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The role of protein palmitoylation during mitosis


Institute of Systems, Molecular and Integrative Biology

About the Project

This opportunity will remain open until a suitable candidate has been found, and so early applications are encouraged.

Mitosis is the highly controlled division of cells to produce two new daughters. It is responsible for most of the growth that occurs in humans and errors in mitosis can lead to uncontrolled cell proliferation and cancer. Understanding how mitosis is controlled is therefore a fundamental goal of cell biology. There are many layers of regulation involved in ensuring the fidelity of cell division and it seems likely that even more are yet to be discovered. This project will investigate the role of protein palmitoylation in mitosis in mammalian cells. Palmitoylation is a common (~10% of all mammalian proteins can be palmitoylated) reversible fatty acid modification on specific cysteine residues that typically modulates membrane and protein interactions, protein localisation and/or protein stability [1]. Patterns of mitosis specific protein palmitoylation have previously been observed [2] however very few protein targets have been identified and the function of palmitoylation during mitosis remains a mystery. We have plasmids encoding all 23 human DHHC palmitoyltransferase enzymes [3] and, in the initial part of the project, the student will assess the localisation of these during mitosis to identify the subset of enzymes that target to mitosis specific cellular structures. Candidate enzymes will then be mutated to destroy transferase activity and the impact on mitosis assessed. Using click-chemistry, the student will attempt to label target, palmitoylated, proteins in chemically synchronised mammalian cells using Biotin-PEG-Alkyne reagents. Tagged proteins will be purified by streptavidin affinity chromatography and identified by LC MS/MS analysis. Genuine, mitosis specific, palmitoylation will be verified using protein specific antibodies in mitotic cells and the function of these targets investigated in further detail to reveal insights into their roles during cell division. 

For any enquiries or to show an expression of interest please contact Dr Lee Haynes on with a CV.


Funding Notes

This is a self funded opportunity.

References

1. Zmuda, F. and L.H. Chamberlain, Regulatory effects of post-translational modifications on zDHHC S-acyltransferases. J Biol Chem, 2020. 295(43): p. 14640-14652.
2. Hannoush, R.N. and N. Arenas-Ramirez, Imaging the lipidome: omega-alkynyl fatty acids for detection and cellular visualization of lipid-modified proteins. ACS Chem Biol, 2009. 4(7): p. 581-7.
3. Lemonidis, K., et al., The zDHHC family of S-acyltransferases. Biochem Soc Trans, 2015. 43(2): p. 217-21.

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