Work Package 2.
Molecular regulation of endo-lysosomal processes and cargo identification.
On entering the endo-lysosomal network newly endocytosed integral membrane proteins and their associated proteins and lipids (together termed ‘cargos’), are subjected to one of two fate decisions: either they are sorted into the lysosome for degradation or they are retrieved from this fate and promoted for recycling back to the cell surface, the biosynthetic pathway or other specific organelles. A number of ancient and highly conserved protein assemblies serve to orchestrate these fate decisions (e.g. retromer, retriever, ESCPE-1), but their molecular regulation remain unclear. The aim is to establish the molecular details that define the relationship between degradative versus recycling fate decisions of internalized cell surface transporters and identify the proteins and cargos that play essential roles in nutrient uptake and metabolism.
PhD projects in this Work Package:
PhD student 1 – Biochemical analysis of the assembly of multi-protein endosomal cargo sorting complexesUniversity of Bristol, UK Pete Cullen, pete.cullen[at]bristol.ac.uk
The PhD student will apply and implement recombinant protein technology to isolate and better understand the assembly of large protein complexes in endosomal cargo transport (including cryo-EM analysis). These studies will provide a detailed molecular understanding of the assembly of retriever and the CCC complex, and this knowledge will be applied to the mechanistic analysis of retriever mediated LDLR sorting.
PhD student 2 – Functional analysis of retriever and the CCC complex in endosomal cargo sorting in hepatocytesUniversity of Bristol, UK Pete Cullen, pete.cullen[at]bristol.ac.uk
The PhD student will use quantitative organelle-restricted proteomics to define global cell surface, Golgi apparatus and lysosomal proteomes of genetically modified hepatoma cells. They will establish interactomes for retriever and CCC components. These results will provide a global and unbiased analysis of the role played by the retriever-CCC-WASH endosomal cargo sorting pathway in regulating organellar proteomes in liver cells and how this relates to the control of liver metabolism.
3. Understanding the fate of cargo via ubiquitination by E3 ligases and its role in hepatic function
PhD student 3 – Understanding the fate of cargo via ubiquitination by E3 ligases and its role in hepatic functionAstraZeneca, Cambridge, UK Helen Boyd, helen.boyd[at]strazeneca.com Kevin Moreau, kevin.moreau[at]atrazeneca.com
You are employed and the research is done at AstraZeneca. In addition, you are registered at the University of Amsterdam (AMC), the Netherlands where you will do the thesis defence.
The PhD student will investigate the role of sorting proteins in hepatic function and identify the molecular mechanisms that induce specific stimulated degradation of endo-lysosomal proteins. The student will develop technology for down regulation of receptors and transporters via targeted protein degradation and investigate the expression profile of E3 ligases involved in glucose and lipid metabolism.