Investigating the role of the clathrin light chain in vivo

Clathrin-mediated endocytosis (CME) is the process by which proteins are transported into cells from their surfaces. This process is fundamental for organism development and adult tissue homeostasis, and occurs via the invagination of the plasma membrane which pinches off to form cargo-containing vesicles. The clathrin light chain (CLC) is a component of the molecular lattice that facilitates membrane bending on the inner surface of the cell. Based on the evidence from in vitro and cellular work, we hypothesise that CLCs are required when membrane properties are refractory to bending i.e. high tension, and/or for the uptake of specific cargos such as large receptors. This project investigates the role of CLCs in the developing tissues of Drosophila melanogaster – in an environment containing native biomechanical cues. Together, the data from this project suggests that there is a differential requirement for CLC at the tissue, cellular and protein level in Drosophila melanogaster. Knockdown of CLC in the developing retina interfered with the morphogenesis of some tissues but not others, and perturbed the morphology of some cell types within the retina, while other cell types remained unaffected. Additionally, an enhancer suppressor screen has identified candidates that genetically interact with CLC and has potentially highlighted signalling pathways/proteins that are dependent on CLC during development. This context-dependent requirement for CLC aligns with the current literature and demonstrates that investigating endocytic dynamics in a living organism is key to building a holistic understanding of this process.