Mechanism of Action of Dynactin

Dynactin 的作用机制

• 批准号: 8343218
• 负责人: Richard Bert Vallee
• 金额: $ 44.15万
• 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/8343218
• 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; Ursidae Family; Vesicle; Work; 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.

在此处输入文本，该文本是您的应用程序的新摘要信息。此部分必须没有 文本长度超过 30 行。 细胞质动力蛋白是一种微管运动蛋白，参与非常广泛的细胞活动 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负责这些的几乎所有方面 活动，但它如何适应广泛的亚细胞的如此多样化的功能 网站仍然是一个主要问题。在已知的动力蛋白相互作用子中，有两种复合物 在动力蛋白货物结合和运动调节、NudE-LIS1 和 动力蛋白。我们最近报道 NudE-LIS1 具有新颖且独特的交互能力 在动力冲程期间与动力蛋白运动结构域结合，并使运动蛋白适应高负载 功能。我们还发现 LIS1-NudE 与 dynactin 竞争动力蛋白结合，这表明 这两个复合体可以发挥替代的监管作用。这个提议就是为了测试这个 假设并将我们已成功使用的方法应用于动力蛋白 确定 LIS1 和 NudE 的功能和作用机制。 Dynactin 已知 增强动力蛋白的体外合成能力。然而，其与动力蛋白的相互作用一直难以确定。 控制，阻碍了对其机械化学的全面理解的进展 功能。由于动力蛋白在动力蛋白货物招募中也很重要，因此它在体内具有特异性 在运动调节中的作用也很难定义。初步结果已确定 控制动力蛋白-动力蛋白相互作用的条件，并揭示了有效的、长程的 动力蛋白片段对动力蛋白力产生和持续合成能力的变构效应。这 该提案的目的是 (1) 确定如何调节动力蛋白-动力蛋白相互作用 并产生和定义复合物以供进一步分析； (2)确定完整 动力蛋白复合体（其主要调节亚基）的调节功能范围 p150Glued 及其子片段，用于了解动力蛋白的基本机制 规定; (3) 确定 dynactin 在动力蛋白运动调节中的具体作用 体内通过高分辨率粒子跟踪和力分析。这些研究具有广泛的 与理解细胞行为的基本机制的相关性。此外，他们应该脱掉 为大脑发育疾病、运动疾病的潜在机制提供了重要的新线索 神经元变性、细胞分裂以及其他生理和病理生理功能。