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之相关的 基于微管的运输受损。细胞质动力蛋白1(Dynein)是两种分子马达蛋白之一 负责基于微管的运输。动力蛋白是一种高度调节的马达，它与一种 调节其功能和活性的接头蛋白的数量。基因突变或拷贝数变异 动力蛋白调节蛋白也会导致神经发育疾病。尽管对人类来说很重要 在健康方面，动力蛋白如何调节的机制在很大程度上是未知的。NIH K99/R00的此建议书 独立之路奖旨在理解LIS1和裸体/L这两个监管机构如何被要求 对于几乎每一种动力蛋白功能，都要调节动力蛋白的活动。LIS1和裸体/L功能受损涉及 许多人类疾病，包括小头症、无脑畸形、精神分裂症和自闭症。在Aim1期间 在该奖项的指导阶段，德桑蒂斯博士将确定LIS1和裸体/L如何使用 结构生物学和纯蛋白质重组实验的结合。动力蛋白的磷酸化， 和裸体/L改变了他们的活动，但这是如何发生的机制尚不清楚。在独立期间 在颁奖阶段，德桑蒂斯博士还将确定翻译后修饰如何影响动力蛋白， LIS1，和裸体/L活动。在Aim 2中，DeSantis博士将使用一种新的动力蛋白调节通路 蛋白质组学、细胞生物学、活细胞成像和重组蛋白重组的结合。德桑蒂斯博士 已经发现了新的Lis1和nude/L相互作用蛋白，并将阐明它们的功能和 在指导和独立颁奖阶段的机制。这项工作的结果将揭示 动力蛋白的调节机制，对人类健康和疾病具有深远的影响。Dr。 DeSantis将在K99奖项部分接受冷冻电子显微镜方面的培训。当与 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 DeSantis博士对运动蛋白调节机制的研究远远超出了 K99/R00奖。