Minority Predoctoral Fellowship Program

少数族裔博士前奖学金计划

• 批准号: 6984195
• 负责人: DEREK A APPLEWHITE
• 金额: $ 2.87万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984195
• 关键词: actins; binding proteins; cell morphology; cell motility; cytoskeletal proteins; cytoskeleton; fluorescence microscopy; fluorescence recovery after photobleaching; light microscopy; microfilaments; molecular /cellular imaging; predoctoral investigator; protein structure function; protoplasm motility; small interfering RNA; tissue /cell culture

DESCRIPTION (provided by applicant): The actin cytoskeleton is essential for cell motility and depends on the formation of two major actin-based protrusive structures: filopodia and lamellipodia. Both protrusive structures often exist at the leading edge of migrating cells with their formation dependent on the regulation of the barbed-end (fast growing) of actin filaments; however they contribute differentially to cell behavior. Understanding how the cell "determines" which protrusive structures prevails at the leading edge is important to defining the cell's overall migratory behavior and has implication metastasis, development and wound closure. The proposed study focuses on two regulators of the lamellipodia/filopodia transition that bind to the barbed-end of actin filaments, the heterodimeric capping protein (CP) and the Ena/VASP family of molecules. In the proposed study we will: (1) characterize how the various domains of Ena/VASP proteins contribute to filopodia formation using fluorescent and light microscopy, (2) further elucidate CP dynamics using live cell imaging and photobleaching and activation, (3) and investigate how other barbed-end binding proteins contribute to cell motility through depletion by siRNA.

描述（由申请人提供）：肌动蛋白细胞骨架对细胞运动至关重要，并取决于两种主要肌动蛋白基膨胀结构的形成：丝状伪足和板状伪足。这两种突起结构通常存在于迁移细胞的前缘，它们的形成依赖于肌动蛋白丝的倒刺末端（快速生长）的调节;然而，它们对细胞行为的贡献不同。了解细胞如何“确定”前沿存在哪些突起结构对于定义细胞的整体迁移行为非常重要，并涉及转移、发育和伤口闭合。拟议的研究集中在两个监管机构的片状伪足/丝状伪足过渡，结合到肌动蛋白丝，异二聚体帽蛋白（CP）和Ena/VASP家族的分子的倒钩端。在拟议的研究中，我们将：（1）使用荧光和光学显微镜来表征Ena/VASP蛋白的各个结构域如何有助于丝状伪足的形成，（2）使用活细胞成像和光漂白和活化来进一步阐明CP动力学，（3）并研究其他有刺末端结合蛋白如何通过siRNA消耗来促进细胞运动。