Deciphering p53 signaling
Dr Manuel Mueller, Department of Chemistry, King's College London.
Dr Argyris Politis, Department of Chemistry, King's College London.
Dr Sean Devenish, Head of Research and Development, Fluidic Analytics Limited
We are offering a fully-funded studentship for an outstanding PhD candidate who will produce and study PTM variants of p53 to deepen our understanding of the function and regulation of this key tumour suppressor protein.
Cells are continuously exposed to cytotoxic damage. Appropriate responses – ranging from cell cycle arrest all the way to controlled cell death – are a critical line of defence against the onset of cancer. A key regulator of these pathways is the tumour suppressor protein p53, which is frequently mutated in cancer (1). p53 itself is tightly regulated by post-translational modifications (PTMs). Despite their importance, the molecular details of how specific PTMs and their combinations control p53 activity remain elusive. This iCASE PhD project aims to fill this gap through a collaborative chemical biology/biophysics approach in the Müller and Politis Labs at KCL and Dr Sean Devenish at Fluidic Analytics.
The Müller lab has developed a strategy to access site-specifically modified p53 using protein semi-synthesis (manuscript in preparation). During the first year, you will receive training in our cutting-edge semi-synthesis methodology – a combination of chemical peptide synthesis and recombinant technology (for applications, see refs 2,3). In this way, you will prepare a panel of site-specifically modified p53 variants, and further develop semi-synthesis approaches to produce previously inaccessible p53 variants. You will then characterize the biophysical effects of p53 PTMs using innovative approaches developed by Fluidic Analytics (4) and the Politis Lab (5). Large-scale structural and functional changes will be measured through DNA binding assays and diffusive sizing during a 4 months placement at Fluidic Analytics. The impact of PTMs on local structure and dynamics will be determined via Hydrogen-Deuterium Exchange coupled to Mass Spectrometry (Politis lab).
This project will provide unique training to the PhD student in cutting edge chemical biology, structural biology and microfluidics-based analytical biochemistry in academic and industrial settings. Moreover, the results will further our understanding of p53, how it integrates stress signals in the form of PTMs to prevent cancer, and how these mechanisms are distorted by cancer-associated p53 mutations.
- (1) Hafner, A et al. (2019) The multiple mechanisms that regulate p53 activity and cell fate. Nature Reviews Mol. Cell Biol. 20: 199.
- (2) Müller, MM et al., (2016) A two-state activation mechanism controls the histone methyltransferase Suv39h1. Nature Chem Biol. 12: 188.
- (3) Müller, MM and Muir, T. (2015) Histones: At the Crossroads of Peptide and Protein Chemistry. Chemical Reviews 115: 2296.
- (4) Yates et al., (2015) Latent analysis of unmodified biomolecules and their complexes in solution with attomole detection sensitivity. Nature Chemistry, 7: 802.
- (5) Ahdash, Z et al., (2019) HDX-MS reveals nucleotide-dependent, anti-correlated opening and closure of SecA and SecY channels of the bacterial translocon. eLife 8: e47402.
eligibility and application
Applicants must hold, or be expected to achieve, a first or high upper second-class undergraduate honours degree or equivalent (for example BA, BSc, MSci) or a Masters degree in a relevant subject. This project is funded by a 4-year BBSRC studentship, applicants should ensure they have understood the funding eligibility criteria for these studentships. Unfortunately international students are not eligible for programme funding on this project.
Applicants should hold a BSC or Msc in Biochemistry, Biophysics or Chemistry, and have an understanding of protein (bio)chemistry and biophysical methods. Wet-lab experience in protein biochemistry and/or peptide synthesis will be a strong advantage. Candidates must have a fascination for interdisciplinary research, excellent communication skills and work well in a team.
For more information regarding the project, please contact Dr Manuel Mueller
Download the APPLICATION GUIDELINES here.