Structure and function of DISC1 in the cAMP pathway

cAMP 通路中 DISC1 的结构和功能

• 批准号: 10470407
• 负责人: Julien Roche
• 金额: $ 32.41万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470407
• 关键词: Affinity; Antidepressive Agents; Binding; Binding Proteins; Biochemical; Biological Assay; Bipolar Depression; Bipolar Disorder; C-terminal; Complex; Computational Technique; Computer Simulation; Cyclic AMP; DISC1 gene; DNA; DNA Sequence; Data; Deuterium; Development; Disease; Elements; Enzymes; Etiology; Family; Foundations; Functional disorder; Genetic Transcription; Goals; Hydrogen; In Vitro; Individual; Length; Major Depressive Disorder; Mass Spectrum Analysis; Memory; Mental disorders; Molecular; Mutation; N-terminal; NMR Spectroscopy; Neuraxis; Neurodegenerative Disorders; Neurons; Nuclear; PDE4B; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Predisposing Factor; Protein Isoforms; Proteins; Regulation; Reporting; Research; Research Project Grants; Roentgen Rays; Role; Scaffolding Protein; Schizophrenia; Shapes; Signal Pathway; Structural Models; Structure; Techniques; Testing; Therapeutic; Ulysses Contracts; Variant; Work; X-Ray Crystallography; base; dimer; drug discovery; experimental study; genetic corepressor; genetic risk factor; genomic locus; inhibitor; insight; member; molecular dynamics; nervous system disorder; neurotransmission; novel therapeutic intervention; phosphoric diester hydrolase; reconstitution; simulation; transcription factor

DISC1 (Disrupted-In-Schizophrenia-1) has emerged as a significant genetic risk factor for a wide range of mental illness such as schizophrenia, bipolar disorders and depression. The product of DISC1 is a long scaffold protein that plays a critical role in several neuronal signaling pathways. Despite growing appreciation of its role in the etiology of mental disorders, almost nothing is known about the structure of DISC1 and its mechanisms of interaction and regulation. This research project will specifically focus on the role of DISC1 in the cAMP pathway. Reduced cAMP levels are found in patients suffering from major depression and are associated with neurodegenerative diseases. Enzymes controlling intracellular levels of cAMP, such as the phosphodiesterase family PDE4, have considerable pharmaceutical importance for the development of antidepressant and memory enhancing drugs. The overall goal of this project is to provide a structural framework describing the role of DISC1 in the cAMP pathway and to understand how alterations in this pathway contribute to the etiology of psychiatric disorders. Aim 1 will provide a complete structural model of DISC1, including its mechanism of oligomerization and modulation by phosphorylation. We will also investigate the structural consequences of disease-associated mutations. We are proposing here an integrative structural approach combining Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS), solution NMR spectroscopy, X-ray crystallography, Small-angle X-ray Scattering (SAXS) and molecular simulations to reconstitute the complete structural and dynamical features of DISC1. Our preliminary data demonstrate that we can characterize DISC1 N-terminal constructs by solution NMR spectroscopy while constructs encompassing the central and/or C-terminal domains are well suited for X- ray crystallography study. Aim 2 will combine biochemical and structural approaches to characterize the interaction between DISC1 and ATF4, a major transcription factor the cAMP pathway controlling the expression of the CRE-elements and of the phosphodiesterase PDE4D9. Our preliminary data show that DISC1-ATF4 complex can be reconstituted in vitro with truncated constructs of both proteins. This aim will provide a mechanistic framework for understanding the role of DISC1 as transcriptional co-repressor and the function of DISC1-ATF4 complex in the cAMP pathway. Aim 3 will unravel the mechanisms of allosteric inhibition of long and short PDE4 isoforms by DISC1, using a combination of biochemical, structural and computational techniques. Overall, completion of this project will provide a comprehensive, unifying framework for understanding the role of DISC1 in the cAMP pathway. Our work will also provide new structural targets for the development of molecules regulating cAMP levels.

DISC1（Disrupted-In-Schizophrenia-1）已成为多种疾病的重要遗传风险因素。 一系列精神疾病，如精神分裂症、双相情感障碍和抑郁症。 DISC1的产品是 一种长支架蛋白，在多种神经元信号传导通路中发挥关键作用。尽管成长 人们对其在精神障碍病因学中的作用的认识，几乎对它的结构一无所知。 DISC1 及其相互作用和调节机制。该研究项目将特别关注 DISC1 在 cAMP 通路中的作用。患有严重疾病的患者中发现 cAMP 水平降低 抑郁症与神经退行性疾病有关。控制细胞内水平的酶 cAMP，例如磷酸二酯酶家族 PDE4，对于以下疾病具有相当大的药学重要性： 抗抑郁药和记忆增强药物的开发。该项目的总体目标是 提供一个结构框架来描述 DISC1 在 cAMP 通路中的作用并了解如何 该途径的改变导致精神疾病的病因学。目标 1 将提供完整的 DISC1 的结构模型，包括其寡聚化和磷酸化调节机制。 我们还将研究与疾病相关的突变的结构后果。我们提议 这是结合氢-氘交换质谱的综合结构方法 (HDX-MS)、溶液核磁共振波谱、X 射线晶体学、小角 X 射线散射 (SAXS) 和 分子模拟来重建 DISC1 的完整结构和动力学特征。我们的 初步数据表明我们可以通过溶液 NMR 表征 DISC1 N 末端结构 光谱，而包含中心和/或 C 末端结构域的构建体非常适合 X- 射线晶体学研究。目标 2 将结合生化和结构方法来表征 DISC1 和 ATF4 之间的相互作用，ATF4 是 cAMP 通路中控制 CRE 元件和磷酸二酯酶 PDE4D9 的表达。我们的初步数据表明 DISC1-ATF4 复合物可以在体外用两种蛋白质的截短结构重建。这一目标将 为理解 DISC1 作为转录辅阻遏物的作用提供了一个机制框架 DISC1-ATF4 复合物在 cAMP 通路中的功能。目标 3 将揭示变构机制 DISC1 结合使用生化、结构和抑制长和短 PDE4 亚型 计算技术。总体而言，该项目的完成将提供一个全面、统一的 理解 DISC1 在 cAMP 通路中的作用的框架。我们的工作也将提供新的 开发调节 cAMP 水平的分子的结构目标。