Undergraduate Research: Coordinated Cytoskeletal Dynamics and Membrane Remodeling in Cellular Shape Change

本科生研究：细胞形状变化中的协调细胞骨架动力学和膜重塑

基本信息

批准号：
10586656
负责人：
Stephanie Gupton
金额：
$ 0.95万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10586656
关键词：
Affect Area Axon Biochemical Biological Models Cell Shape Cell membrane Cells Characteristics Clathrin Complex Computer Vision Systems Computer software Cues Cytoskeleton Data Detection Development Diffusion Distal Endocytosis Environment Event Excision Exocytosis Frequencies Goals Growth Cones Heterogeneity In Vitro Lateral Machine Learning Malignant Neoplasms Mediating Melanoma Cell Membrane Modeling Molecular Monomeric GTP-Binding Proteins Morphogenesis Morphology Neoplasm Metastasis Nervous system structure Neurites Neuronal Plasticity Neurons PTK2 gene Pathogenesis Pathologic Pathology Pathway interactions Phenotype Phosphorylation Phosphotransferases Physiology Plasma Play Population Program Description Protein Tyrosine Kinase Proteins Publishing Regulation of Exocytosis Research Resistance Role Running SNAP receptor Shapes Stretching Surface Synapses System Techniques Testing Tetanus Ubiquitination Vesicle Work active control cell motility cell type extracellular in vivo melanoma morphogens neural network neuron development novel physical model programs response synaptogenesis trafficking tumor progression ubiquitin-protein ligase undergraduate research vesicular SNARE proteins

项目摘要

PROGRAM DESCRIPTION The overarching research goal of my lab is to define cellular and molecular mechanisms mediating cellular shape change. Cellular shape change is a fundamental characteristic of metazoan cells, key to development, physiology, and pathology. The formation and plasticity of neural networks are key examples of cell shape change during development and physiology, whereas cell shape and motility goes awry in pathological conditions, such as melanoma. We study two main themes during cellular shape change: The active control of the cytoskeleton, which is acknowledged as critical to cellular shape change, and the concurrent remodeling of the plasma membrane, which is perhaps less well appreciated. Although many cytoskeletal and membrane remodeling components are known and their biochemical and structural characteristics described, we lack a systematic understanding of how these disparate systems are regulated and coordinated to orchestrate cellular shape change. Perhaps the most important problem in cell morphogenesis is understanding how cells perceive cues in their environment and convert this extracellular information into shape changes through coordinated cytoskeletal dynamics and plasma membrane remodeling; this is the focus of this proposal. Functions of small GTPases and kinases have been extensively studied in regulating cytoskeletal dynamics and membrane remodeling. Work from my lab identified an emerging role for E3 ubiquitin ligases in regulated cellular shape change. We identified two E3 ubiquitin ligases, TRIM9 and TRIM67, which regulate cytoskeletal and exocytic proteins and cellular shape changes in response to netrin. The extracellular morphogen netrin promotes neuronal morphogenesis and cancer progression. Despite these important consequences, we know little about how cells interpret netrin into shape changes. TRIM9 and TRIM67 provide an excellent opportunity to investigate the function of ubiquitination in cytoskeletal and membrane remodeling, and how these functions are coordinated during netrin triggered cell shape change and motility. TRIM9 and TRIM67 share similar sequences, localization, and interaction partners, however our studies identified distinct functions of these related proteins and antagonistic phenotypes associated with their deletion. The overarching goal of this program is to test the hypothesis that TRIM9 and TRIM67 coordinate cytoskeletal dynamics and exocytosis during netrin-dependent morphogenesis in multiple cell types. Since netrin plays roles in both neuronal development and cancer pathogenesis, our work will exploit developing neurons and migrating melanoma cells as model systems. Our preliminary and published data indicate both cell types respond to netrin and express TRIM9 and TRIM67. Our work will illuminate fundamental generalities and cell type specific mechanisms of shape change, providing mechanistic understanding of the coordination of the cytoskeleton and membrane trafficking during development and metastasis.

项目描述我实验室的首要研究目标是定义介导细胞形状的细胞和分子机制改变。细胞形状变化是后生动物细胞的基本特征，是发育的关键，生理学和病理学。神经网络的形成和可塑性是细胞形状的关键例子在发育和生理过程中会发生变化，而细胞形状和运动在病理过程中会出现问题条件，例如黑色素瘤。我们研究细胞形状变化过程中的两个主要主题：细胞骨架被认为对细胞形状的变化以及同时的重塑至关重要质膜，这可能不太受重视。尽管许多细胞骨架和膜重塑成分是已知的，并且它们的生化和结构特征已被描述，我们缺乏系统地了解这些不同的系统如何调节和协调以协调细胞形状改变。也许细胞形态发生中最重要的问题是了解细胞如何感知环境中的线索，并通过协调将这些细胞外信息转化为形状变化细胞骨架动力学和质膜重塑；这是本提案的重点。小的功能 GTP 酶和激酶在调节细胞骨架动力学和膜方面已得到广泛研究重塑。我实验室的工作确定了 E3 泛素连接酶在调节细胞形状中的新兴作用改变。我们鉴定了两种 E3 泛素连接酶 TRIM9 和 TRIM67，它们调节细胞骨架和胞吐蛋白质和细胞形状会因 netrin 的反应而发生变化。细胞外形态发生素 netrin 促进神经元形态发生和癌症进展。尽管有这些重要的后果，我们对细胞如何将 netrin 解释为形状变化。 TRIM9 和 TRIM67 提供了一个绝佳的机会来调查泛素化在细胞骨架和膜重塑中的功能，以及这些功能如何协调在 netrin 触发细胞形状变化和运动期间。 TRIM9 和 TRIM67 具有相似的序列、定位、和相互作用伙伴，然而我们的研究发现了这些相关蛋白质的不同功能与其缺失相关的拮抗表型。该计划的总体目标是测试假设 TRIM9 和 TRIM67 在 netrin 依赖性过程中协调细胞骨架动力学和胞吐作用多种细胞类型的形态发生。由于 netrin 在神经元发育和癌症中发挥作用发病机制，我们的工作将利用发育中的神经元和迁移的黑色素瘤细胞作为模型系统。我们的初步和已发表的数据表明两种细胞类型均对 netrin 做出反应并表达 TRIM9 和 TRIM67。我们的工作将阐明形状变化的基本共性和细胞类型特定机制，提供对发育过程中细胞骨架和膜运输协调的机制理解和转移。