Coordination of molecular motor activity in intracellular transport and assembly of cytoskeletal architecture.

细胞内运输和细胞骨架结构组装中分子运动活动的协调。

• 批准号: 9382131
• 负责人: Richard James McKenney
• 金额: $ 37.6万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382131
• 关键词: ATP Hydrolysis; Actins; Architecture; Behavior; Biochemistry; Biological; Biological Assay; Cell physiology; Cells; Chemicals; Cilia; Complex; Disease; Dynein ATPase; Endocytic Vesicle; Environment; Eukaryota; Filament; Goals; Health; Homeostasis; Huntington Disease; Impairment; In Vitro; Individual; Intracellular Transport; Kinesin; Link; Malignant Neoplasms; Microtubules; Mitochondria; Mitotic spindle; Molecular; Molecular Machines; Molecular Motors; Motion; Motor; Motor Activity; Motor output; Pathologic; Process; Property; Proteins; Recruitment Activity; Regulation; Research; Structure; System; Testing; Translating; Work; cell motility; in vivo; insight; novel; rab GTP-Binding Proteins; reconstitution; self assembly; single molecule; spatiotemporal; tool

Title: Coordination of molecular motor activity in intracellular transport and assembly of cytoskeletal architecture. P.I. – Richard J. McKenney Research Summary Intracellular transport is essential for cellular homeostasis in eukaryotes. Much of this process is carried out by molecular motors that convert the chemical energy from ATP hydrolysis into motion along the actin and microtubule cytoskeletal networks. Decades of research has uncovered structural and molecular details that explain how many of these motors move along their filament tracks in isolation. In the cellular milieu, most of these motors act in concert with complex regulatory machinery that links them to their respective cargos, modulates their motile properties, and dictates spatiotemporal activity. How individual motor output is controlled by this machinery is currently not clear and difficult to dissect in the complex environment of the cell. In addition, many cargos are moved simultaneously by motors of opposite polarity, in a process called bidirectional transport. How individual motors are recruited to cargo, activated, and integrated with other classes of motors presents a large challenge to the field. Further, the activities of disparate motors are harnessed to build and maintain critical cytoskeletal structures such as the mitotic spindle, cilium, and cleavage furrow. How motor and regulatory activities are coordinated to drive the self-assembly of such structures is currently a significant barrier to understanding normal and diseased cellular physiology. This application seeks to develop novel assays and tools to study the complexity of motor recruitment and regulation, bidirectional transport of cargos, and the self-assembly of cytoskeletal structures driven by motors and associated molecules. Our approach to combine biochemistry and single- molecule analysis towards in vitro reconstitutions that test molecular function, and translate our findings into in vivo systems that test hypotheses generated by these reconstitutions, will open up fruitful long-term avenues of research. We propose to: 1) Reconstitute and study the recruitment, regulation, and motility of cytoplasmic dynein and kinesin motors bound to membranous cargo through the endogenous Rab GTPase machinery that is known to link these motors to endocytic vesicles and mitochondria in cells, and 2) Reconstitute and study functions of dynein and kinesin motors that drive the self-assembly of the mitotic spindle. These broad goals build and expand on our expertise and previous work in dissecting the regulatory mechanisms of the cytoplasmic dynein motor, and aim to provide powerful new tools useful towards dissecting complex motor function. Our work will illuminate basic molecular and cell biological principles that drive cellular homeostasis and provide insight into the pathological mechanisms that arise from molecular motor malfunction.

标题：细胞内运输和细胞骨架组装中分子运动活动的协调 建筑学。 P.I.——理查德·J·麦肯尼 研究总结 细胞内运输对于真核生物的细胞稳态至关重要。 由分子马达执行，将 ATP 水解的化学能转化为运动 数十年的研究揭示了肌动蛋白和微管细胞骨架网络的结构。 以及解释这些电机中有多少沿着其细丝轨道移动的分子细节 在细胞环境中，大多数这些马达与复杂的调节机制协同作用。 将它们与各自的货物连接起来，调节它们的运动特性，并指示 目前尚不清楚该机器的个体运动输出如何。 此外，许多货物是在细胞的复杂环境中清晰且难以解剖的。 由相反极性的电机同时移动，这一过程称为双向传输。 各个电机被招募到货物、激活并与其他类别的电机集成 此外，不同电机的活动被利用来对该领域提出了巨大的挑战。 构建和维持关键的细胞骨架结构，例如有丝分裂纺锤体、纤毛和分裂 犁沟如何协调运动和调节活动来驱动此类自组装。 目前，结构是正常理解和患病细胞生理学的重大障碍。 该应用程序旨在开发新的测定方法和工具来研究运动募集的复杂性 和调节、货物的双向运输以及细胞骨架结构的自组装 由电机和相关分子驱动的我们将生物化学和单分子相结合的方法。 体外重构的分子分析，测试分子功能，并转化我们的 测试这些重组产生的假设的体内系统的发现将开放 我们建议： 1）重新构建和研究招聘， 与膜货物结合的细胞质动力蛋白和驱动蛋白马达的调节和运动 通过已知将这些马达与内吞细胞连接起来的内源性 Rab GTPase 机制 细胞中的囊泡和线粒体，2) 重建和研究动力蛋白和驱动蛋白的功能 这些广泛的目标建立并扩展了我们的目标。 剖析细胞质动力蛋白马达调节机制的专业知识和之前的工作， 旨在提供有助于剖析复杂运动功能的强大新工具。 将阐明驱动细胞稳态的基本分子和细胞生物学原理 提供对分子运动故障引起的病理机制的深入了解。