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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要细胞分裂、细胞迁移和许多细胞的运输需要基于微管的运输货物。许多神经发育和神经退行性疾病是由以下疾病引起或与之相关：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。动力蛋白是一种高度调节的马达，并与调节其功能和活性的接头蛋白的数量。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。尽管对人类很重要健康方面，动力蛋白的调节机制在很大程度上尚不清楚。本提案针对 NIH K99/R00 独立之路奖旨在了解 Lis1 和 NudE/L（这两个监管机构）如何需要对于几乎所有动力蛋白功能，调节动力蛋白活性。 Lis1 和 NudE/L 功能受损与许多人类疾病，包括小头畸形、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，DeSantis 博士将确定 Lis1 和 NudE/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， Lis1 和 NudE/L 会改变它们的活性，但其发生机制尚不清楚。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 NudE/L 活性。在目标 2 中，DeSantis 博士将使用蛋白质组学、细胞生物学、活细胞成像和重组蛋白重建的结合。德桑蒂斯博士已经鉴定出新的 Lis1 和 NudE/L 相互作用蛋白，并将阐明它们的功能和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机制，对人类健康和疾病具有深远影响。博士。 DeSantis 将在该奖项的 K99 部分接受冷冻电子显微镜培训。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使 DeSantis 博士关于运动蛋白调节机制的研究远远超出了持续时间 K99/R00奖。