ESR3

Dissecting protein networks and regulatory mechanisms driving ciliary disassembly


Cilia disassembly is a crucial process for cells to enter the cell cycle and proliferate but also for the function of certain cell types (i.e. retinal pigment epithelial cells). In contrast to ciliary assembly, disassembly is only partially understood and especially the timing of disassembly induction and execution is unclear. In this study, we aim at identifying the protein networks, involved in ciliary disassembly in a time resolved fashion to understand both the mechanisms of initiation and execution of this process. Therefore, ESR3 will use affinity- and proximity- based methods to quantitatively study the network by analysing the protein complexes of key players and by defining the protein repertoire of cilia at different stages. CRISPR/Cas9 will be used to endogenously tag bait proteins, TurboID as well as FLAG-based affinity enrichment will be used to quantitatively analyse the complexes using high-resolution mass spectrometry and statistical analysis of the quantitative data. ESR3 will complement initial results by alternative proteomic approaches and by localization studies using super-resolution microscopy (gSTED) and live cell imaging. The data will be integrated into the existing ciliary landscape and analysed to define testable hypothesis that will be validated in differentiated retinal pigment epithelial cells.

Partner: Eberhard Karls University Tubingen, Germany

Supervisor: dr K. Boldt