The Molecular Basis for Myosin Regulation

肌球蛋白调节的分子基础

基本信息

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
项目类别：
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

项目摘要

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一个关键功能是在需要时移动的能力。这些运动——细胞内运输、细胞分裂、肌肉收缩和细胞运动——由分子机器驱动，分子机器发挥作用考虑到它们的直径只有几纳米，其力量是惊人的。鉴于电机的多样性细胞中的蛋白质，一个关键问题是电机在移动货物和应用时如何合作和竞争力量。一种新兴的范例是专用电机的概念，或者经过微调以执行特定操作的电机。具体功能。尽管这些运动蛋白很重要，但人们对它们的个体知之甚少适应性以及这些适应性与细胞中发现的运动模式有何关系。这项工作重点关注肌球蛋白 6 和 myosin-10 及其独特的细胞调节形式。 Myosin-6 在细胞器中发挥重要作用形态、细胞形态、胞质分裂、自噬和内吞作用，而肌球蛋白 10 则提供必需的整合素、钙粘蛋白和 netrin 受体等货物到达细胞前缘的丝状伪足。两者都是由于其在膜运输、细胞迁移和转移中的作用，在肿瘤中过度表达。这这项工作将开发新方法来控制肌球蛋白 6 并分离其在细胞中的活性。主要之一方法是用光隔离肌球蛋白 6，使细胞具有可光学切换的斯涅耳氏囊 waltzer（肌球蛋白 6 无效）表型。实验旨在确定肌球蛋白 6 何时充当乘客，锚或单元中的运输装置，如果需要从一个货物适配器直接切换到另一个功能。拟议的工作还将研究货物结合和肌球蛋白四级结构如何调整肌球蛋白10的运动性。开发了一种综合方法，将结构研究与功能研究相结合重构和单分子运动测定。该提案将测试以下假设：货物和细胞外配体结合是肌球蛋白 10 激活所必需的，并且该货物可以引导肌球蛋白 10 将一种类型的肌动蛋白网络连接到另一种类型的肌动蛋白网络。完成本研究将全面了解如何细胞骨架运动蛋白被激活和调节以执行细胞中的不同任务。运动蛋白调节是这一过程具有重要的生物学基础，但人们对其知之甚少。这项工作将指导未来的努力了解多种情况下的激活。