Mechanism of Action of Dynactin

Dynactin 的作用机制

• 批准号: 8548381
• 负责人: Richard Bert Vallee
• 金额: $ 40.97万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-28 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8548381
• 关键词: Acute; Axonal Transport; Binding; Biological Assay; Brain; Cell division; Cell physiology; Cells; Chromosome Segregation; Complex; Defect; Development; Disease; Dynein ATPase; Excision; Generations; Genetic; Human; In Vitro; Individual; Intracellular Transport; Life; Light; Methods; Microtubules; Motor; Motor Activity; Mutation; Nerve Degeneration; Organelles; Physiological; Post-Translational Protein Processing; Production; Protein Isoforms; Proteins; RNA Interference; Recombinants; Regulation; Relative (related person); Reporting; Resolution; Role; Sensory; Site; Structure; System; Tail; Testing; Text; Ursidae Family; Vesicle; Work; abstracting; base; cell behavior; cell motility; cofactor; dynactin; in vitro Assay; in vitro testing; in vivo; motor neuron degeneration; neoplastic; novel; particle; single molecule

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Cytoplasmic dynein is a microtubule motor protein involved in a very wide array of cellular functions, including vesicular transport, chromosome segregation, and cell migration. A single major form of cytoplasmic dynein is responsible for almost all aspects of these activities, but how it is adapted to such a diversity of functions at a broad range of subcellular sites remains a major question. Among the known dynein interactors, two complexes have emerged with prominent roles in dynein cargo binding and motor regulation, NudE-LIS1 and dynactin. We have recently reported NudE-LIS1 to have a novel and unique ability to interact with the dynein motor domain during its powerstroke and adapt the motor protein for high load functions. We also found LIS1-NudE to compete with dynactin for dynein binding, suggesting that the two complexes may serve in alternative regulatory roles. This proposal is to test this hypothesis and to bring to bear on dynactin the approaches we have successfully used to determine the functions and mechanism of action of LIS1 and NudE. Dynactin is known to enhance dynein processivity in vitro. However, its interaction with dynein has been difficult to control, hampering progress toward a complete understanding of its mechanochemical functions. Because dynactin is also important in dynein cargo recruitment, its specific in vivo role in motor regulation has also been difficult to define. Preliminary results have identified conditions controlling the dynein-dynactin interaction, and have revealed potent, long-range allosteric effects for dynactin fragments on dynein force production and processivity. The Aims of this proposal are (1) to determine how the dynein-dynactin interaction is regulated and to produce and define cocomplexes for further analysis; (2) to determine the complete scope of regulatory functions for the dynactin complex, its major regulatory subunit p150Glued, and its subfragments to understand the underlying mechanisms for dynein regulation; and (3) to determine the specific roles of dynactin in dynein motor regulation in vivo by high resolution particle tracking and force analysis. These studies are of broad relevance for understanding basic mechanisms of cell behavior. In addition, they should shed important new light into the mechanisms underlying brain developmental disease, motor neuron degeneration, cell division, and other physiological and pathophysiological functions.

在此输入文本，它是您的应用程序的新摘要信息。此部分必须为no 超过30行的文本。 细胞质动力蛋白是一种微管运动蛋白，参与多种细胞 功能，包括囊泡运输、染色体分离和细胞迁移。一个 细胞质动力蛋白的单一主要形式几乎负责这些方面的所有方面 活动，而是如何在广泛的亚细胞范围适应如此多样化的功能 选址仍然是一个主要问题。在已知的动力蛋白相互作用分子中，有两个复合体 在动力蛋白结合和运动调节中发挥重要作用，裸体-LIS1和 动力蛋白。我们最近报道裸体-LIS1具有一种新颖而独特的相互作用能力 在强力性卒中时与动力蛋白运动区结合，并使运动蛋白适应高负荷 功能。我们还发现了Lis1-nude与dynactin竞争动力蛋白结合，这表明 这两个联合体可能会扮演不同的监管角色。这项提议是为了测试这一点 并将我们成功使用的方法应用到dynactin上 确定Lis1和nude的功能和作用机制。已知的动力蛋白 增强动力蛋白的体外加工能力。然而，它与动力蛋白的相互作用一直很难 控制，阻碍了对其机械力化学的完全了解的进展 功能。因为动力蛋白在动力蛋白货物募集中也很重要，所以它在体内的特异性 在运动调节中的作用也很难定义。初步结果显示， 控制动力蛋白-动力蛋白相互作用的条件，并揭示了强大的、远程的 动力蛋白片段对动力蛋白作用力产生和加工的变构效应。这个 这项建议的目的是(1)确定动力蛋白-动力蛋白相互作用是如何调节的 并产生和定义用于进一步分析的络合物；(2)确定完整的 其主要调控亚单位--动力蛋白复合体的调控功能范围 P150Glued及其亚片段，以了解动力蛋白的潜在机制 以及(3)确定dynactin在动力蛋白运动调节中的具体作用。 VIVO通过高分辨率粒子跟踪和力分析。这些研究是广泛的。 与理解细胞行为的基本机制有关。此外，他们应该摆脱 对大脑发育疾病、运动的潜在机制的重要新认识 神经元变性、细胞分裂等生理和病理生理功能。