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Interrogating the role of retriever sorting complex in adipose insulin-sensitivity and GLUT4 recycling

Department of Biology & Biochemistry

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Dr Paul Whitley Applications accepted all year round Self-Funded PhD Students Only

About the Project

The University of Bath is inviting applications from self-funded applicants for the following PhD opportunity:

Dr Paul Whitley, Department of Biology & Biochemistry
Dr Francoise Koumanov, Department for Health

Overview of the project:

Type 2 diabetes results from cells failing to respond to insulin and remove excess glucose from circulation. This studentship aims to investigate endosomal sorting pathways that may lead to insulin resistance and type 2 diabetes. The student will develop a range of biochemical, cell biology, molecular biology, imaging, proteomics and bioinformatics interdisciplinary skills.

Specifically, the project will focus on the role played by the endosomal sorting complexes retromer and retriever in adipogenesis and the in insulin-stimulated GLUT4 trafficking in adipose and skeletal muscle peripheral tissues. GLUT4 traffics through multiple compartments to reach specialised storage vesicles (GSV) from where it can be recruited to the plasma membrane upon insulin stimulation. Trafficking events involving sorting GLUT4 and other cargo proteins from a late endosomal compartment back into the recycling pool are essential for the formation of the GSV. Retromer and the newly discovered retriever complexes play a role in late-endosomal sorting. In this study, we propose to explore the potential role of the retriever complex in GSV formation. We have generated clonal knockout 3T3-L1 cell lines, using CRISPR/Cas9 technology, lacking Vps29 (common to both retromer and retriever complexes), Vps35 (unique to retromer) and Vps35L/C16orf62 (unique to retriever). We have confirmed the knockout status of the clones, performed some preliminary analysis, which suggest that retriever complex component may be important for adipogenesis.

The student will have the task to further characterise the phenotype of the CRISPR/Cas9 knockout cell lines with particular focus on the mechanisms of differentiation of the pre-adipocytes into adipocytes (adipogenesis) and GLUT4 trafficking.

We have set the following research objectives for the PhD studentship:

1. Characterise the adipogenesis phenotype. Preliminary work has identified that Retriever KO cell line has defective differentiation into adipocytes. The aim will be to identify how the retriever complexes affect adipogenesis.

2. Perform a global proteomic screen for retriever dependent cargos in adipocytes. This will complement data sets from other cell types and identify cargoes specific to adipocytes.

3. Determine whether alterations in Retriever/Retromer expression affects GLUT4 expression, subcellular distribution and recycling.

4. Investigate whether in cellular models of insulin resistance Retriever expression is altered.

Candidate requirements:

Candidates are expected to hold (or be about to obtain) a minimum upper second-class honours degree (or equivalent) in a related subject. Candidates with experience in any molecular and cellular techniques or with an interest in diabetes and metabolic diseases are encouraged to apply.

How to apply:

Informal enquiries are welcomed and should be addressed to Dr Paul Whitley.

Formal applications should be made via the University of Bath’s online application form for a PhD in Biochemistry:

Please ensure that you quote the supervisor’s name and project title in the ‘Your research interests’ section.

More information about applying for a PhD at Bath may be found here:

Funding Notes

We welcome all-year-round applications for students who can fund their own studies or can source their own external scholarship.
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