Understanding the mechanisms of UBE3A regulation in neuronal development

了解 UBE3A 在神经元发育中的调节机制

• 批准号: 10531212
• 负责人: Jason J Yi
• 金额: $ 39.38万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10531212
• 关键词: Affect; Angelman Syndrome; Animals; Benign; Binding; Binding Proteins; Binding Sites; Biochemical; Biological Assay; Bipolar Disorder; Calorimetry; Catalogs; Cells; Cryoelectron Microscopy; Disease; Dose; Engineering; Enzymes; Genes; Goals; Hyperactivity; Individual; Kinetics; Length; Link; Literature; Measures; Mental Depression; Mental disorders; Methods; Modeling; Mood Disorders; Mus; Mutation; Neurons; Pathology; Phenotype; Protein Engineering; Proteins; Regulation; Regulatory Element; Reporting; Research; Schizophrenia; Site; Structure; Synapses; Testing; Therapeutic Intervention; Titrations; UBE3A gene; Ubiquitin; Variant; WNT Signaling Pathway; Work; chromosome 15q duplication syndrome; developmental disease; enzyme mechanism; experimental study; gain of function; gain of function mutation; high throughput screening; in vivo; insight; interest; loss of function; loss of function mutation; neuron development; neuropsychiatric disorder; neuropsychiatry; novel; public health relevance; tool; ubiquitin ligase; ubiquitin-protein ligase

Abstract. UBE3A is a gene that encodes a HECT (Homologous to E6AP C-terminus) domain E3 ubiquitin ligase linked to numerous developmental and psychiatric disorders. Loss of function of the maternally derived UBE3A enzyme causes Angelman syndrome whereas gain of function, due to duplication or triplication of UBE3A, is linked to a broad range of disorders including Dup15q syndrome, schizophrenia, and mood disorders. These observations strongly suggest that bi-directional changes in UBE3A activity contribute to neuropsychiatric pathology. However, the mechanisms that regulate UBE3A activity remain poorly understood. The primary goal of our proposed research is to uncover mechanisms of UBE3A regulation, that when faulty, can lead to aberrant gain or loss of UBE3A function. In preliminary work, we developed a high-throughput assay to assess the functional consequence of non-truncating UBE3A missense variants. This screen identified numerous novel loss of function mutations, as well as gain of function mutations that hyperactivate UBE3A activity well above wild type (WT) enzyme levels. These results provide deep structure-function information that we can now leverage to uncover mechanisms of UBE3A regulation. This proposal aims to, 1) create a complete functional catalogue of known missense variants identified in individuals, 2) utilize structure-function analyses to identify mechanisms that can lead to both aberrant gain and loss of enzyme function, and 3) leverage biochemical insights to engineer proteins that can target UBE3A activity. The molecules generated from our work will be applied to examine whether alteration of UBE3A activity can rescue synaptic phenotypes observed in mice harboring hyperactivating mutations in UBE3A. If successful, our work will provide new biochemical insights and tools that will make it possible to target UBE3A for therapeutic intervention in various neuropsychiatric disorders.

抽象的。UBE3A是一个编码Hect(与E6AP C末端同源)结构域E3泛素连接酶的基因 与许多发育和精神障碍有关。母源性UBE3A功能丧失 酶导致Angelman综合征，而由于UBE3A的复制或三倍而导致的功能获得是 与包括Dup15q综合征、精神分裂症和情绪障碍在内的一系列疾病有关。这些 观察有力地表明，UBE3A活动的双向变化有助于神经精神病学 病理学。然而，调控UBE3A活动的机制仍然知之甚少。首要目标 我们提出的研究的目的是揭示UBE3A调控的机制，当错误时，可能会导致异常 UBE3A功能的得失。在前期工作中，我们开发了一种高通量检测方法来评估 非截断UBE3A错义变体的功能后果。这个屏幕识别出了无数的小说 功能突变的丧失，以及远高于UBE3A活性的功能突变的获得 野生型(WT)酶水平。这些结果提供了我们现在所能提供的深层结构-功能信息 利用杠杆揭示UBE3A监管机制。本提案旨在，1)创建一个完整的功能 在个体中识别的已知错义变体的目录，2)利用结构-功能分析来识别 可导致酶功能异常获得和丧失的机制，以及3)利用生化 对能够针对UBE3A活性的工程蛋白质的见解。我们的工作产生的分子将是 应用于检测UBE3A活性改变是否能挽救小鼠观察到的突触表型 在UBE3A中存在过度激活突变。如果成功，我们的工作将提供新的生化见解和 将有可能针对UBE3A进行各种神经精神障碍的治疗干预的工具。