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

• 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
• 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

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、极化和迁移。不同亚细胞 站点可以充当 MTOC 以适应这些不同的流程，在这里我们寻求 了解开发过程中如何建立不同的 MTOC 位置。为了分裂，动物细胞使用 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。在差异化 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 这些细胞。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？微管重组对于正常情况下调节细胞分裂和形状至关重要 发育和癌症。中心体缺陷是许多癌症的典型特征[1]，最近的研究 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入侵行为[11,12]。尽管它很重要，但人们对如何做到这一点却知之甚少。 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相协调。细胞周期是一个 严格控制的过程，使用一系列检查点来确保细胞为分裂做好准备。这里 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 线虫确定细胞周期和 MTOC 位置如何协调。我们实验室最近优化了 靶向蛋白质降解的新策略。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 MTOC 位置。目标 2 将使用互补的遗传和生化方法来识别其中哪些 蛋白质将微管固定在特定位置。在目标 3 中，基因筛查将发现新的调控因子 MTOC 位置和细胞周期的协调。由于大多数癌症起源于上皮细胞，我们预计 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。