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

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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