutant nervous system disorder neurodegenerative phenotype neurological pathology neurotransmission novel novel strategies overexpression presynaptic progressive neurodegeneration protein aggregate protein complex protein degradation response rho synaptic function tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Pinpointing the underlying cause of many neurodegenerative diseases has been difficult due to the complexity of neuronal cellular biology and the lack of basic information about impaired mechanisms that contribute to disease. Mutations in the human DCTN1 gene, which encodes a component of the dynactin complex, have been strongly linked to both familial and sporadic cases of amyotrophic lateral sclerosis (ALS). Many of the neurodegenerative phenotypes associated with human disease have been recapitulated in flies and mice harboring dynactin complex mutations demonstrating a conserved pathogenesis in dynactin complex mutants. To clarify the mechanisms that contribute to the pathogenesis of neurodegeneration observed in dynactin complex mutants, especially the degeneration of synaptic contacts, we will use a combination of forward genetic screens and quantitative cellular assays to identify and characterize important modifiers of dynactin complex function at the synapse. We believe that this approach has the potential to identify and describe new genes and pathways required within the nervous system for normal synaptic growth, synapse stabilization, and function. A genetic screen designed to specifically identify genetic modifiers of the Drosophila DCTN1 homolog, glued, within the nervous system has identified the Drosophila homologue of the Arfaptin2 gene, Darfaptin2 (Darf2). We find that Darf2 is expressed in motorneurons and required for normal synaptic growth. The goal of this proposal is to investigate the hypothesis that Darfaptin2 (Darf2) represents a novel component of the dynactin complex required for normal synaptic growth, stabilization, and neurotransmission. Models of Arfaptin2 function include the mediation of cross-talk between Rho-like GTPases and Arf family GTPases during vesicle formation, and the regulation of proteasome activity within neurons. Using standard genetic techniques and synaptic analyses, we will first define the role of Darf2 in the regulation of synaptic growth and synapse stabilization. This will include the biochemical analysis of its association with the dynactin complex in the nervous system (Aim1). We will further investigate the role of Darf2 in the nervous system using a structure-function approach to define protein domains required for normal Darf2 activity. This will include determining the signaling context for Darf2 during synapse growth and stabilization (Aim2). Finally, we are developing reagents and assays to directly investigate the regulation of proteasome function in the nerve terminal by the dynactin complex, Darf2, and during synapse retraction (Aim3). It is expected that these studies will reveal a novel regulatory mechanism during synaptic growth, stabilization, and neurotransmission that will have important implications for the pathogenesis of late onset neurological diseases.

描述（由申请人提供）：由于神经细胞生物学的复杂性和缺乏导致疾病的受损机制的基本信息，确定许多神经退行性疾病的潜在原因一直很困难。人类DCTN1基因的突变与家族性和散发性肌萎缩性侧索硬化症（ALS）密切相关，DCTN1基因编码dynactin复合物的一个组成部分。许多与人类疾病相关的神经退行性表型已在携带动蛋白复合物突变的果蝇和小鼠中重现，表明动蛋白复合物突变具有保守的发病机制。为了阐明在dynactin复合物突变体中观察到的神经退行性变的发病机制，特别是突触接触的退行性变，我们将使用正向遗传筛选和定量细胞分析相结合的方法来识别和表征突触中dynactin复合物功能的重要修饰因子。我们相信这种方法有潜力识别和描述神经系统中正常突触生长、突触稳定和功能所需的新基因和途径。一种专门用于鉴定果蝇DCTN1同源基因修饰因子的基因筛选，粘在神经系统内，已经鉴定出果蝇的Arfaptin2同源基因Darfaptin2 （Darf2）。我们发现Darf2在运动神经元中表达，是正常突触生长所必需的。本研究的目的是研究Darfaptin2 （Darf2）是否代表了正常突触生长、稳定和神经传递所需的动蛋白复合体的一种新成分。Arfaptin2的功能模型包括在囊泡形成过程中介导rho样gtpase和Arf家族gtpase之间的串扰，以及调节神经元内蛋白酶体的活性。使用标准的遗传技术和突触分析，我们将首先确定Darf2在突触生长和突触稳定调控中的作用。这将包括其与神经系统动蛋白复合物（Aim1）关联的生化分析。我们将使用结构-功能方法进一步研究Darf2在神经系统中的作用，以确定正常Darf2活性所需的蛋白质结构域。这将包括确定突触生长和稳定期间Darf2的信号环境（Aim2）。最后，我们正在开发试剂和检测方法来直接研究动力蛋白复合物Darf2和突触缩回（Aim3）对神经末梢蛋白酶体功能的调节。这些研究将揭示突触生长、稳定和神经传递过程中的一种新的调控机制，这将对晚发性神经疾病的发病机制产生重要影响。