The Molecular Basis for Myosin Regulation

肌球蛋白调节的分子基础

• 批准号: 9767242
• 负责人: Ronald S Rock
• 金额: $ 34.36万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767242
• 关键词: Actins; Address; Affect; Amaze; Autophagocytosis; Behavior; Binding; Biological; Biological Assay; Biological Models; Cadherins; Cell Shape; Cell Surface Receptors; Cell division; Cell physiology; Cells; Cellular Morphology; Complex; Cytokinesis; Cytoskeletal Modeling; Data; Defect; Detection; Endocytic Vesicle; Endocytosis; Ensure; Filament; Filopodia; Future; Goals; Golgi Apparatus; Hand; In Vitro; Individual; Integrins; Kinetics; Knock-out; Knowledge; Length; Life; Ligand Binding; Ligands; Light; Malignant Neoplasms; Mechanics; Membrane; Methods; Mitochondria; Molecular; Molecular Machines; Morphology; Motor; Motor Activity; Movement; Muscle Contraction; Myosin ATPase; Neoplasm Metastasis; Optics; Organelles; Organism; Pattern; Phenotype; Play; Process; Proteins; Regulation; Role; Signal Transduction; Site; Structure; System; Techniques; Testing; Vesicle; Work; cancer cell; cell growth regulation; cell motility; daughter cell; design; dimer; experimental study; extracellular; fascin; nanometer; netrin receptor; novel; novel strategies; optical switch; optogenetics; overexpression; reconstitution; recruit; single molecule; tool; trafficking; tumor

Abstract A critical function for all living organisms is the ability to move when needed. These movements--intracellular trafficking, cell division, muscle contraction, and cell motility-- are driven by molecular machines that exert an amazing amount of force considering that they are only a few nanometers across. Given the variety of motor proteins in the cell, a key question is how motors cooperate and compete while moving cargoes and applying forces. An emerging paradigm is the notion of specialized motors, or motors that are fine-tuned to perform a specific function. Despite the importance of these motor proteins, relatively little is known about their individual adaptations and how these relate to the motility patterns found in the cell. This work focuses on myosin-6 and myosin-10 and their unique forms of cellular regulation. Myosin-6 plays essential roles in organelle morphology, cell morphology, cytokinesis, autophagy, and endocytosis, while myosin-10 delivers essential cargoes such as integrins, cadherins and netrin receptors to filopodia at the leading edge of the cell. Both are overexpressed in tumors, owing to their roles in membrane trafficking, cell migration, and metastasis. The work will develop new approaches to control myosin-6 and isolate its activity in cells. One of the main approaches will be to sequester myosin-6 with light, making cells that have an optically switchable Snell's waltzer (myosin-6 null) phenotype. Experiments are designed to identify when myosin-6 acts as a passenger, an anchor, or a transporter in the cells, and if direct handoff from one cargo adaptor to another is required for function. The proposed work will also investigate how cargo binding and myosin quaternary structure tune myosin-10's motility. An integrated approach is developed, combining structural studies with functional reconstitution and single molecule motility assays. This proposal will test the hypotheses that cargo and extracellular ligand binding are both required for myosin-10 activation, and that cargo can steer myosin-10 from one type of actin network to another. Completion of this study will yield a comprehensive view of how cytoskeletal motor proteins are activated and regulated for distinct tasks in the cell. Motor protein regulation is a process of fundamental biological importance, but is poorly understood. This work will direct future efforts to understand activation in multiple contexts.

抽象的 所有生物体的关键功能是在需要时移动的能力。这些运动 - 细胞 运输，细胞分裂，肌肉收缩和细胞运动性 - 由分子机驱动 考虑到它们只是少数纳米的惊人力量。鉴于电动机的种类 细胞中的蛋白质，一个关键的问题是电动机在移动货物和申请时如何合作和竞争 力量。新兴范式是专业电动机的概念，或者是对执行的电动机的概念 特定功能。尽管这些运动蛋白很重要，但对他们的个体知之甚少 适应以及这些与细胞中的运动模式如何相关。这项工作的重点是肌球蛋白6和 肌球蛋白10及其独特的细胞调节形式。肌球蛋白6在细胞器中扮演重要角色 形态，细胞形态，细胞因子，自噬和内吞作用，而肌球蛋白10可提供必需品 整合素，钙粘蛋白和Netrin受体等货物在细胞前缘的丝状受体。两者都是 由于它们在膜运输，细胞迁移和转移中的作用，在肿瘤中过表达。这 工作将开发新的方法来控制肌球蛋白6并隔离其在细胞中的活性。主要之一 方法将是用光隔离肌球蛋白6，使单元具有光学上可切换的Snell的细胞 Waltzer（肌球蛋白-6 NULL）表型。实验旨在识别肌球蛋白6何时充当乘客， 单元格中的锚或转运蛋白，如果需要从一个货物适配器直接交接到另一个货物适配器 功能。拟议的工作还将调查货物结合和肌球蛋白第四纪结构如何调整 肌球蛋白10的运动。开发了一种综合方法，将结构研究与功能相结合 重构和单分子运动测定法。该提案将检验货物和 细胞外配体结合都是肌球蛋白10激活所必需的，并且货物可以从 一种类型的肌动蛋白网络到另一种类型。这项研究的完成将对如何进行全面的看法 细胞骨架运动蛋白被激活并调节细胞中的不同任务。运动蛋白调节是 基本生物学重要性的过程，但知之甚少。这项工作将把未来的努力引导到 了解多种情况下的激活。