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.

项目摘要 微管组织中心（MTOCS）生成特定的微管的布置 对于许多细胞功能，包括细胞分裂，极化和迁移至关重要。不同的亚细胞 站点可以作为MTOC起作用以适应这些不同的过程，在这里我们试图 了解在开发过程中如何建立不同的MTOC位置。分裂，动物细胞使用 中心体为mTOC，将特定蛋白质定位以生长和锚定微管以构建 有丝分裂主轴；然后，微管促进了两个子细胞的正确形成。在区分中 细胞，像某些上皮细胞一样，MTOC的位置发生了变化：微管 - 组织蛋白离开 中心体和移动到膜表面，一种“ mTOC开关”，将膜作为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，然后 膜MTOC生成的微管构型对于对的结构和功能很重要 这些细胞。但是，这种微管的安排不允许划分，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的位置将其微管重组为 有丝分裂纺锤？微管重组对于控制细胞分裂和形状至关重要 发育和癌症。中心缺陷是许多癌症的经典标志[1]，也是最近的研究 已经表明，增加中心体的微管组织活性会导致上皮细胞 更容易划分并显示侵入性行为[11,12]。尽管它很重要，但对如何 细胞控制其微管组织，并对细胞分裂和形状产生影响。 该建议的目的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。细胞周期是 严格控制的过程使用一系列检查点来确保为划分准备一个单元格。这里 我们提出了模型生物体中肠上皮细胞中的一系列遗传和生化实验 秀丽隐杆线虫确定细胞周期和MTOC位置的协调方式。我们的实验室最近优化了 靶向蛋白质降解的新策略。这种方法以及最新的基因组编辑方法， 将使我们能够发现细胞周期调节剂与微管组织之间的分子连接。 AIM 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。 AIM 2将使用完整的遗传和生化方法来识别其中哪些 蛋白质将微管链接到特定位置。在AIM 3中，遗传屏幕将发现新的调节器 MTOC位置和细胞周期的协调。由于大多数癌症的起源都是上皮的，我们希望 了解上皮细胞如何与细胞分裂协调其MTOC与癌症生物学有关 并可能导致发现有用的癌症生物标志物或治疗靶标。