Functional analysis of Attractin-Mahogunin signaling

Attractin-Mahogunin 信号传导的功能分析

基本信息

批准号：
7249350
负责人：
Teresa M Gunn
金额：
$ 35.85万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-06-01 至 2009-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7249350
关键词：
Adult Aging Alleles Alternative Splicing Alzheimer&apos s Disease Animal Model Antibodies Biochemical Biological Assay Brain CNS degeneration Cells Cessation of life Color Complex Defect Disease Evaluation Fingers Gene Expression Genes Goals Homologous Gene Immunohistochemistry In Vitro Laboratories Lead Mammalian Cell Mediating Methods Modeling Mus Mutant Strains Mice Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Parkinson Disease Pathway interactions Pattern Phenotype Physiological Pigmentation physiologic function Pigments Protein Isoforms Proteins Research Role Signal Pathway Signal Transduction Testing Tissues Transgenic Mice Two-Hybrid System Techniques Ubiquitin Ubiquitination Upper arm Yeasts attractin protein base gene function in vivo mahogunin protein mutant neuron loss parkin gene/protein protein aggregate ubiquitin-protein ligase yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Neurodegenerative disorders such as Alzheimer's and Parkinson's disease are common and devastating diseases, but the mechanism leading to neuronal death is not well understood. Mice with mutations in the Attractin (Atrn) and Mahogunin (Mgn) genes develop progressive spongy degeneration of the brain and thus represent newly recognized models of neurodegeneration. These mutants also have a coat color phenotype due to a defect in a pigment-cell specific pathway. The similarity between Atrn and Mgn mutants for two unrelated phenotypes suggests a common function for these genes. While the role of Atrn in neurons is unclear, Mgn encodes a RING-finger containing protein that acts as a ubiquitin ligase (E3) in vitro. Accumulation of ubiquitinated protein aggregates is a hallmark of many neurodegenerative disorders and mutations in another E3, Parkin, cause a familial form of Parkinson's disease. Thus, we hypothesize that defects in ubiquitin-mediated degradation of specific target proteins lead to neuronal death in Atrn and Mgn mutants. The long-term goal of research in the Gunn laboratory is to determine how. The short term goals are to test the following hypotheses: 1) that Mgn functions as an E3 in vivo, by testing whether the RING domain of Mgn (required for E3 activity in vitro) is essential for normal Mgn function in vivo, and by identifying Mgn-interacting proteins using a yeast two hybrid assay and using biochemical assays to test whether these proteins are targeted by Mgn for ubiquitin-mediated decay and accumulate in the brains of Mgn mutant mice. 2) that Mgn and Atrn act in the same pathway, using marker gene expression, western analysis and/or immunohistochemistry to examine Mgn levels and localization in Atrn mutants and determine whether proteins identified as Mgn targets for ubiquitination accumulate in the brains of Atrn mutant mice. 3) that a newly discovered Atrn homolog, Lurin, signals through the Mgn pathway, by generating mice lacking Lurin and examining their phenotype (including accumulation of Mgn targets) on normal and Atrn null backgrounds. As Mgn mutants display some phenotypes not observed in Atrn mutants, Lrn may compensate for loss of Atrn in some tissues and loss of both Lrn and Atrn may recapitulate the full Mgn phenotype.

描述（由申请人提供）：阿尔茨海默病和帕金森病等神经退行性疾病是常见且具有破坏性的疾病，但导致神经元死亡的机制尚不清楚。 Attractin (Atrn) 和 Mahogunin (Mgn) 基因突变的小鼠会出现大脑进行性海绵状变性，因此代表了新近公认的神经变性模型。由于色素细胞特异性途径的缺陷，这些突变体还具有毛色表型。两个不相关表型的 Atrn 和 Mgn 突变体之间的相似性表明这些基因具有共同的功能。虽然 Atrn 在神经元中的作用尚不清楚，但 Mgn 编码一种含有环指的蛋白质，该蛋白质在体外充当泛素连接酶 (E3)。泛素化蛋白聚集体的积累是许多神经退行性疾病的标志，另一种 E3 蛋白 Parkin 的突变会导致家族性帕金森病。因此，我们假设泛素介导的特定靶蛋白降解的缺陷导致 Atrn 和 Mgn 突变体中的神经元死亡。冈恩实验室研究的长期目标是确定如何实现。短期目标是检验以下假设： 1) Mgn 在体内起到 E3 的作用，通过测试 Mgn 的 RING 结构域（体外 E3 活性所需）对于体内正常 Mgn 功能是否至关重要，并使用酵母两种杂交测定法鉴定 Mgn 相互作用蛋白，使用生化检测来测试这些蛋白质是否是 Mgn 的靶标，进行泛素介导的衰变并在 Mgn 突变小鼠的大脑中积累。 2) Mgn和Atrn作用于相同的途径，使用标记基因表达、蛋白质印迹分析和/或免疫组织化学来检查Atrn突变体中的Mgn水平和定位，并确定被鉴定为泛素化Mgn靶标的蛋白质是否在Atrn突变小鼠的大脑中积累。 3) 新发现的 Atrn 同源物 Lurin 通过 Mgn 途径产生小鼠来发出信号缺乏 Lurin 并在正常和 Atrn 无效背景下检查其表型（包括 Mgn 靶标的积累）。由于 Mgn 突变体表现出一些在 Atrn 突变体中未观察到的表型，Lrn 可能会补偿某些组织中 Atrn 的损失，并且 Lrn 和 Atrn 的损失可能会重现完整的 Mgn 表型。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genetic analysis of attractin homologs.

吸引素同系物的遗传分析。

DOI：
发表时间：
2007-12
期刊：
影响因子：
0
作者：
Walker, Will P;Aradhya, Swaroop;Hu, Che;Shen, Shiliang;Zhang, Wei;Azarani, Arezou;Lu, XinYun;Barsh, Gregory S;Gunn, Teresa M
通讯作者：
Gunn, Teresa M

Transgenic analysis of the physiological functions of Mahogunin Ring Finger-1 isoforms.

Mahogunin Ring Finger-1 亚型生理功能的转基因分析。