The PPD (Parkin-PINK1-DJ-1) E3 Ligase Complex and Neurodegeneration

PPD (Parkin-PINK1-DJ-1) E3 连接酶复合物与神经退行性变

基本信息

批准号：
7883262
负责人：
ZHUOHUA ZHANG
金额：
$ 41.36万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7883262
关键词：
Antioxidants Cells Complex Development Disease Disease Progression Drosophila genus Genes Human Knowledge LRRK2 gene Levodopa Lewy Bodies Link Mediating Modeling Molecular Movement Movement Disorders Mus Muscle Rigidity Mutation Nerve Degeneration Neurons Oxidative Stress PINK1 gene PPD E3 Ligase Complex Parkinson Disease Parkinsonian Disorders Pathway interactions Patients Phosphorylation Phosphotransferases Play Predisposition Protein-Serine-Threonine Kinases Proteins RNA Interference Regulation Research Research Personnel Role Sequence Analysis Small Interfering RNA Substantia nigra structure Technology Testing Tremor Ubiquitination Vitamin E age related alpha synuclein base dopaminergic neuron early onset effective therapy fly gene function human SNCAIP protein in vivo link protein loss of function mitochondrial dysfunction mutant neuron loss neuronal survival novel overexpression parkin gene/protein pars compacta prevent programs protein misfolding ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is a progressive neurodegenerative movement disorder. No treatment is currently available to prevent disease progression and neurodegeneration, although administration of L-dopa temporarily relieves parkinsonism. Understanding the molecular basis of PD is likely to facilitate development of effective therapies for the disease. We have recently found that proteins encoded by three recessive PD-linked genes, including parkin, PINK1 and DJ-1, form an novel E3 ligase complex to promote ubiquitination and degradation of the parkin substrates, parkin and synphilin-1. Pathogenic parkin and PINK1 mutants show impaired ability to degrade parkin and synphilin-1. Moreover, inactivation of Drosophila PINK1 (dPINKI) using RNAi results in loss of DA neurons and ommatidial degeneration, all of which are rescued by expression of human PINK1 (hPINKI). Furthermore, expression of human SOD1 suppresses neurodegeneration induced bydPINKI inactivation. Treatment of dPINKI RNAi flies with the antioxidants SOD and vitamin E significantly inhibits ommatidial degeneration. These studies demonstrate that dPINKI plays an essential role in maintaining neuronal survival, likely by antagonizing oxidative stress in neurons, and reveal a potential mechanism for neurodegeneration in PD caused by parkin, PINK1 or DJ-1 loss of function. We hypothesize that recessive PD-linked proteins parkin, PINK1 and DJ-1 function coordinately as an E3 ligase complex. Disease-associated mutations should impair E3 ligase activity, resulting in increased susceptibility of DA neurons to oxidative stress. To test this hypothesis, we will: 1. Determine serine/threonine kinase activity of PINK1 in the PPD E3 ligase complex. Hypothesis tested: PINKI's serine/threonine kinase activity plays important roles in regulating PPD E3 ligase activity via phosphorylating components of the PPD complex. 2. Determine reciprocal regulation between PPD E3 activity and oxidative stress. Hypothesis tested: PPD complex prevents cells from undergoing oxidative stress via its E3 ligase activity. Reduced PPD E3 activity leads to increased Oxidative stress. Oxidative stress reciprocally impair PPD E3 complex. 3. Define roles of the PPD complex in neuronal survival. Hypothesis tested: the PPD E3 ligase activity promotes neuronal survival via a mechanism of antagonizing oxidative stress.

描述（由申请人提供）：帕金森病（PD）是一种进行性神经退行性运动障碍。尽管服用左旋多巴可以暂时缓解帕金森病，但目前尚无治疗方法可以预防疾病进展和神经变性。了解帕金森病的分子基础可能有助于开发该疾病的有效疗法。我们最近发现，由三个隐性PD连锁基因（包括parkin、PINK1和DJ-1）编码的蛋白质形成一种新型E3连接酶复合物，以促进parkin底物parkin和synphilin-1的泛素化和降解。致病性 Parkin 和 PINK1 突变体降解 Parkin 和 synphilin-1 的能力受损。此外，使用 RNAi 灭活果蝇 PINK1 (dPINKI) 会导致 DA 神经元损失和小眼变性，所有这些都可以通过人 PINK1 (hPINKI) 的表达来挽救。此外，人 SOD1 的表达可抑制 dPINKI 失活引起的神经变性。用抗氧化剂 SOD 和维生素 E 处理 dPINKI RNAi 果蝇可显着抑制小眼变性。这些研究表明，dPINKI 在维持神经元存活中发挥着重要作用，可能是通过拮抗神经元中的氧化应激而发挥重要作用，并揭示了 Parkin、PINK1 或 DJ-1 功能丧失引起的 PD 神经变性的潜在机制。我们假设隐性 PD 连锁蛋白 Parkin、PINK1 和 DJ-1 作为 E3 连接酶复合物协调发挥作用。疾病相关突变会损害 E3 连接酶活性，导致 DA 神经元对氧化应激的敏感性增加。为了检验这一假设，我们将： 1. 确定 PPD E3 连接酶复合物中 PINK1 的丝氨酸/苏氨酸激酶活性。假设检验：PINKI 的丝氨酸/苏氨酸激酶活性通过磷酸化 PPD 复合物的成分在调节 PPD E3 连接酶活性中发挥重要作用。 2.确定PPD E3活性和氧化应激之间的相互调节。假设检验：PPD 复合物通过其 E3 连接酶活性防止细胞遭受氧化应激。 PPD E3 活性降低会导致氧化应激增加。氧化应激会相互损害 PPD E3 复合物。 3. 定义 PPD 复合物在神经元存活中的作用。假设检验：PPD E3 连接酶活性通过拮抗氧化应激的机制促进神经元存活。