Integrin-filamin Interactions in Migration and Signaling

整合素-细丝蛋白在迁移和信号转导中的相互作用

• 批准号: 6928000
• 负责人: DAVID A CALDERWOOD
• 金额: $ 28.61万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6928000
• 关键词: CHO cells; SDS polyacrylamide gel electrophoresis; X ray crystallography; actin binding protein; affinity chromatography; binding sites; biological signal transduction; cell adhesion molecules; cell migration; circular dichroism; green fluorescent proteins; immunoprecipitation; integrins; nuclear magnetic resonance spectroscopy; phosphorylation; polymerase chain reaction; protein binding; protein protein interaction; proteolysis; surface plasmon resonance

DESCRIPTION (provided by applicant): Support is requested to analyze the interaction between the actin crosslinking protein filamin and integrin beta subunit cytoplasmic tails; the effect this interaction has on integrin signaling; the mechanisms by which integrin-filamin interactions inhibit cell migration; and how the interaction is regulated in cells. Heterodimeric integrin adhesion receptors mediate cell-extracellular matrix and cell-cell adhesion events throughout development, during hemostasis and in the response to injury and infection. The binding of specific signaling and cytoskeletal proteins to integrin beta tails controls cell adhesion, bidirectional integrin signaling and regulates cell motility. Integrin-mediated cell migration is essential for development, the immune response, and for tissue remodeling and wound healing. Integrin-mediated cell migration also contributes to a number of disease states and is required for tumor metastasis, inflammation and the formation of atherosclerotic plaques. The increased association of filamin with integrin beta tails inhibits cell migration. The applicant hypothesizes that regulated association of filamin with integrin beta tails mediates distinct integrin functions and acts as a brake on cell migration. To test this hypothesis he aims to map the integrin binding site in filamin and identify mutations that selectively inhibit integrin binding. He will use these mutants and previously identified integrin mutants that up- or down-regulate filamin binding to characterize the effect of integrin-filamin interactions on integrin-mediated signal transduction. He will then investigate whether the inhibition of cell migration following increased filamin binding to integrins is due to the observed effects on integrin signaling, to local filamin-mediated effects on actin filament crosslinking or to a combination of both processes. Finally he will assess effect of filamin proteolysis, filamin or integrin phosphorylation and competition between integrins and other filamin-binding proteins on the association of filamin with integrin beta tails. These studies will characterize the regulation and mechanism of action of a pathway that controls cell migration. Cell migration is a tightly controlled process that plays a central role in many biological phenomena. Therefore these studies will provide insight into a process essential during health and disease and may identify novel therapeutic targets for treatment of cancers, arthritis and atherosclerosis.

描述（由申请人提供）：要求支持分析肌动蛋白交联蛋白丝蛋白和整合素β亚基细胞质尾部之间的相互作用；这种相互作用对整合素信号传导的影响；整合素-丝蛋白相互作用抑制细胞迁移的机制；以及这种相互作用是如何在细胞中被调节的。异二聚体整合素粘附受体介导细胞-细胞外基质和细胞-细胞粘附事件在整个发育过程中，在止血和对损伤和感染的反应中。特异性信号和细胞骨架蛋白与整合素尾部的结合控制细胞粘附、双向整合素信号和调节细胞运动。整合素介导的细胞迁移对发育、免疫反应、组织重塑和伤口愈合至关重要。整合素介导的细胞迁移也有助于许多疾病状态，是肿瘤转移、炎症和动脉粥样硬化斑块形成所必需的。丝蛋白与整合素尾部的增加关联抑制细胞迁移。申请人假设丝蛋白与整合素β尾部的调节关联介导了不同的整合素功能，并作为细胞迁移的刹车。为了验证这一假设，他的目标是绘制丝蛋白中整合素结合位点，并识别选择性抑制整合素结合的突变。他将使用这些突变体和先前发现的可上调或下调丝蛋白结合的整合素突变体来表征整合素-丝蛋白相互作用对整合素介导的信号转导的影响。然后，他将研究丝蛋白与整合素结合增加后对细胞迁移的抑制是由于观察到的对整合素信号传导的影响，还是由于局部丝蛋白介导的对肌动蛋白丝交联的影响，还是这两种过程的结合。最后，他将评估丝蛋白水解、丝蛋白或整合素磷酸化以及整合素与其他丝蛋白结合蛋白之间的竞争对丝蛋白与整合素β尾部关联的影响。这些研究将描述控制细胞迁移的途径的调节和作用机制。细胞迁移是一个受到严格控制的过程，在许多生物现象中起着核心作用。因此，这些研究将深入了解健康和疾病过程中必不可少的过程，并可能确定治疗癌症，关节炎和动脉粥样硬化的新治疗靶点。