THE ROLE OF NUB1 IN PATHOGENESIS OF PARKINSON'S DISEASE

NUB1 在帕金森病发病机制中的作用

• 批准号: 7090310
• 负责人: TETSU KAMITANI
• 金额: $ 24万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-15 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7090310
• 关键词: Lewy body; Parkinson' s disease; RNA interference; gene expression; gene mutation; gene targeting; genetically modified animals; immunoprecipitation; laboratory mouse; nerve /myelin protein; parkin gene /protein; pathologic process; proteasome; protein protein interaction; protein structure function; transfection; ubiquitin; western blottings

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the loss of dopaminergic neurons and the presence of cytosolic inclusions, called Lewy bodies (IBs). Mutations in the gene encoding alpha-synuclein, parkin, or synphilin-1 have been reported in patients with familial or sporadic Parkinson's disease. Recently, both alpha-synuclein and synphilin-1 were shown to be ubiquitinated through the E3 ubiquitin ligase activity of wild-type parkin, but not by familial-linked mutant parkin. Moreover, the coexpression of alpha-synuclein, synphilin-1, and parkin resulted in the formation of LB-like ubiquitin-positive cytosolic inclusions in cultured cells. Importantly, familial-linked mutations in parkin disrupted the formation of the ubiquitin-positive inclusions. These observations indicate that defects in the ubiquitination of LB-associated proteins are involved in the pathogenesis of Parkinson's disease. NEDD8 is a ubiquitin-like protein that covalently modifies target proteins in a manner analogous to ubiquitination. Recently, we identified a novel NEDD8-interacting protein, NUB1, that functioned like a molecular chaperone to recruit NEDD8 to the S5a subunit of the 26S proteasome for degradation. In addition to NEDD8, NUB1 interacted with synphilin-1 through its NEDDS-binding site, implying that NUB1 also recruits synphilin-1 to the S5a subunit of the 26S proteasome. Because parkin binds to the same subunit, we hypothesize that NUB1 recruits synphilin-1 to the S5a subunit for its ubiquitination by parkin. Furthermore, we hypothesize that the targeting of synphilin-1 to the S5a subunit is involved not only in the ubiquitination of synphilin-1, but also in LB formation. Indeed, we detected NUB1 in the LBs of brains from patients with Parkinson's disease, suggesting that NUB1 plays a role in LB formation or its breakdown. To test these hypotheses, we will pursue 3 Aims, which are to define 1) the role of NUB1 in the targeting of synphilin-1 to the S5a subunit, 2) the role of NUB1 in the ubiquitination of synphilin-1, and 3) the role of NUB1 in the formation of cytosolic inclusions by synphilin-1. The outcome of our research proposed here will provide a molecular basis for the involvement of NUB1 in the pathogenesis of Parkinson's disease. Furthermore, the outcome will potentially lead to the development of new diagnostic methods and therapeutic targets for Parkinson's disease.

描述(申请人提供)：帕金森病(PD)是一种常见的神经退行性疾病，其特征是多巴胺能神经元丢失和胞浆内包涵体的存在，称为路易小体(IBS)。有报道称，家族性或散发性帕金森氏症患者的α-突触核蛋白、parkin或突触蛋白-1基因发生突变。最近，α-突触核蛋白和突触素-1都被证明是通过野生型parkin的E3泛素连接酶活性泛素化的，但不是通过家族性连锁突变的parkin。此外，α-突触核蛋白、突触素-1和parkin的共同表达导致培养细胞中形成类LB型泛素阳性胞浆内含物。重要的是，Parkin中的家族相关突变破坏了泛素阳性包涵体的形成。这些观察结果表明，LB相关蛋白泛素化的缺陷参与了帕金森病的发病机制。NEDD8是一种泛素样蛋白，以类似泛素化的方式共价修饰靶蛋白。最近，我们发现了一种新的NEDD8相互作用蛋白NUB1，它的功能类似于分子伴侣，将NEDD8招募到26S蛋白酶体的S5a亚单位进行降解。除了NEDD8外，NUB1还通过其NEDDS结合位点与SynPhilin-1相互作用，这意味着NUB1还将SynPhilin-1招募到26S蛋白酶体的S5a亚单位。由于Parkin与同一亚基结合，我们假设NUB1通过Parkin将SynPhilin-1招募到S5a亚基，使其泛素化。此外，我们假设SynPhilin-1靶向S5a亚基不仅参与了SynPhilin-1的泛素化，而且还参与了Lb的形成。事实上，我们在帕金森氏病患者的LBS大脑中检测到了NUB1，这表明NUB1在LB的形成或分解中发挥了作用。为了验证这些假说，我们将追求三个目标，这三个目标是定义1)NUB1在将SynPhilin-1靶向S5a亚基中的作用，2)NUB1在SynPhilin-1泛素化中的作用，以及3)NUB1在SynPhilin-1形成胞质内含物中的作用。我们在这里提出的研究结果将为NUB1参与帕金森病的发病机制提供分子基础。此外，这一结果可能会导致帕金森病新的诊断方法和治疗目标的开发。