Studying the role of Aurora A and Protein Phosphatase 6 in regulation of NDC80 during mitosis

研究 Aurora A 和蛋白磷酸酶 6 在有丝分裂过程中 NDC80 调节中的作用

• 批准号: 2605490
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2605490
• 关键词: Studying; role; Aurora; Protein; Phosphatase

Accurate chromosome segregation is crucial for the generation of new cells and tissues. Aurora kinases and their regulators play a key role in accurate chromosome segregation through regulation of microtubule dynamics and microtubule binding to kinetochores during mitotic spindle formation. However, many of the mechanistic details remain unclear. We have recently found that Aurora A is the primary regulator of the microtubule-kinetochore interface through phosphorylation of the microtubule binding kinetochore protein NDC80. Aurora A activity is counteracted by protein phosphatase 6 (PP6), and cells lacking PP6 show elevated NDC80 phosphorylation and chromosome segregation defects.In this project we plan to study the role of the Aurora A-PP6 axis and Aurora B in regulating the microtubule cytoskeleton during cell division. Our hypothesis is that the Aurora A pathway promotes microtubule attachments to kinetochores and that Aurora B acts as the sensor for these defects through its role in the spindle assembly checkpoint. Our previous work has focussed on the role of the Aurora A-PP6 pathway in human cancer cells and has generated many of the reagents and tools required. This new project will take advantage of genome engineering, microscopy and optical methods to manipulate NDC80 during cell division to better understand the mechanism of Aurora A regulation. A cornerstone of our approach is quantitative imaging and single molecule imaging of protein dynamics, and computational modelling of the dynamics of the cell cycle system. Using these approaches, we aim to describe the dynamics and function of the cell division machinery and roles of Aurora kinases. The DPhil will provide extensive training in:1. Cell-cycle biology and in vitro biochemistry2. Super-resolution microscopy3. Optical methods and chemical/light-activated degron technology4. Genome engineering using CRISPR/CAS95. Computational modelling

准确的染色体分离对于新细胞和组织的产生至关重要。极光激酶及其调节因子通过调节有丝分裂纺锤体形成过程中微管动力学和微管与动粒的结合，在染色体的精确分离中发挥关键作用。然而，许多机械细节仍然不清楚。我们最近发现，极光A是通过磷酸化微管结合动粒蛋白NDC 80的微管-动粒界面的主要调节剂。蛋白磷酸酶6（PP 6）可抑制Aurora A的活性，缺失PP 6的细胞表现出NDC 80磷酸化水平升高和染色体分离缺陷，本课题拟研究Aurora A-PP 6轴和Aurora B在细胞分裂过程中调控微管骨架的作用。我们的假设是，极光A途径促进微管附件的着丝粒和极光B作为这些缺陷的传感器，通过其在纺锤体组装检查点的作用。我们以前的工作集中在Aurora A-PP 6通路在人类癌细胞中的作用，并产生了许多所需的试剂和工具。这个新项目将利用基因组工程、显微镜和光学方法在细胞分裂过程中操纵NDC 80，以更好地了解Aurora A的调控机制。我们方法的基石是蛋白质动力学的定量成像和单分子成像，以及细胞周期系统动力学的计算建模。使用这些方法，我们的目标是描述的动力学和功能的细胞分裂机制和极光激酶的作用。哲学博士将提供广泛的培训：1。细胞周期生物学和体外生物化学2.超分辨率显微镜3.光学方法和化学/光激活降解决定子技术4.使用CRISPR/CAS95的基因组工程。计算建模