Structure and functions of the actin cytoskeleton

肌动蛋白细胞骨架的结构和功能

• 批准号: 10470372
• 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/10470372
• 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; base; 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.

项目概要/摘要 肌动蛋白细胞骨架是一种主要的细胞成分，几乎在细胞的各个方面都具有关键功能 生理学，包括细胞运动、形状和力学、细胞和组织形态发生、细胞间和细胞基质 肌动蛋白细胞骨架结构、功能的相互作用和动力学。 和/或动力学对人类病理学有重大贡献，特别是对癌症和神经退行性疾病， 肌动蛋白细胞骨架由于其能力而在细胞中发挥着不可或缺的作用。 以多种不同的组合产生大的推力、拉力和阻力来执行这些操作。 肌动蛋白丝具有多种功能，由多种辅助蛋白组织成多种结构阵列。 尽管进行了广泛的研究，但这些基于肌动蛋白的分子机器的确切组织经常是不明确的。 实现这一目标的主要障碍是解决肌动蛋白细胞骨架结构的困难。 如果不了解结构，就无法了解机械的功能。 我的实验室使用了一种独特的方法来克服这个问题。 复制电子显微镜（PREM），它独特地能够结合高分辨率成像 全覆盖整个细胞的细胞骨架，并有效地将细胞骨架结构与活体相关联 在 PREM 的帮助下，我的实验室在理解细胞动力学方面做出了多项基础贡献。 在此应用中，我们提出了一系列通用和特化细胞类型中的细胞骨架功能。 未来五年的进程，以解决代表四个主要类别的以下问题 肌动蛋白细胞骨架的功能：（1）突出——微管如何调节肌动蛋白的突出活性 定向迁移的细胞骨架；(2) 收缩——非肌肉肌球蛋白 II 之间如何相互作用 (3) 细胞力学——差异如何 肌动蛋白皮层的分子结构与正常和不同的机械特性有关 癌细胞；(4) 膜动力学——分支肌动蛋白网络如何促进网格蛋白包被的内陷 除了广泛的其他细胞生物学、成像、 功能、生物化学和分子生物学方法使我们处于独特的地位，可以显着推进 反过来，我们对肌动蛋白细胞骨架功能的理解可能会提供重要的新见解。 研究如何对抗与肌动蛋白细胞骨架功能障碍相关的人类疾病。