Regulation of Cell Migration by the APC-Microtubule Complex

APC-微管复合物对细胞迁移的调节

• 批准号: 7683847
• 负责人: W. James Nelson
• 金额: $ 28.72万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-25 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683847
• 关键词: Actins; Adenomatous Polyposis Coli; Adhesions; Affect; Astrocytes; Binding; Binding Proteins; Biochemical; Biological Assay; Bundling; Capsid Proteins; Cells; Complex; Cytoskeleton; Development; Disease; Epithelial; Epithelial Cells; F-Actin; Fibroblasts; Fluorescence Resonance Energy Transfer; Gene Expression; Growth Factor; Image; In Vitro; Laboratories; Life; Mediating; Membrane; Microscopy; Microtubules; Modification; Morphogenesis; Neoplasm Metastasis; Neurites; Normal tissue morphology; Pathway interactions; Pattern; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Plus End of the Microtubule; Process; Protein Dynamics; Proteins; RNA Interference; Recruitment Activity; Regulation; Regulatory Pathway; Resolution; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Solutions; To specify; Tumor Suppressor Proteins; beta catenin; cell motility; directional cell; extracellular; migration; mutant; novel; protein complex; protein protein interaction; reconstitution; repaired; response; scaffold; spatiotemporal; tumor

Directional cell migration towards extracellular signals is essential for normal tissue morphogenesis and repair, and abnormalities in the regulatory pathways involved result in severe pathological consequences during tumor metastasis. This proposal focuses on the role of the adenomatous polyposis coli (ARC) multi- protein complex in regulating microtubule and actin cytoskeletons in response to extracellular signals that stimulate cell migration and adhesion. APC was originally discovered as a tumor suppressor protein that controls the level of beta-catenin. APC is a large scaffolding protein with many binding partners that also regulates microtubules and directional cell migration in response to extracellular signals. Although the importance of APC in diverse morphological processes is well established, very little is known about how extracellular signals affect its interaction with cytoskeletal binding partners and regulation of microtubule dynamics, or how the APC scaffolding complex influences membrane dynamics and cell migration Our hypothesis is that APC coordinates microtubule and actin reorganization by acting as scaffold for cytoskeletal regulators, and that in direct response to extracellular signaling components of the APC scaffold are locally recruited and activated to specify directional membrane extension. We propose a rigorous biochemical analysis of APC regulation and functions combined with novel assays to reconstitute APC- microtubule dynamics in vitro and to image with high spatiotemporal resolution APC-microtubule dynamics in cells. We propose to: 1). Define mechanisms involved in extracellular signal-induced modifications of APC multi-protein complex composition and functions; 2). Dissect and reconstitute effects of APC multi-protein complex components on microtubule dynamics and F-actin binding in vitro; and 3). Characterize APC- mediated localized changes in microtubule and F-actin dynamics in response to extracellular signals. The significance of these proposed studies is that they will define protein-protein interactions in the APC complex that locally regulate cytoskeleton reorganization during cell extension and contact formation, and how the function of this complex is regulated by signaling pathways important in development and disease.

细胞向细胞外信号定向迁移对于正常组织形态发生和 修复以及相关调节途径的异常会导致严重的病理后果 肿瘤转移期间。该提案重点关注腺瘤性结肠息肉病 (ARC) 的多重作用 响应细胞外信号调节微管和肌动蛋白细胞骨架的蛋白质复合物 刺激细胞迁移和粘附。 APC 最初被发现是一种肿瘤抑制蛋白 控制β-连环蛋白的水平。 APC 是一种大型支架蛋白，具有许多结合伙伴，也 响应细胞外信号调节微管和定向细胞迁移。虽然 APC 在不同形态过程中的重要性已广为人知，但对其如何发挥作用却知之甚少。 细胞外信号影响其与细胞骨架结合伙伴的相互作用以及微管的调节 动力学，或 APC 支架复合物如何影响膜动力学和细胞迁移 我们的假设是 APC 通过充当支架来协调微管和肌动蛋白重组 细胞骨架调节剂，以及直接响应 APC 支架的细胞外信号传导成分 局部招募并激活以指定定向膜延伸。我们提出严格的 APC 调节和功能的生化分析与重构 APC-的新测定法相结合 体外微管动力学和高时空分辨率 APC 微管动力学成像 细胞。我们建议：1）。定义细胞外信号诱导的 APC 修饰所涉及的机制 多蛋白复合物的组成和功能； 2）。剖析和重建 APC 多蛋白的作用 复杂成分对微管动力学和 F-肌动蛋白体外结合的影响；和3）。表征 APC- 介导响应细胞外信号的微管和肌动蛋白动力学的局部变化。 这些拟议研究的意义在于它们将定义 APC 中的蛋白质-蛋白质相互作用 在细胞延伸和接触形成过程中局部调节细胞骨架重组的复合物，以及 该复合物的功能如何受到发育和疾病中重要的信号通路的调节。