About the Project
The proposed work will directly test the hypothesis, and it will involve a combination of techniques, including those in molecular biology (e.g., cloning and tagging and site-directed mutagenesis), protein chemistry (e.g., glutathione S-transferase/Histidine pulldowns & co-immunoprecipitations followed by western blotting, protein purification and assays for SUMOylation/deSUMOyation), cell biology (e.g., cultures of clonal cell lines and neuronal cells, and DNA & sh/siRNA transfections/CRISPR-Cas9-mediated knockdown/knockout and replacement), together with In-cell quantitative analysis of the aggregation and the inclusion of α-synuclein under different SUMOylation statuses and biochemical examinations of cell viability (e.g., cytochrome c release, caspase activation, and MTT and LDH assays).
Science Graduate School
As a PhD student in one of the science departments at the University of Sheffield, you’ll be part of the Science Graduate School. You’ll get access to training opportunities designed to support your career development by helping you gain professional skills that are essential in all areas of science. You’ll be able to learn how to recognise good research and research behaviour, improve your communication abilities and experience the breadth of technologies that are used in academia, industry and many related careers. Visit http://www.sheffield.ac.uk/sgs to learn more.
All applicants should ensure that both references are uploaded onto their application as a decision will be unable to be made without this information.
1. K. Rosborough, N. Patel, L. V. Kalia, alpha-Synuclein and Parkinsonism: Updates and Future Perspectives. Current neurology and neuroscience reports 17, 31 (2017).
2. P. Krumova et al., Sumoylation inhibits alpha-synuclein aggregation and toxicity. J Cell Biol 194, 49-60 (2011).
3. C. Guo et al., SENP3-mediated deSUMOylation of dynamin-related protein 1 promotes cell death following ischaemia. Embo j 32, 1514-1528 (2013).
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