Mechanisms of Lis1 and NudE/L Regulation of the Molecular Motor Dynein

Lis1 和 NudE/L 分子运动动力蛋白的调节机制

• 批准号: 10087571
• 负责人: Morgan DeSantis
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10087571
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Adaptor Signaling Protein; Address; Affect; Affinity; Award; Binding; Biochemistry; Biotinylation; Cell Cycle; Cell division; Cell physiology; Cells; Cellular biology; Copy Number Polymorphism; Cryoelectron Microscopy; Data; Data Set; Disease; Dynein ATPase; Genes; Goals; Health; Homologous Gene; Human; Impairment; In Situ; In Vitro; Interphase; Knowledge; Learning; Mechanics; Mentors; Microcephaly; Microscopy; Microtubules; Mitosis; Mitotic; Molecular Motors; Motor; Movement; Mutation; Neurodegenerative Disorders; Neurons; Pathway interactions; Phase; Phosphorylation; Play; Plus End of the Microtubule; Post-Translational Protein Processing; Process; Proteins; Proteomics; Recombinant Proteins; Regulation; Regulatory Pathway; Research; Role; Schizophrenia; Structural Protein; Structure; Techniques; Testing; Training; United States National Institutes of Health; Walking; Work; autism spectrum disorder; base; cell motility; cell type; developmental disease; experimental study; genetic regulatory protein; human disease; in vitro activity; interdisciplinary approach; lissencephaly; live cell imaging; migration; novel; protein complex; protein reconstitution; structural biology; trafficking

PROJECT SUMMARY/ABSTRACT Microtubule-based transport is required for cell division, cell migration, and for transport of a number of cellular cargoes. A number of neurodevelopmental and neurodegenerative diseases are caused by or associated with impaired microtubule-based transport. Cytoplasmic dynein 1 (dynein) is one of two molecular motor proteins that are responsible for microtubule-based transport. Dynein is a highly regulated motor and interacts with a number of adaptor proteins that modulate its function and activity. Mutations or copy number variations of dynein regulatory proteins also leads to neurodevelopmental diseases. Despite the importance to human health, mechanisms of how dynein is regulated are largely unknown. This proposal for an NIH K99/R00 Pathway to Independence Award seeks to understand how Lis1 and NudE/L, which are two regulators required for nearly every dynein function, modulate dynein activity. Impaired Lis1 and NudE/L function is implicated in a number of human diseases, including microcephaly, lissencephaly, schizophrenia, and autism. In Aim1 during the mentored phase of the award, Dr. DeSantis will determine how Lis1 and NudE/L alter dynein function using a combination of structural biology and pure protein reconstitution experiments. Phosphorylation of dynein, Lis1, and NudE/L alter their activity but the mechanism of how this occurs in unknown. During the independent phase of the award, Dr. DeSantis will also determine how post-translational modifications influence dynein, Lis1, and NudE/L activity. In Aim 2, Dr. DeSantis will identify novel dynein regulatory pathways using a combination of proteomics, cell biology, live cell imaging, and recombinant protein reconstitutions. Dr. DeSantis has already identified novel Lis1 and NudE/L interacting proteins and will elucidate their function and mechanism during the mentored and independent award phase. The results of this work will reveal mechanisms of dynein regulation, which has far reaching implications in human health and disease. Dr. DeSantis will receive training in cryo-electron microscopy in the K99 portion of the award. When combined with her background in biochemistry, cell biology, and live cell imaging, learning cryo-electron microscopy will empower Dr. DeSantis' research about mechanisms of motor protein regulation far beyond the duration of the K99/R00 award.

项目摘要/摘要 基于微管的运输是细胞分裂，细胞迁移和许多细胞运输所必需的 货物。许多神经发育和神经退行性疾病是由或与 基于微管的运输受损。细胞质动力蛋白1（动力蛋白）是两种分子运动蛋白之一 负责基于微管的运输。 Dynein是一种高度调节的电动机，与A相互作用 调节其功能和活性的衔接蛋白的数量。突变或拷贝数变化 动力蛋白调节蛋白也会导致神经发育疾病。尽管对人类很重要 健康，如何调节动力蛋白的机制在很大程度上尚不清楚。该NIH K99/R00的建议 独立之途径旨在了解LIS1和NUDE/L是如何需要的两个监管机构的 对于几乎每个动力蛋白功能，可调节动力蛋白活性。 LIS1和裸/L功能受损与A有关 人类疾病的数量，包括小头畸形，Lissencephaly，精神分裂症和自闭症。在AIM1期间 奖项的指导阶段DeSantis博士将确定Lis1和Nude/L使用Dynein功能如何使用 结构生物学和纯蛋白质重构实验的结合。动力蛋白的磷酸化， lis1和裸体/l改变了其活性，但在未知中发生这种情况的机制。在独立期间 奖项的阶段，DeSantis博士还将确定翻译后修饰如何影响Dynein， Lis1和裸性/L活性。在AIM 2中，Desantis博士将使用A的新型动力蛋白调节途径 蛋白质组学，细胞生物学，活细胞成像和重组蛋白质重组的结合。 DeSantis博士 已经确定了新颖的Lis1和裸/L相互作用蛋白，并将阐明其功能， 指导和独立奖励阶段的机制。这项工作的结果将揭示 动力蛋白调节的机制，对人类健康和疾病具有很大的影响。博士 DeSantis将在该奖项的K99部分中接受冷冻电子显微镜的培训。合并时 她在生物化学，细胞生物学和活细胞成像方面的背景，学习冷冻电子显微镜将 增强DeSantis博士对运动蛋白调节机制的研究，远远超出了持续时间 K99/R00奖。