Regulating the Coordination of Microtubule Organization and Cell Cycle State

调节微管组织和细胞周期状态的协调

基本信息

批准号：
9403413
负责人：
Maria Danielle Sallee
金额：
$ 0.24万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-12-01 至 2018-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9403413
关键词：
Alpha Cell Animal Model Animals Behavior Biochemical Biological Biological Assay Caenorhabditis elegans Cancer Biology Carcinoma Cell Adhesion Cell Cycle Cell Cycle Stage Cell Differentiation process Cell Shape Cell division Cell physiology Cells Centrosome Coupled Defect Development Disease Ensure Epithelial Cells Event Genes Genetic Genetic Models Genetic Screening Goals Golgi Apparatus Growth Human Image Intestines Investigation Lead Location Malignant Neoplasms Membrane Methods Microtubule-Organizing Center Microtubules Mitosis Mitotic Mitotic spindle Molecular Molecular Analysis Muscle Cells Nuclear Envelope Organism Orthologous Gene Polymers Process Proteins Recruitment Activity Regulation Role Series Shapes Site Structure Study models Surface Testing Visual apical membrane cancer biomarkers cdc Genes daughter cell experimental study genetic approach genome editing insight migration mutant new therapeutic target novel protein degradation screening therapeutic target tool

项目摘要

Project Summary Microtubule organizing centers (MTOCs) generate specific arrangements of microtubules that are essential for many cellular functions, including cell division, polarization, and migration. Different subcellular sites can function as the MTOC in order to accommodate these different processes, and here we seek to understand how different MTOC locations are established during development. To divide, animal cells use centrosomes as MTOCs, localizing specific proteins to grow and anchor microtubules in order to build the mitotic spindle; the microtubules then promote the correct formation of two daughter cells. In differentiating cells, like some epithelial cells, the location of the MTOC changes: microtubule-organizing proteins leave the centrosome and move to a membrane surface, an “MTOC switch” establishing the membrane as the MTOC. Epithelial cells have distinctive shapes and stick together tightly to form cellular sheets, and the microtubule configuration generated by the membrane MTOC is important for the structure and function of these cells. However, this arrangement of microtubules does not permit division, raising an important question: when an epithelial cell divides, how does it change the location of its MTOC to reorganize its microtubules into the mitotic spindle? Microtubule reorganization is critical for regulating cell division and shape both in normal development and in cancer. Centrosomal defects are classic hallmarks of many cancers [1], and recent studies have shown that increasing the microtubule organizing activity of the centrosome causes epithelial cells to divide more readily and display invasive behavior [11,12]. Despite its importance, little is known about how cells control their microtubule organization and the consequent effect on cell division and shape. The goal of this proposal is to understand how epithelial cells establish specific MTOC locations for different functions and how the change in MTOC location is coordinated with cell division. The cell cycle is a tightly controlled process that uses a series of checkpoints to ensure that a cell is prepared for division. Here we propose a series of genetic and biochemical experiments in intestinal epithelial cells in the model organism C. elegans to determine how the cell cycle and MTOC location are coordinated. Our lab has recently optimized a new strategy for targeted protein degradation. This method, coupled with recent methods for genome editing, will allow us to uncover the molecular connection between cell cycle regulators and microtubule organization. Aim 1 will use both new and classic genetic approaches to determine which cell cycle genes regulate the MTOC location. Aim 2 will use complementary genetic and biochemical methods to identify which of these proteins tether microtubules to specific locations. In Aim 3, a genetic screen will uncover new regulators of the coordination of MTOC location and cell cycle. As the majority of cancers are epithelial in origin, we expect that understanding how epithelial cells coordinate their MTOC with cell division will be relevant to cancer biology and may lead to the discovery of useful cancer biomarkers or therapeutic targets.

项目概要微管组织中心（MTOC）产生特定的微管排列，对于许多细胞功能至关重要，包括细胞分裂、极化和不同的亚细胞迁移。站点可以充当 MTOC 以适应这些不同的流程，在这里我们寻求为了了解动物细胞在发育过程中如何建立不同的 MTOC 位置。中心体作为 MTOC，定位特定蛋白质以生长和锚定微管，以构建有丝分裂纺锤体；然后微管促进两个子细胞的正确形成。细胞，像一些上皮细胞一样，MTOC 的位置发生变化：微管组织蛋白离开中心体并移动到膜表面，“MTOC 开关”将膜确立为 MTOC。上皮细胞具有独特的形状并紧密地粘在一起形成细胞片，并且膜MTOC产生的微管构型对于微管的结构和功能很重要然而，这种微管排列不允许分裂，这就提出了一个重要的问题：当上皮细胞分裂时，它如何改变其MTOC的位置以将其微管重组为有丝分裂纺锤体的重组对于正常细胞分裂和形状的调节至关重要中心体缺陷是许多癌症的典型特征[1]，最近的研究表明。研究表明，增加中心体的微管组织活性会导致上皮细胞更容易分裂并表现出侵入性行为[11,12]尽管它很重要，但人们对其如何进行知之甚少。细胞控制其微管组织以及对细胞分裂和形状的后续影响。该提案的目标是了解上皮细胞如何建立特定的 MTOC 位置不同的功能以及 MTOC 位置的变化如何与细胞分裂协调。严格控制的过程，使用一系列检查点来确保细胞准备分裂。我们提出了在模型生物体的肠上皮细胞中进行一系列遗传和生化实验我们的实验室最近对线虫进行了优化，以确定细胞周期和 MTOC 位置的协调方式。这种方法与最近的基因组编辑方法相结合，将使我们能够揭示细胞周期调节因子和微管组织之间的分子联系。目标 1 将使用新的和经典的遗传方法来确定哪些细胞周期基因调节目标 2 将使用互补的遗传和生化方法来识别其中的哪一个。在目标 3 中，蛋白质将微管束缚在特定位置，基因筛选将发现新的调节因子。由于大多数癌症起源于上皮细胞，因此我们预计 MTOC 位置和细胞周期的协调。了解上皮细胞如何协调 MTOC 与细胞分裂将与癌症生物学相关并可能导致有用的癌症生物标志物或治疗靶点的发现。