Regulation and Function of Intermediate Filaments in Cell Mechanics

细胞力学中中间丝的调节和功能

• 批准号: 8665988
• 负责人: ROBERT D GOLDMAN
• 金额: $ 174.85万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8665988
• 关键词: Affect; Binding; Biological Assay; Bundling; Cell Shape; Cell surface; Cells; Chemicals; Complex; Connecticut; Cytoplasm; Cytoskeleton; Disease Attributes; Elements; Endothelial Cells; Event; Family; Fibroblasts; Genes; Growth Factor; Guanosine Triphosphate Phosphohydrolases; In Vitro; Intermediate Filament Proteins; Intermediate Filaments; Life; Link; Mammalian Cell; Mechanical Stress; Mechanics; Membrane; Microfilaments; Microtubules; Molecular Motors; Monitor; Motor; Movement; Mutation; Pennsylvania; Peptides; Phosphorylation; Phosphorylation Site; Play; Principal Investigator; Process; Program Research Project Grants; Property; Proteins; Regulation; Research; Role; Series; Serum; Signal Transduction; Signal Transduction Pathway; Site; Structural Protein; Structure; System; Testing; Universities; Vimentin; X-Ray Crystallography; biophysical properties; biophysical techniques; cell motility; design; dimer; insight; mimetics; particle; research study; response; rho; shear stress

DESCRIPTION (Provided by applicant): In this Program Project Grant our overarching hypothesis is that alterations in the assembly states and mechanical properties of cytoskeletal IF, specifically the type III IF composed of vimentin (VIF), play important roles in regulating the micromechanical properties of cells in response to mechano- and chemo- signaling. These studies are critically important as IF are major elements of the cytoskeletal system of mammalian cells and yet their specific functions in cell motility remain unknown. The 6 Project Leaders, and their research aims are as follows: R. Goldman, Northwestern University, will determine how the vimentin IF (VIF) system changes in response to mechanical and chemical signals, and will purify and characterize the vimentin precursors that form as VIF assemble and disassemble. V. Gelfand, Northwestern University will determine the relationship between VIF with microtubules and actin filaments, identify the molecular motors responsible for translocating VIF precursors, and investigate how VIF affect cytoskeletal dynamics and microtubule (MT) dynamics. G. Danuser, Harvard University, will test the hypotheses that VIF network formation involves a spatially distributed assembly line, that this process involves molecular motors, and that VIF network assembly modulates assembly of MT and microfilaments (MF). D. Weitz, Harvard University, will determine the micromechanical properties of in vitro assembled VIF networks prepared from purified vimentin, native VIF networks isolated from cells and in living cells. P. Janmey, University of Pennsylvania, will determine how the VIF network contributes to the micromechanical properties of living cells and how this changes in response to substrate stiffness and external forces. P. Burkhard, University of Connecticut, will use biophysical techniques and X-ray crystallography to determine the structure and biophysical properties of the vimentin dimer and study the formation of VIF assembly intermediates. In all of these studies, the regulatory function of different posttranslational phosphorylation events will be examined.

描述（由申请人提供）：在此计划项目中，我们的总体假设是，if if的组装状态和机械特性的改变，如果是由波形蛋白（VIF）组成的III型，则在调节机械和化学信号响应细胞的微机械特性中起着重要作用。这些研究至关重要，好像是哺乳动物细胞的细胞骨架系统的主要元素，而它们在细胞运动中的特定功能仍然未知。 6个项目负责人及其研究的目的如下：西北大学的R. Goldman将确定if（VIF）系统对机械和化学信号的响应发生变化，并将纯化和表征形成VIF组装和拆卸的波形蛋白前体的变化。 V. Gelfand，西北大学将确定VIF与微管和肌动蛋白丝之间的关系，确定负责易位VIF前体的分子电机，并研究VIF如何影响细胞骨架动力学和微管（MT）动力学。哈佛大学的G. Danuser将测试VIF网络形成的假设涉及空间分布的组装线，该过程涉及分子电机，并且VIF网络组装调节MT和微丝（MF）的组装。哈佛大学的D. Weitz将确定由纯化的波形蛋白，从细胞和活细胞中分离的天然VIF网络制备的体外组装的VIF网络的微力特性。 P. Janmey，宾夕法尼亚大学，将确定VIF网络如何促进活细胞的微力特性，以及这如何响应底物刚度和外部力。康涅狄格大学的P. Burkhard将使用生物物理技术和X射线晶体学来确定波形蛋白二聚体的结构和生物物理特性，并研究VIF组装中间体的形成。在所有这些研究中，将检查不同翻译后磷酸化事件的调节功能。