Cytoskeletal Motors and Scaffolds in Membrane Dynamics and Motility

描述(申请人提供)：微管和肌动蛋白细胞骨架对细胞内膜的移动、极化、分选和形态形成是必不可少的。分子马达、支架蛋白和相关的细丝被招募到膜上，为不同的运输活动提供动力，这些活动具有不同的力、功率、动力学和运输要求。确定贩运的机制需要了解不同的运动异构体和家族如何单独和在团队中工作，以及马达和细丝如何与将它们连接到膜的支架蛋白和适配器一起工作。确定这些机制需要详细了解：细胞骨架和膜的细胞组织和动力学；不同马达和细丝的空间、动力学和力学关系；细胞骨架-膜组件的结构和生物物理性质；以及定义马达和支架在工作条件下运行的能力和机制的生物物理参数。为此，我们组建了一个非常强大的科学团队，拥有细胞生物学、生物化学、结构生物学、结构动力学和技术开发方面的专业知识，以确定细胞骨架和分子马达在贩运中的作用。我们的团队包括使用最先进的成像、单分子和结构技术来发现细胞骨架蛋白如何在复杂的细胞事件中发挥作用的先驱。高分辨率活细胞显微镜、利用微细加工和介电技术重建的细胞骨架几何结构、X射线结晶学、纳米分辨荧光跟踪、单分子荧光偏振、光学捕获和先进的生化技术将在高度协作的研究中应用，以了解发动机、支架和细丝如何共同为膜动力学提供动力。项目和调查人员是相互依存的，通过研究目标和共同技术紧密联系在一起，制定这些目标是为了利用小组成员的独特优势，同时利用小组之间的广泛协同作用。我们将集中于以下四个目标：(1)研究细胞器运输和膜重塑中的分子马达动力学；(2)研究细胞器形态发生和运动中细胞骨架-膜支架的结构、生化和细胞性质；(3)发现膜相关马达和支架的机械和生化适应；(4)研究肌球蛋白、动力蛋白和运动集合的结构动力学。

DESCRIPTION (provided by applicant): The microtubule and actin cytoskeletons are essential for the movement, polarization, sorting, and morphogenesis of intracellular membrane compartments. Molecular motors, scaffolding proteins, and associated filaments are recruited to membranes to power diverse trafficking events that have different force, power, kinetic, and transport requirements. Defining the mechanisms of trafficking requires understanding how different motor isoforms and families work alone and in teams, and how motors and filaments work with the scaffolding proteins and adapters that link them to membranes. Determining these mechanisms requires a detailed understanding of: the cellular organization and dynamics of the cytoskeleton and membranes; the spatial, kinetic, and mechanical relationship of different motors and filaments; the structural and biophysical properties of cytoskeletal-membrane assemblies; and the biophysical parameters that define the capabilities and mechanisms of motors and scaffolds when operating under working conditions. To this end, we assembled an extraordinarily strong scientific team with expertise in cell biology, biochemistry, structural biology, structural dynamics, and technology-development to define the role of the cytoskeleton and molecular motors in trafficking. Our team includes pioneers in the use of state-of-the-art imaging, single-molecule, and structural techniques to discover how cytoskeletal proteins function in complex cellular events. High-resolution live-cell microscopy, reconstituted cytoskeletal geometries using microfabrication and dielectrophoresis, X-ray crystallography, nanometer-resolved fluorescence tracking, single-molecule fluorescence polarization, optical trapping, and advanced biochemical techniques will be applied in highly collaborative studies to understand how motors, scaffolds, and filaments work together to power membrane dynamics. The projects and investigators are interdependent and are closely linked through research goals and common technologies, and the Aims were formulated to capitalize on the unique strengths of the team members while taking advantage of extensive synergies between the groups. We will focus on the following four Aims: (1) Investigate the Dynamics of Molecular Motors in Organelle Transport and Membrane Remodeling; (2) Investigate the Structural, Biochemical, and Cellular Properties of Cytoskeleton-Membrane Scaffolds in Organelle Morphogenesis and Motility; (3) Discover the Mechanical and Biochemical Adaptations of Membrane- Associated Motors and Scaffolds; (4) Investigate the Structural Dynamics of Myosin, Dynein and Motor Collections.