Structure and functions of the actin cytoskeleton

肌动蛋白细胞骨架的结构和功能

• 批准号: 10667325
• 负责人: Tatyana Svitkina
• 金额: $ 40.63万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-16 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667325
• 关键词: Actins; Address; Architecture; Biochemical; Biological; Cardiovascular Diseases; Categories; Cell physiology; Cells; Clathrin; Cytoskeleton; Electron Microscopy; Filament; Goals; Human Pathology; Image; Immune System Diseases; Knowledge; Link; Malignant Neoplasms; Mechanics; Membrane; Methods; Microfilaments; Microtubules; Molecular; Morphogenesis; Myosin Type II; Neurodegenerative Disorders; Normal Cell; Organelles; Platinum; Play; Positioning Attribute; Proteins; Research; Resistance; Role; Shapes; Structure; System; Tissues; cancer cell; cell motility; cell type; combat; diagnostic strategy; high resolution imaging; human disease; insight; mechanical properties; migration; non-muscle myosin; paralogous gene; treatment strategy; virtual

PROJECT SUMMARY/ABSTRACT The actin cytoskeleton is a major cellular component with key functions in virtually every aspect of cell physiology including cell motility, shape and mechanics, cell and tissue morphogenesis, cell-cell and cell-matrix interactions, and dynamics of membrane organelle. Aberrations in actin cytoskeleton structure, functions and/or dynamics contribute significantly to human pathologies, especially to cancer and neurodegenerative, immune and cardiovascular diseases. The actin cytoskeleton plays indispensable roles in cells due to its ability to generate large pushing, pulling and resistance forces in many different combinations. To perform these diverse functions, actin filaments are organized into diverse structural arrays by multiple accessory proteins. Despite extensive research, the exact organization of these actin-based molecular machineries is frequently unknown. The main barrier toward this goal is the difficulty of resolving actin cytoskeleton architecture at a single-filament level. Without knowledge of the structure, functional understanding of the machinery is incomplete. My lab uses a distinctive approach to overcome this problem. We take advantage of platinum replica electron microscopy (PREM), which is uniquely able to combine high resolution imaging of the cytoskeleton with full coverage of the whole cell and to efficiently correlate the cytoskeleton structure with live cell dynamics. With help of PREM, my lab has made multiple fundamental contributions toward understanding of cytoskeleton functions in a range of generic and specialized cell types. In this application, we propose in the course of the next five years to address the following questions representing each of the four major categories of actin cytoskeleton functions: (1) Protrusion – How microtubules regulate protrusive activity of the actin cytoskeleton for directional migration; (2) Contraction – How an interplay between nonmuscle myosin II paralogs regulates polarized subcellular distribution of contractile forces; (3) Cell mechanics – How differences in the molecular architecture of the actin cortex are linked to different mechanical properties of normal and cancer cells; (4) Membrane dynamics – How branched actin networks promote invagination of clathrin-coated membrane domains. Our expertise in PREM in addition to a broad range of other cell biological, imaging, functional, biochemical, and molecular biological methods puts us in unique position to significantly advance our understanding of actin cytoskeleton functions. In turn, this knowledge may provide important new insights into how to combat human diseases associated with actin cytoskeleton malfunctions.

项目摘要/摘要 肌动蛋白细胞骨架是一个主要的细胞分量 生理学，包括细胞运动，形状和力学，细胞和组织形态发生，细胞 - 细胞和细胞矩阵 相互作用和膜细胞器的动力学。肌动蛋白细胞骨架结构的畸变，功能 和/或动力学对人类病理有显着贡献，尤其是对癌症和神经退行性的病理学 免疫和心血管疾病。肌动蛋白细胞骨架由于能力而在细胞中起着不可或缺的作用 以许多不同的组合产生大型推动，拉力和阻力。执行这些 多种功能，肌动蛋白丝由多种辅助蛋白组织成潜水的结构阵列。 尽管进行了广泛的研究，但这些基于肌动蛋白的分子机的确切组织经常是 未知。实现这一目标的主要障碍是难以解决肌动蛋白细胞骨架建筑 单丝水平。在不了解结构的情况下，对机械的功能理解是 不完整。我的实验室使用一种独特的方法来克服这个问题。我们利用白金 复制电子显微镜（PER），它具有独特的能力，能够结合高分辨率成像 细胞骨架具有全部覆盖整个细胞的覆盖范围，并有效地将细胞骨架结构与活体 细胞动力学。在Prem的帮助下，我的实验室为理解做出了多种基本贡献 细胞骨架在一系列通用和专业细胞类型中的功能。在此应用程序中，我们在 未来五年的课程解决以下问题，代表四个主要类别中的每个类别 肌动蛋白细胞骨架功能：（1）突出 - 微管如何调节肌动蛋白的爆发活性 用于定向迁移的细胞骨架； （2）收缩 - 非肌肉肌球蛋白II之间的相互作用如何 旁系同源物调节收缩力的极化亚细胞分布； （3）细胞力学 - 差异如何 在肌动蛋白皮质的分子结构中，与正常和 癌细胞； （4）膜动力学 - 分支肌动蛋白网络如何促进网状蛋白涂层的内幕 膜域。除了广泛的其他细胞生物学成像外，我们在PER上的专业知识， 功能，生化和分子生物学方法使我们处于独特的位置 我们对肌动蛋白细胞骨架功能的理解。反过来，这些知识可能会提供重要的新见解 如何打击与肌动蛋白细胞骨架故障相关的人类疾病。