Project information
Centrosome Isolation to Locate Interactions Of TTBK2 And Create new Tools to Investigate primary Cilia (CILIOTACTIC)

Primary cilia (PC) are small antenna-like organelles that extend from the centrosome of many cells. PC are key signalling hubs that have essential roles in development and tissue homeostasis. Importantly, it is changes to composition of the centrosome that determine if, and when, PC will form. Critical for the formation of PC is Tau Tubulin Kinase 2 (TTBK2), which is recruited to the mother centriole (MC) of the centrosome to trigger cilium assembly. However, developing a mechanistic understanding of TTBK2 at the MC has been challenging as centrosomes only represent a tiny proportion of the cell and many ciliary proteins, including TTBK2, have roles in other cellular compartments.

To address this challenge, I will leverage a centrosome affinity purification technique to determine how MC-associated TTBK2 governs PC formation and function. Firstly, I will perform discovery proteomics on centrosome isolations to learn how perturbations to TTBK2 affect the molecular composition of the MC and, using cell and molecular biology, investigate the functional consequences of these changes. Secondly, I will combine centrosome isolation with super resolution microscopy to investigate MC structure. Finally, I will adapt centrosome isolation for use in complex systems such as organoids and investigate the potential to isolate an intact primary cilium.

Completing this work as an MSCA fellow will give me the opportunity to apply my expertise to a different biological field and enable me to develop the technical and personal skills necessary to establish myself as an independent scientist. To facilitate project success, I will complete this work in the Laboratory of Cilia and Centrosome Biology at Masaryk University, with the guidance of Asst. Prof. Lukáš Čajánek. Together, this project will help define mechanisms underlying PC-related functions of TTBK2 and expand our understanding of the regulation of these essential sensory organelles and their role in health and disease.