FUNCTION OF THE DRPLA GENE PRODUCT ATROPHIN

这个项目的目标是阐明阿托啡的功能。 蛋白质，其突变是神经退行性疾病的基础 齿状回-苍白球萎缩(DRPLA)。这些突变是 多聚谷氨酰胺重复序列(由CAG三核苷酸编码)的扩增 在蛋白质中重复)，使DRPLA成为几个 众所周知的“谷氨酰胺重复紊乱症”。在这组神经退行性疾病中 疾病，多聚谷氨酰胺重复序列在许多疾病中的扩张 不同的蛋白质赋予不同的疾病表型。但是，在 尽管差异细胞死亡的模式相对有限 在DRLA和其他疾病中，基因产物广泛存在 分布于中枢神经系统和外周组织， 它们的正常功能和致病机制在很大程度上 未知。DRPLA基因产物阿托芬具有以下几个优点 用于研究的模型蛋白质；它是亲水性的，大小适中， 包含许多有趣的多肽主题，这些主题可能很重要 在它的功能上。这项研究将考察阿托啡的功能 通过四个具体目标。首先，会产生抗体来对抗 纯化的萎缩蛋白、细菌融合蛋白和小分子合成的 多肽。这些试剂将被用来探测区域、细胞、 以及脑内萎缩蛋白的亚细胞定位。这次会议的重点是 然而，这个项目将是识别相互作用的蛋白质 用阿托芬、酵母菌双杂交进行筛选和亲和 提纯技术。聚谷氨酰胺的膨化效果 然后将对任何已确定的交互进行重复评估。一个焦点 关于蛋白质-蛋白质相互作用的研究在评估 阿托芬的功能，如这种相互作用中由于 多聚谷氨酰胺重复序列扩增可能参与了糖尿病的发病机制 DR解放军。最后，差异选择性剪接的效果 我们将研究萎缩蛋白基因对萎缩蛋白-蛋白质相互作用的影响。 遗传性疾病，如DRPLA和其他谷氨酰胺重复 神经学家对疾病特别感兴趣，因为 这些遗传性疾病中神经退行性变的机制 可能揭示了在更常见的获得性疾病中类似的机制 神经退行性疾病，如阿尔茨海默病、肌营养不良 侧索硬化症和帕金森氏症。 在本研究过程中，由Morgan Sheng MBBS，PhD， 应聘者希望接受尖端技术方面的培训 分子遗传学和蛋白质生物化学将使他能够 在分子神经学方面追求富有成效的职业生涯。

The goal of this project is to elucidate the function of the atrophin protein, mutations of which underlie the neurodegenerative disease dentatorubral-pallidoluysian atrophy (DRPLA). These mutations are expansions of a polyglutamine repeat motif (coded by CAG trinucleotide repeats in the mRNA) within the protein, making DRPLA one of several known "glutamine repeat disorders." In this group of neurodegenerative diseases, an expansion of a polyglutamine repeat in a number of different proteins confers the various disease phenotypes. However, in spite of the relatively restricted patterns of differential cell death in DRPLA and the other disorders, the gene products are widely distributed in both the central nervous system and peripheral tissues, and both their normal functions and mechanisms of disease are largely unknown. The DRPLA gene product atrophin has a number of advantages as a model protein for study; it is hydrophilic, of moderate size, and contains a number of intriguing peptide motifs which may be important in its function. This study will examine the function of atrophin through four specific aims. First, antibodies will be generated against purified atrophin, bacterial fusion proteins, and small synthetic peptides. These reagents will be used to probe the regional, cellular, and subcellular localizations of atrophin in brain. The focus of the project, however, will be the identification of proteins that interact with atrophin, using yeast two hybrid screening and affinity purification techniques. The effect of expansions of the polyglutamine repeat on any identified interactions will then be assessed. A focus on protein-protein interactions is critical while evaluating the function of atrophin, as alterations in such interactions due to the polyglutamine repeat expansion may be involved in the pathogenesis of DRPLA. Finally, the effects of differential alternative splicing of the atrophin gene on atrophin protein-protein interactions will be examined. Hereditary disorders such as DRPLA and the other glutamine repeat diseases are of particular interest to neurologists, as identification of the mechanism of neurodegeneration in these hereditary disorders will likely shed light on similar mechanisms in the more common acquired neurodegenerative diseases such as Alzheimer's disease, amyotrophic lateral sclerosis, and Parkinson's disease. During the course of this research, sponsored by Morgan Sheng MBBS, PhD, the candidate expects to be trained in the cutting edge techniques of molecular genetics and protein biochemistry that will empower him to pursue a productive career in molecular neurology.