Do Ariadne and Parkin Share Redundant Function

Ariadne 和 Parkin 共享冗余功能吗

• 批准号: 6572482
• 负责人: LAWRENCE M SCHWARTZ
• 金额: $ 33.92万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-15 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6572482
• 关键词: Drosophilidae; Lewy body; Parkinson's disease; SDS polyacrylamide gel electrophoresis; cell differentiation; cell line; cytoprotection; dopamine; flow cytometry; genetically modified animals; myogenesis; neural degeneration; neurogenetics; parkin gene /protein; protein structure; protein structure function; ubiquitin; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is the second most common neurodegenerative disorder in the United States. Recently, several families with Autosomal Recessive Juvenile Parkinsonism (AR-JP) have been demonstrated to carry mutations in a gene termed Parkin, a ubiquitin E3 ligase. NINDS recently identified Parkin and related proteins as targets for intensive investigation (RFA NS-01-005). In this proposal, we focus on Ariadne, the closest known structural homolog of Parkin. Ariadne is a poorly characterized protein with a similar overall structural organization to Parkin. Both Ariadne and Parkin bind to the same ubiquitin E2 enzymes and in Drosophila, loss of Ariadne results in reduced viability and resting tremors. In preliminary studies, we have demonstrated that Ariadne and Parkin share significant structural and functional properties.In this proposal, we will use biochemical, molecular, cellular and genetic methods to test thehypotheses that Ariadne and Parkin are: 1) structural and functional homologs; 2) participate in Lewy Body formation; and 3) protect cells both in vitro and in vivo from noxious stimuli.This work may allow us to: 1) explain why the phenotypic effects of global loss-of-function mutations in Parkin are largely restricted to dopaminergic neurons; 2) verify that Lewy Body-like cytoplasmic inclusions reflect an adaptive response by cells; and 3 provide insight into the functioning of this class of proteins which can enhance our understanding of Parkin function in cells.

描述（由申请人提供）：帕金森病（PD）是美国第二常见的神经退行性疾病。最近，几个患有常染色体隐性青少年帕金森症 (AR-JP) 的家族已被证明携带称为 Parkin（一种泛素 E3 连接酶）的基因突变。 NINDS 最近将 Parkin 和相关蛋白确定为深入研究的目标 (RFA NS-01-005)。在本提案中，我们重点关注 Ariadne，它是已知与 Parkin 最接近的结构同系物。 Ariadne 是一种特征很少的蛋白质，其总体结构组织与 Parkin 相似。 Ariadne 和 Parkin 都与相同的泛素 E2 酶结合，在果蝇中，Ariadne 的缺失会导致活力降低和静息性震颤。在初步研究中，我们已经证明Ariadne和Parkin具有重要的结构和功能特性。在本提案中，我们将利用生化、分子、细胞和遗传学方法来检验Ariadne和Parkin的假设：1）结构和功能同源物； 2）参与路易体形成； 3）保护体外和体内细胞免受有害刺激。这项工作可能使我们能够：1）解释为什么Parkin中整体功能丧失突变的表型效应主要限于多巴胺能神经元； 2）验证类路易体细胞质内含物反映细胞的适应性反应； 3 提供了对此类蛋白质功能的深入了解，这可以增强我们对细胞中 Parkin 功能的理解。