Coordination of the actin and microtubule cytoskeletons

肌动蛋白和微管细胞骨架的协调

• 批准号: 8233885
• 负责人: JEFF GELLES
• 金额: $ 32.16万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8233885
• 关键词: Actins; Acute Lymphocytic Leukemia; Adenomatous Polyposis Coli; Adenomatous Polyposis Coli Protein; Area; Automobile Driving; Behavior; Binding; Binding Proteins; Binding Sites; Biochemical; Biochemistry; Biological; Biological Process; Biophysics; Cancer Etiology; Cell Shape; Cell physiology; Cells; Cellular Morphology; Cellular biology; Cessation of life; Characteristics; Colorectal Cancer; Complex; Cytoskeleton; Defect; Development; Filament; Fluorescence; G Actin; Gap Junctions; Gene Fusion; Genes; Genetic; Goals; Grant; Human; In Vitro; Individual; Inherited; Investigation; Lead; Life; Link; MLL gene; Malignant Neoplasms; Mediating; Medical; Methods; Microfilaments; Microscopy; Microtubules; Molecular; Morphogenesis; Mutation; Pathway interactions; Polymers; Positioning Attribute; Process; Proteins; Regulation; Regulatory Pathway; Research; Research Personnel; System; Techniques; Testing; Triad Acrylic Resin; Tumor Suppressor Proteins; Ursidae Family; Work; biological systems; cell growth regulation; cell motility; experience; in vivo; insight; light microscopy; novel; polymerization; protein complex; research study; single molecule; tumorigenesis

DESCRIPTION (provided by applicant): The goal of this research is to determine how the dynamics of the actin and microtubule cytoskeletons are coordinated by a group of three interacting mammalian proteins: APC, Dia1, and EB1. This work will define the functions and mechanisms of these proteins, and provide a deeper understanding of the activities and interactions that underlie such processes as cell migration and morphogenesis. This project uses bulk biochemical experiments combined with a novel multi-wavelength single molecule biophysics method tailored to elucidate the mechanisms of complex, multi-component regulatory systems in vitro. In addition, the mechanisms deduced from the experiments in vitro will be tested in vivo to verify that they are important for relevant biological functions of these proteins in living cells (e.g., directed cell migration). The Specific Aims are: (1) Test two key hypotheses about the mechanism by which Dia1 and APC synergize in promoting actin assembly, involving formation of proposed physical complexes among components; (2) Test the additional hypothesis that Dia1 and APC synergize to stimulate actin assembly by a "rocket launcher" mechanism; and (3) Define the mechanisms by which EB1 alone and EB1 plus microtubules regulate and/or organize APC/Dia-induced actin assembly. PUBLIC HEALTH RELEVANCE: This grant is an investigation of the organization and regulation of actin and microtubule cytoskeletons by the combined effects of three proteins: APC, Dia1, and EB1. APC (Adenomatous polyposis coli) is a tumor suppressor. Mutation of the human Apc gene is considered to be an early step in the progression of over 80% of colorectal cancers, inherited and sporadic. In addition, fusion of the EB1 and MLL genes can lead to acute lymphoblastic leukemia. We anticipate that this research will provide new insights into the underlying mechanisms of tumorigenesis.

描述（由申请人提供）：这项研究的目的是确定肌动蛋白和微管细胞骨架的动力学如何由三种相互作用的哺乳动物蛋白组成的组协调：APC，DIA1和EB1。这项工作将定义这些蛋白质的功能和机制，并对诸如细胞迁移和形态发生等过程的活动和相互作用有更深入的了解。该项目使用散装生化实验，结合了一种新型的多波长单分子生物物理学方法，该方法量身定制，以阐明体外复杂的多组分调节系统的机制。此外，将在体外测试实验中从实验中得出的机制，以验证它们对于这些蛋白质在活细胞中这些蛋白的相关生物学功能很重要（例如，定向细胞迁移）。具体目的是：（1）测试DIA1和APC在促进肌动蛋白组装方面协同作用的两个关键假设，涉及组件之间提出的物理复合物的形成； （2）检验DIA1和APC协同刺激肌动蛋白组装的其他假设，通过“火箭发射器”机制刺激肌动蛋白组件； （3）定义单独EB1和EB1加微管调节和/或组织APC/DIAS诱导的肌动蛋白组件的机制。 公共卫生相关性：这笔赠款是对肌动蛋白和微管细胞骨架的组织和调节的调查，该组织通过三种蛋白质的综合作用：APC，DIA1和EB1。 APC（腺瘤息肉大肠杆菌）是肿瘤抑制剂。人类APC基因的突变被认为是超过80％的结直肠癌（遗传和零星）进展的早期步骤。另外，EB1和MLL基因的融合会导致急性淋巴细胞白血病。我们预计这项研究将为肿瘤发生的潜在机制提供新的见解。