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(申请人提供)：帕金森氏病(PD)是美国第二常见的神经退行性疾病。最近，几个常染色体隐性遗传性青少年帕金森综合征(AR-JP)家族被证明携带一种名为Parkin的基因突变，这是一种泛素E3连接酶。NINDS最近确定Parkin和相关蛋白质为深入研究的目标(RFA NS-01-005)。在这项提议中，我们将重点放在Ariadne上，它是已知的与Parkin最接近的结构同源。Ariadne是一种特性不佳的蛋白质，其整体结构组织与Parkin相似。Ariadne和Parkin都与相同的泛素E2酶结合，而在果蝇中，Ariadne的丧失会导致生存能力下降和静止性震颤。在初步的研究中，我们已经证明了Ariadne和Parkin具有重要的结构和功能特性。在这个建议中，我们将使用生化、分子、细胞和遗传学方法来检验Ariadne和Parkin是：1)结构和功能同系物；2)参与路易体的形成；以及3)保护体外和体内的细胞免受伤害性刺激。这项工作可能使我们：1)解释为什么Parkin的全球功能丧失突变的表型效应主要限于多巴胺能神经元；2)验证路易体样胞浆包涵体反映了细胞的适应性反应；和3提供了对这类蛋白质的功能的洞察，这可以增强我们对细胞中Parkin功能的理解。