Investigation of the role of post-translational modifications in hypoxia signalling

The hypoxia signalling pathway is required for the adaptation of cells to a low oxygen environment, which cells can experience in various physiological and pathological circumstances related to high altitude, development, wound healing and cancer. Hypoxia inducible factor (HIF) is the main transcription factor involved in the cellular adaptation to hypoxia and it is therefore crucial to understand how HIF and its main partners are regulated. Whilst HIF regulation by hydroxylation has been described for years, no other HIF post-translational modifications (PTMs) triggered by hypoxia has been investigated in an unbiased way, and their impact on binding interactions have not yet been elucidated. We have initiated the elucidation of HIF PTMs using mass spectrometry (MS)-based proteomics strategies. This project will follow on our initial discoveries by investigating the temporal regulation of HIF PTMS and their specificity to the different HIF family members. In addition to understanding the type and site of protein modification as a function of different O2 levels, state-of-the art MS techniques will be exploited to understand how these PTMs occur in combination (at the level of the intact protein). Importantly, the student will have an exciting opportunity of testing the relevance of the major PTMs discovered using live-cell imaging experiments by expressing PTM mimetic and ‘dead’ mutants of HIF, tagged with fluorescent proteins. The imaging approach will be used to test HIF dynamics and binding to partners at single cell and single molecule level. The student will work and will be trained both in the world class Centre for Proteome Research, within a group working at the forefront of protein modifications analysis, and the Centre for Cell Imaging. The project is suited to a student with at least a good B.Sc. Upper Second in Biological or Life Sciences, with a solid knowledge in biochemistry and cell signalling.