Kinase regulation of COPII transport in neuronal differentiation

The challenges of the developing human brain include co-ordinating the birth and migration of 100 billion neurons and the formation of their 1000 trillion synaptic connections. Protein phosphorylation by kinase enzymes is one of the mechanisms that orchestrates these processes. The ubiquitous and highly conserved C-Src tyrosine kinase is enriched in the brain and there is neuronal-specific splicing of C-Src to yield two further isoforms, N1-Src and N2-Src. We and others have shown that the neuronal Srcs play a role in both neurogenesis and neuronal differentiation during development. In a recent screen, we discovered that N2-Src interacts with proteins of the COPII complex, which coats secretory vesicles that traffic from the ER to the Golgi apparatus. Furthermore, overexpression of a COPII protein disrupts N2-Src-dependent differentiation in a neuronal cell line. However, it is unclear how N2-Src-dependent regulation of COPII transport is involved in neuronal differentiation. This project will draw on expertise in kinase signalling and membrane trafficking in the Evans and Pryor labs to discover how N2-Src regulates the COPII complex. Mapping the sites on COPII proteins that are phosphorylated by N2-Src will allow the generation of phosphomutants that can be used to probe their function in COPII transport in vitro and in a cell-based model of neuronal differentiation. Training will be provided in a wide range of biochemical and cell biological approaches, with a particular focus on proteomics and fluorescence imaging. The data arising from this project will inform fundamental mechanisms of neuronal development, but will also be applicable to optimising the differentiation of neuronal stem cells in the lab and improving differentiating therapies in diseases such as neuroblastoma.

Relevant publications:
Lewis PA et al. (2017) J Neurosci. 37(35):8477-8485
Keenan S et al. (2017) Sci. Rep. 7, 43106
Keenan S et al. (2015) FEBS Lett, 589(15), 1995-2000

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