Regulation of the SCA3 Disease Gene and Its Protein

SCA3疾病基因及其蛋白的调控

• 批准号: 6622450
• 负责人: SARAH J BERKE
• 金额: $ 2.44万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6622450
• 关键词: cerebellar ataxia /dyskinesia; cysteine endopeptidases; enzyme activity; gene induction /repression; genetic transcription; glutamine; guanine nucleotide binding protein; homopeptide; nerve /myelin protein; neurobiology; neurogenetics; proteasome; proteolysis; tissue /cell culture

DESCRIPTION (provided by applicant): The aim of this proposal is to investigate the transcriptional and posttranslational regulation of the disease protein causing the polyglutamine (polygln) disease spinocerebellar ataxia type-3 (SCA3), a protein known as ataxin-3. Polygln diseases are dominantly inherited, ultimately fatal neurodegenerative diseases, for which there is no effective therapy. A unifying pathological feature of polygln diseases is intracellular aggregation of the disease protein, arguing that precise control of protein homeostasis is vital to neuronal survival. Therefore, therapeutic strategies should include efforts to: 1) enhance degradation of the disease protein; 2) boost the protein surveillance machinery of the neuron; and 3) modulate disease gene expression. Toward this goal, the specific aims are to determine the cellular pathways controlling ataxin-3 degradation, and to define the transcriptional regulation of the SCA3 disease gene, MJD1. Specifically, studies in aim 1 hope to demonstrate that ataxin-3 is a normal target for proteasome and caspase proteolysis, and determine whether the proteasomal activity is inhibited by accumulated, mutant polygln protein. Studies in aim 2 will test the hypothesis that the Ras/MEK pathway regulates MJD1 gene transcription. A variety of cellular and biochemical methods will be employed to answer these questions including metabolic pulse-chase assays, coimmunoprecipitation experiments, immunoblots, site-directed mutagenesis, cell synchronization, and promoter analysis. The studies proposed here will further our understanding of ataxin-3 production and degradation, perhaps leading to the development of therapies for these currently untreatable diseases.

描述(由申请人提供)： 这项建议的目的是调查转录和 引起多聚谷氨酰胺的疾病蛋白的翻译后调节 (Polygln)病脊髓小脑性共济失调3型(SCA3)，一种被称为 紫杉素-3。多发性疾病主要是遗传的，最终是致命的 神经退行性疾病，没有有效的治疗方法。一个统一的 多发性神经病的病理特征是细胞内聚集 疾病蛋白质，认为精确控制蛋白质动态平衡至关重要 为了神经元的生存。因此，治疗策略应该包括努力 为了：1)促进疾病蛋白质的降解；2)促进蛋白质 神经元的监测机制；以及3)调节疾病基因的表达。 为了达到这个目标，具体的目标是确定细胞的路径 控制ataxin-3的降解，并确定转录调控 SCA3病基因MJD1.具体地说，Aim 1的研究希望 证明ataxin-3是蛋白酶体和caspase的正常靶点 蛋白分解，并确定蛋白酶体活性是否被抑制 积累的、突变的Polygln蛋白。《目标2》中的研究将检验这一假设 Ras/MEK途径调控MJD1基因转录。各种各样的 我们将使用细胞和生化方法来回答这些问题。 包括代谢脉冲追逐试验，免疫共沉淀实验， 免疫印迹、定点突变、细胞同步化和启动子 分析。这里提出的研究将进一步加深我们对ataxin-3的理解 产生和降解，可能导致治疗方法的发展 这些目前无法治愈的疾病。