Converting cytoskeletal forces into biochemical signals

将细胞骨架力转化为生化信号

基本信息

批准号：
10655891
负责人：
GREGORY M ALUSHIN
金额：
$ 33.9万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-15 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10655891
关键词：
Actins Binding Biochemical Biological Process Biophysical Process Biophysics Bundling Cell Cycle Arrest Cell Death Cell Line Cell Nucleus Cell physiology Cell-Matrix Junction Cells Cellular Assay Clustered Regularly Interspaced Short Palindromic Repeats Complex Coupled Cryo-electron tomography Cryoelectron Microscopy Cytoplasm Cytoskeletal Modeling Cytoskeleton Data Development Disease Dissection Environment Evaluation Event FHL1 gene Filament Fluorescence Microscopy Functional disorder Gene Expression Gene Expression Regulation Generations Genes Homeostasis Individual Knock-out Link Lobular Neoplasia Malignant Neoplasms Mechanics Mediating Membrane Methods Microfilaments Molecular Structure Motor Muscular Dystrophies Myosin ATPase Nuclear Outcome Output Pathway interactions Physical condensation Polymers Protein Engineering Proteins Reporting Resolution Role Rupture Signal Transduction Site Stress Fibers Structure Technology Testing Therapeutic Tissues Transcription Factor AP-1 Visualization Work ZYX gene alpha Actinin force feedback immune function in vivo inhibitor innovation mechanical signal mechanotransduction nanometer resolution polymerization prevent protein crosslink reconstitution reconstruction recruit repaired therapeutic target transcriptome sequencing transmission process vasodilator-stimulated phosphoprotein

项目摘要

PROJECT SUMMARY Cells perceive mechanical cues in their local environments, which must be converted into intracellular biochemical signals to modulate cellular physiology and control gene expression. There is increasing appreciation for mechanical signal transduction’s (“mechanotransduction”) critical role in development and its dysfunction in disease states such as cancer. However, in contrast to canonical signal transduction, cellular force sensing is poorly understood, hampering efforts to define mechanistically distinct mechanotransduction pathways, delineate their specific biological functions, and target them therapeutically. The actin cytoskeleton, a network of dynamic actin filaments, myosin motor proteins, and hundreds of associated factors, enables cells to mechanically interface with their surroundings. The cytoskeleton is classically understood to serve as a force generation and transmission apparatus that indirectly facilitates mechano- transduction through its physical linkages to membrane-anchored sites which mediate force signal conversion (e.g. cell-cell and cell-matrix adhesions). However, we and others have recently reported direct binding of soluble cytosolic proteins containing tandem arrays of LIM (LIN-11, Isl-1 & Mec-3) domains to tensed actin filaments, suggesting that the cytoskeleton itself may have the capacity to transduce forces into biochemical signals. Here I propose to test the hypothesis that force-activated actin binding by distinct LIM proteins is upstream of functionally discrete downstream mechanotransduction pathways. Through cellular assays and biophysical reconstitution, we will investigate how the representative force-activated actin binding LIM proteins zyxin (Aim 1) and FHL1/2 (Four-and-a-Half LIM domains 1/2, Aim 2) mediate distinct downstream functions in cytoplasmic cytoskeletal damage repair and nuclear gene expression regulation, respectively. We will then innovatively interface these approaches with cryo-electron microscopy (cryo-EM) to visualize force-activated actin binding by LIM proteins in structural detail (Aim 3). Our studies will establish how a conserved mechanism of force transduction through LIM domains is linked to distinct downstream signaling outcomes, which is likely to reveal general principles underlying the modular organization of cytoskeletal mechanical signaling networks. In the longer term, this work will enable precision dissection of context-specific biological functions of LIM proteins in vivo, facilitating rigorous evaluation of their potential as therapeutic targets.

项目概要细胞感知局部环境中的机械信号，必须将其转化为细胞内信号调节细胞生理学和控制基因表达的生化信号越来越多。认识机械信号转导（“机械转导”）在发育中的关键作用及其然而，与典型的信号转导相反，细胞力。人们对传感知之甚少，阻碍了定义机械上不同的机械转导的努力途径，描述其特定的生物学功能，并针对它们进行治疗。肌动蛋白细胞骨架，动态肌动蛋白丝、肌球蛋白运动蛋白和数百个的网络相关因素使细胞能够与周围环境进行机械连接。被理解为作为力的产生和传输装置，间接促进机械- 通过与介导力信号转换的膜锚定位点的物理连接进行转导（例如细胞-细胞和细胞-基质粘附）然而，我们和其他人最近报道了可溶性的直接结合。含有 LIM（LIN-11、Isl-1 和 Mec-3）结构域与张紧肌动蛋白丝串联阵列的胞质蛋白，这表明细胞骨架本身可能具有将力转化为生化信号的能力。我建议检验以下假设：不同 LIM 蛋白的强制激活肌动蛋白结合位于通过细胞测定和生物物理功能离散的下游机械转导途径。重建后，我们将研究代表性的力激活肌动蛋白如何结合 LIM 蛋白 zyxin（目标 1）和 FHL1/2（四个半 LIM 结构域 1/2，目标 2）介导细胞质中不同的下游功能然后我们将分别创新性地进行细胞骨架损伤修复和核基因表达调控。将这些方法与冷冻电子显微镜（cryo-EM）结合起来，通过以下方式可视化力激活的肌动蛋白结合 LIM 蛋白的结构细节（目标 3）。通过 LIM 结构域的转导与不同的下游信号传导结果相关，这可能揭示细胞骨架机械信号网络模块化组织的一般原则。从长远来看，这项工作将能够精确解析 LIM 蛋白的特定生物功能体内，促进对其作为治疗靶点的潜力进行严格评估。