Converting cytoskeletal forces into biochemical signals

将细胞骨架力转化为生化信号

• 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
• 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

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.

项目摘要 细胞在其局部环境中感知机械提示，这些机械提示必须被转化为细胞内信号。 生物化学信号调节细胞生理学和控制基因表达。人们日益 对机械信号转导（“机械转导”）在发育中的关键作用及其 在疾病状态如癌症中的功能障碍。然而，与典型的信号转导相反，细胞力 对感觉的理解很少，阻碍了对机械性不同的机械转导的定义 我们可以通过这些途径，描述其特定的生物学功能，并在治疗上靶向它们。 肌动蛋白细胞骨架，动态肌动蛋白丝网络，肌球蛋白马达蛋白，以及数百种 相关因子，使细胞能够与周围环境机械地相互作用。细胞骨架是典型的 被理解为用作力产生和传递装置，其间接地促进机械运动。 通过其与介导力信号转换的膜锚定位点的物理连接进行转导 (e.g.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 含有LIM（LIN-11，Isl-1和Mec-3）结构域串联阵列的细胞溶质蛋白质， 这表明细胞骨架本身可能有能力将力转换为生化信号。这里 我建议测试的假设，力激活肌动蛋白结合不同的LIM蛋白的上游， 功能离散的下游机械转导途径。通过细胞分析和生物物理 重建，我们将研究如何代表力激活肌动蛋白结合LIM蛋白zyxin（目的1） 和FHL 1/2（Four-and-a-Half LIM domain 1/2，Aim 2）介导细胞质中不同的下游功能， 细胞骨架损伤修复和核基因表达调控。我们将创新地 将这些方法与冷冻电子显微镜（cryo-EM）相结合， LIM蛋白的结构细节（目的3）。我们的研究将证明力的守恒机制 通过LIM结构域的转导与不同的下游信号转导结果有关，这可能揭示了 细胞骨架机械信号网络模块化组织的基本原理。在 从长远来看，这项工作将能够精确解剖LIM蛋白的特定生物学功能， 体内，促进其作为治疗靶点的潜力的严格评估。