Molecular Mechanism of the Cytoplasmic Dynein-Dynactin Motor Complex

细胞质动力蛋白-动力蛋白运动复合物的分子机制

• 批准号: 10366295
• 负责人: Arne Gennerich
• 金额: $ 37.97万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366295
• 关键词: ATP phosphohydrolase; Affect; Amino Acids; Behavior; Binding; Biological Assay; Brain; Cell physiology; Cerebral cortex; Child; Color; Complex; Congenital Abnormality; Data; Defect; Degenerative Disorder; Development; Disease; Dynein ATPase; Embryonic Development; Engineering; Epilepsy; Etiology; Fluorescence; Fluorescence Microscopy; Functional disorder; Future; Generations; Goals; Homologous Gene; Human; Infant; Intracellular Transport; Life; Link; Measurement; Mediating; Microtubules; Molecular; Molecular Conformation; Molecular Target; Motion; Motor; Movement; Mutagenesis; Mutation; Neurons; Output; Pattern; Process; Protein Engineering; Registries; Regulation; Reporting; Saccharomyces cerevisiae; Slide; System; Tail; Testing; Therapeutic Intervention; Time; Variant; base; brain malformation; cell motility; cofactor; developmental disease; dimer; dynactin; genetic regulatory protein; human disease; improved; innovation; insight; laser tweezer; lissencephaly; migration; nanoscale; nerve stem cell; nervous system disorder; novel; recruit; single molecule; stem cell division; therapy development

PROJECT SUMMARY/ABSTRACT Title: Molecular Mechanism of the Cytoplasmic Dynein-Dynactin Motor Complex Our long-term goal is to elucidate the molecular mechanism of the cytoplasmic dynein-dynactin motor complex, and to define the molecular bases of dynein-related diseases in humans. The microtubule motor cytoplasmic dynein, in cooperation with its regulatory proteins, drives several processes essential to embryonic development, including neuronal proliferation, neuronal migration, and the transport of numerous intracellular cargoes. Thus, it is not surprising dysfunction of dynein contributes to birth defects. A striking example is lissencephaly (LIS), a rare, but devastating brain malformation. Variants of this diseases are caused by loss or mutation of the dynein regulator Lis1. In lissencephaly, neurons fail to migrate properly during development and affected infants’ brains lack the normal gyral folding pattern and layering of the cerebral cortex. These children suffer untreatable epilepsy and severe psychomotor retardation, often dying in the first year of life. Unfortunately, mechanistic insights into dynein function and its regulation by Lis1 are limited, hampering our ability to understand the molecular basis for LIS, and other dynein-linked human diseases. In this proposal, we will combine single-molecule fluorescence and optical tweezers-based force measurements with innovative protein engineering to determine how cytoplasmic dynein in complex with its activator dynactin and the cargo adaptor bicaudal D (BicD), is regulated by Lis1. We will decipher how Lis1 regulates the motion and force generation of the dynein-dynactin-BicD (DDB) motor complex, and determine the effects of human disease mutations on Lis1-DDB function. These studies will provide a new understanding of dynein-linked diseases, and elucidate molecular targets for future therapeutic interventions.

项目摘要/摘要 标题：细胞质动力蛋白 - 二奈霉素运动复合物的分子机制 我们的长期目标是阐明细胞质动力蛋白 - 二奈霉素复合物的分子机制， 并定义人类动力蛋白相关疾病的分子碱基。微管运动细胞质 Dynein与其调节蛋白合作，驱动了胚胎必不可少的几个过程 发育，包括神经元增殖，神经元迁移和许多细胞内的运输 货物。这是毫不奇怪的是，动力蛋白的功能障碍会导致先天缺陷。罢工的例子是 lissphaly（LIS），一种罕见但毁灭性的脑畸形。这种疾病的变体是由损失或 动力蛋白调节剂LIS1的突变。在Lissencephaly，神经元在发育过程中无法正确迁移 受影响的婴儿的大脑缺乏正常的陀螺折叠模式和大脑皮层的分层。这些 儿童患有无法治疗的癫痫病和严重的心理运动迟缓，常常在生命的第一年死亡。 不幸的是，对动力蛋白功能的机理见解及其对LIS1的调节是有限的，这阻碍了我们 能够理解LIS的分子基础和其他与动力蛋白连接的人类疾病的能力。在这个建议中，我们 将结合单分子荧光和基于光学镊子的力量测量和创新 蛋白质工程以确定其激活剂Dynactin和货物的复合物中的细胞质动力蛋白如何 适配器Bicaudal D（BICD）由LIS1调节。我们将破译LIS1如何调节运动和力 生成动力蛋白 - 二奈霉素-BICD（DDB）运动复合物，并确定人类疾病的影响 LIS1-DDB功能的突变。这些研究将提供对动力蛋白连接疾病的新理解， 并阐明了未来治疗干预措施的分子靶标。