Coordination of molecular motor activity in intracellular transport and assembly of cytoskeletal archit

细胞内转运和细胞骨架组装中分子马达活性的协调 建筑。 私家侦探-理查德·J·麦肯尼 研究总结 在真核生物中，细胞内转运是细胞内平衡所必需的。这一过程被执行 通过分子马达将ATP水解产生的化学能转化为沿肌动蛋白的运动 和微管细胞骨架网络。几十年的研究揭示了结构和分子 详细解释了有多少这样的马达沿着它们的丝状轨迹运动。在蜂窝环境中， 这些发动机中的大多数都与复杂的监管机制协同工作，这些监管机制将它们与其 各自的货物，调节它们的运动属性，并决定时空活动。多么 单个电机的输出是由这台机器控制的，目前还不清楚，很难剖析 细胞的复杂环境。此外，许多货物的移动是由于……的协同行动 在一种称为双向传输的过程中，同时绑定相反极性的电机。多么 单独的马达被招募到货物上，被激活，并与其他类别的马达集成 对该领域提出了一个巨大的公开挑战。重要的是，这一过程中的缺陷导致了各种各样的 因此，这些问题直接关系到人类健康。 微管网络组织、动力学和沿微管的运动活动都是 受到非酶微管相关蛋白(MAP)的冲击，MAP动态结合到 微管。MAP的特定功能、分子特性和动力学仍然存在 开发不足。Tau家族的地图对人类健康至关重要，因为tau很好- 以在许多人类神经退行性疾病中形成不可溶的包涵体/聚集体为特征 例如阿尔茨海默氏症和皮克氏病。尽管他们的身份在40多年前就被确认了， Tau家族图谱的具体分子功能和分子性质尚不清楚。这 应用寻求开发新的分析方法和工具来研究微管电机的复杂性 监管、货物双向运输以及由马达和地图驱动的细胞骨架功能。 我们将生物化学和单分子分析相结合的体外重组方法 这测试了分子的功能，并将我们的发现转化为体内系统来测试假说 由这些重组产生的，将开辟卓有成效的长期研究途径。我们建议 目的：1)重建和研究胞浆动力蛋白的募集、调节和运动 与天然细胞支架分子结合的动蛋白马达，2)重建和研究 Tau家族图谱的进化功能、动力学和病理行为。这些宏大的目标 以我们的专业知识和以前的工作为基础，剖析 细胞质动力蛋白马达，旨在提供对解剖有用的强大的新工具 复杂的运动功能。我们的工作将阐明基本的分子和细胞生物学原理 促进细胞动态平衡，并提供对以下病理机制的见解 分子马达故障。

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. 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 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 by the concerted action of simultaneously bound, opposite polarity motors, in a process called bidirectional transport. How individual motors are recruited to cargo, activated, and integrated with other classes of motors presents a large open challenge to the field. Importantly, defects in this process lead to a wide variety of human diseases and these questions are thus directly related to human health. Microtubule network organization, dynamics, and motor activity along microtubules are all impinged upon by non-enzymatic microtubule-associated proteins (MAPs) that dynamically bind to microtubules. The specific functions, molecular properties, and dynamics of MAPs remains underexplored. The tau family of MAPs are critically important for human health, as tau is well- characterized to form insoluble inclusions/aggregates in a host of human neurodegenerative diseases such as Alzheimer’s disease and Pick’s disease. Despite their identification over four decades ago, the specific molecular functions and molecular properties of tau family MAPs remains unclear. This application seeks to develop novel assays and tools to study the complexity of microtubule motor regulation, bidirectional transport of cargos, and cytoskeletal functions driven by motors and MAPs. 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 native cellular cargo scaffolding molecules, and 2) Reconstitute and study evolutionary functions, dynamics, and pathological behaviors of tau family MAPs. These broad goals build and expand upon 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.