Biology of Parkin and It's Role in Parkinson's Disease

项目摘要-项目1：帕金氏病的生物学及其在帕金森病中的作用 帕金森病(PD)是一种复杂的神经退行性疾病，既有散发性的，也有家族性的。 Parkin基因突变是常染色体隐性遗传性帕金森病最常见的原因。在散发性帕金森病多巴胺能中， 氧化和亚硝化应激以及c-Abl的磷酸化导致parkin的抑制。因此，损失 Parkin功能是家族性和散发性帕金森病的基础。Parkin是一种E3连接酶，这种功能的丧失导致 底物AIMP2和PARIS的积累。我们发现AIMP2的表达会导致衰老 安乐死所致的依赖型DA神经退行性变。和巴黎的表达可能会导致丢失 促进神经退化的线粒体质量控制。 我们的假设是，散发性帕金森病的帕金森因亚硝化/氧化应激和c-Abl激活而失活 导致Y143(PY143 Parkin)上parkin的磷酸化和失活，随后积累 帕金底物，线粒体质量控制和毒性的损失。同时，α-突触核蛋白被磷酸化 在Y39(pY39α-突触核蛋白)上导致聚集和随后的毒性。自聚合α-突触核糖核酸以来 会导致线粒体功能障碍，这会造成一个前馈循环。 目的1：线粒体功能障碍是帕金森病的共同特征之一。这一目标将探索 Parkin基因突变引起的线粒体功能障碍与PARIS的关系我们已经证明了 PARIS是帕金森病的重要病理生理底物，在转录水平上抑制pGC-1αa 调控线粒体生物发生和线粒体氧化应激的主要转录共激活因子 回应。 目的2：Parkin失活会导致AIMP2和PARIS的积聚。AIMP2的表达 或者巴黎足以促进年龄依赖性DA神经退行性变。激活的事件顺序 PARIS和AIMP2将被探索以确定这两种蛋白质是否以及如何相互作用来启动细胞 死亡计划，死神。 目的：观察A53Tα-突触核蛋白转基因小鼠PY143Parkin和AIMP2、PARIS表达的变化。 提出了Parkin失活、PARIS和AIMP2上调以及PARP1激活是否起作用的问题 在α-突触核蛋白诱导的神经变性中起什么作用？这种可能性将通过α-突触核蛋白来探索 帕金森病预形成纤维(Pff)模型。 目标4：最先进的技术，包括深度测序和SILAM(氨基稳定同位素标记 哺乳动物中的酸)将被用于识别受成体条件性条件调控的基因和蛋白质 Parkin基因敲除及其与paris诱导的关系，目的是识别信号中的结点 神经退行性变的级联，可为帕金森病的治疗提供新的靶点。

PROJECT SUMMARY - PROJECT 1: BIOLOGY OF PARKIN AND ITS ROLE IN PARKINSON'S DISEASE Parkinson's disease (PD) is a complex neurodegenerative disorder that is both sporadic and familial. Mutations in parkin are the most common cause of autosomal recessive PD. In sporadic PD dopaminergic, oxidative and nitrosative stress as well as c-Abl phosphorylation result in inhibition of parkin. Thus, loss of parkin function is elemental to both familial and sporadic PD. Parkin is an E3 ligase, this loss of function leads to accumulation of the substrates, AIMP2 and PARIS. We have found that AIMP2 expression leads to age dependent DA neurodegeneration due to parthanatos. And PARIS expression may lead to loss of mitochondrial quality control that promotes neurodegeneration. Our hypothesis is that parkin inactivation in sporadic PD by nitrosative/oxidative stress, and c-Abl activation leads to phosphorylation of parkin on Y143 (pY143 parkin) and inactivation followed by the accumulation of parkin substrates, loss of mitochondrial quality control and toxicity. In parallel, α-synuclein is phosphorylated on Y39 (pY39 α-synuclein) resulting in aggregation and subsequent toxicity. Since aggregated α-synuclein can lead to mitochondrial dysfunction it creates a feed forward cycle. Aim 1: One of the unifying features of PD is mitochondrial dysfunction. This aim will explore the inter- relationship of PARIS and mitochondrial dysfunction caused by mutations in parkin. We have shown that PARIS is an important pathophysiologic substrate of parkin in PD that transcriptionally represses PGC-1α a major transcriptional co-activator that regulates mitochondrial biogenesis and mitochondrial oxidant stress responses. Aim 2: Inactivation of parkin results in accumulation of both AIMP2 and PARIS. Expression of either AIMP2 or PARIS is sufficient to promote age dependent DA neurodegeneration. The sequence of events activated by PARIS and AIMP2 will be explored to determine if and how these two proteins interact to initiate the cell death program, parthanatos. Aim 3: We observe pY143 parkin and elevated AIMP2 and PARIS in A53T α-synuclein transgenic mice that raises the question of whether parkin inactivation, PARIS and AIMP2 upregulation and PARP1 activation play a role in α-synuclein induced neurodegeneration? This possibility will be explored with the α-synuclein preformed fibrils (PFFs) model of PD. Aim 4: State-of-the-art technology including deep sequencing and SILAM (stable isotope labeling by amino acids in mammals) will be deployed to identify genes and proteins that are regulated by adult conditional knockout of parkin and their relationship to PARIS induction with the goal of identify nodal points in the signal cascade of neurodegeneration that can provide new targets for the treatment of PD.