About the Project
The proposed work will involve a combination of techniques, including molecular biology (e.g., cloning, tagging, mutagenesis and CRISPR-Cas9 technology), protein chemistry (e.g., glutathione S-transferase/Histidine pulldowns, western blotting for LC3 conversion and p62 degradation, co-immunoprecipitations, protein purification and assays for SUMOylation/deSUMOyation), cell biology (e.g., cultures of clonal cell lines, neuronal cell lines and primary rat cortical neurons; DNA & sh/siRNA transfections, pexophagy induction by cell stressors such as hypoxia, hydrogen peroxide and clofibrate), imaging detection of pexophagy (e.g., immunocytochemistry, fluorescence, confocal and electron microscopic analysis of morphological changes in peroxisomes as well as “pexophagic flux” in cells expressing a novel biosensor pexo-pHfluorin), and peroxisome function analysis (e.g., ROS production and cell viability examinations upon and following stressed conditions).
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. O. Y. Al-Dirbashi et al., Zellweger syndrome caused by PEX13 deficiency: report of two novel mutations. American journal of medical genetics. Part A 149a, 1219-1223 (2009).
2. C. M. Cipolla, I. J. Lodhi, Peroxisomal Dysfunction in Age-Related Diseases. Trends in endocrinology and metabolism: TEM 28, 297-308 (2017).
3. T. Y. Nazarko, Pexophagy is responsible for 65% of cases of peroxisome biogenesis disorders. Autophagy 13, 991-994 (2017).
4. K. M. Walter et al., Hif-2alpha promotes degradation of mammalian peroxisomes by selective autophagy. Cell metabolism 20, 882-897 (2014).
5. M. van Hagen, R. M. Overmeer, S. S. Abolvardi, A. C. Vertegaal, RNF4 and VHL regulate the proteasomal degradation of SUMO-conjugated Hypoxia-Inducible Factor-2alpha. Nucleic acids research 38, 1922-1931 (2010).
6. C. Huang et al., SENP3 is responsible for HIF-1 transactivation under mild oxidative stress via p300 de-SUMOylation. The EMBO journal 28, 2748-2762 (2009).
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