Regulation of Cell Migration by the APC-Microtubule Complex

APC-微管复合物对细胞迁移的调节

基本信息

批准号：
7290967
负责人：
W. James Nelson
金额：
$ 28.18万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
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/7290967
关键词：
Actins Adenomatous Polyposis Coli Adhesions Affect Astrocytes Binding Binding Proteins Biochemical Biological Assay Bundling Cadherins Capsid Proteins Cell-Cell Adhesion Cells Complex Cytoskeleton Development Disease Epithelial Epithelial Cells Extracellular Matrix F-Actin Fibroblasts Fluorescence Gene Expression Growth Factor Image In Vitro Laboratories Lasers Localized Mediating Membrane Microtubules Modification Morphogenesis Neoplasm Metastasis Nerve Growth Factor 1 Nerve Growth Factor Pathway Neurites Normal tissue morphology Pathway interactions Pattern Personal Satisfaction Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Photobleaching Plus End of the Microtubule Process Proteins RNA Interference Recruitment Activity Regulation Regulatory Pathway Resolution Role Scaffolding Protein Signal Pathway Signal Transduction Solutions To specify Tumor Suppressor Proteins Variant beta catenin cell motility directional cell extracellular migration mutant novel protein function protein protein interaction reconstitution repaired response scaffold spatiotemporal tumor

项目摘要

DESCRIPTION (provided by applicant): 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 (APC) 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.

描述（由申请人提供）：细胞向细胞外信号的定向迁移对于正常组织形态发生和修复至关重要，并且所涉及的调节途径的异常会在肿瘤转移过程中导致严重的病理后果。该提案重点关注腺瘤性大肠杆菌（APC）多蛋白复合物在响应刺激细胞迁移和粘附的细胞外信号而调节微管和肌动蛋白细胞骨架中的作用。 APC 最初被发现是一种控制 β-连环蛋白水平的肿瘤抑制蛋白。 APC 是一种大型支架蛋白，具有许多结合伙伴，还可以响应细胞外信号调节微管和定向细胞迁移。尽管 APC 在不同形态过程中的重要性已得到充分证实，但关于细胞外信号如何影响其与细胞骨架结合伴侣的相互作用和微管动力学的调节，或者 APC 支架复合物如何影响膜动力学和细胞迁移，我们知之甚少。 APC 通过充当细胞骨架调节因子的支架来协调微管和肌动蛋白重组，并且直接响应 APC 支架的细胞外信号成分，局部招募和激活以指定方向膜延伸。我们提出对 APC 调节和功能进行严格的生化分析，并结合新的测定方法，在体外重建 APC 微管动力学，并以高时空分辨率对细胞中的 APC 微管动力学进行成像。我们建议：1）。定义细胞外信号诱导的 APC 多蛋白复合物组成和功能修饰的机制； 2）。剖析并重建 APC 多蛋白复合物成分对体外微管动力学和 F-肌动蛋白结合的影响；和3）。表征 APC 介导的微管和 F-肌动蛋白动力学响应细胞外信号的局部变化。这些拟议研究的意义在于，它们将定义 APC 复合物中的蛋白质-蛋白质相互作用，该复合物在细胞延伸和接触形成过程中局部调节细胞骨架重组，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。