Understanding the process by which DNA breaks are healed via by non-homologous end joining is vital for developing the design and precision of future anti-cancer medicines and the optimization of CRISPR/Cas9-based gene editing. The X-ray repair cross-complementing protein 4 (XRCC4) is a core DNA repair factor known for its importance in this type of DNA repair. As part of the DNA repair process it interacts with other proteins including the core non-homologous end joining factor XRCC4-like factor (XLF), implicated in tethering the DNA ends together promoting DNA-end ligation, the nucleoskeleton protein intermediate filament family orphan 1 (IFFO1), to stabilise the broken ends of the DNA, and the non-homologous end joining ligating enzyme DNA ligase 4 (LIG4), that facilitates the final ligation step in non-homologous end joining. XRCC4 also interacts with chains of small ubiquitin-like modifier (SUMO) units, which experiments suggest block the sites where the other proteins and enzymes interact with XRCC4. In this way SUMO regulates the action of XRCC4 and the non-homologous end joining of DNA.
This project will utilize Electron Paramagnetic Resonance (EPR) methods combined with the results of other biophysical experiments to characterize the structure of XRCC4 interacting with SUMO. EPR pulsed dipolar spectroscopy (PDS) techniques, such as Double Electron Electron Resonance (DEER) will be used to measure distances between spin-labels added to the surfaces of the proteins at known sites. This will enable us to probe suspected conformation changes induced by SUMO binding, investigate the possibility of SUMO chains crosslinking between different binding sites and test the validity of existing models. Results from the EPR experiments will be combined with existing data to produce models of the interacting complexes using molecular dynamics simulation and protein-protein docking including experimentally derived restraints to guide the calculations.
The research is highly multidisciplinary and will equip the student will a wide range of skills in molecular biology; including cloning, cell culturing, protein expression and purification, EPR spectroscopy measurements and data analysis and computational tools to probe protein-protein interactions. Training will be provided in all areas and thus the project could suit a candidate from either a physical sciences background who is willing to learn biochemical techniques or a biological sciences background who is interested in learning spectroscopy and computational methods.
Interested students should contact the supervisory team for informal discussions prior to making an application for this project.
https://www.research.manchester.ac.uk/portal/alice.bowen.html
https://www.research.manchester.ac.uk/portal/christine.schmidt.html
https://www.research.manchester.ac.uk/portal/sam.hay.html
https://www.chemistry.manchester.ac.uk/epr/about/
Entry Requirements
Applicants must have obtained or be about to obtain a First or Upper Second class UK honours degree, or the equivalent qualifications gained outside the UK, in an appropriate area of science, engineering or technology.
Applicants interested in this project should make direct contact with the Primary Supervisor to arrange to discuss the project further as soon as possible.
How To Apply
To be considered for this project you MUST submit a formal online application form - full details on how to apply can be found on the BBSRC DTP website www.manchester.ac.uk/bbsrcdtpstudentships
Equality, Diversity and Inclusion
Equality, diversity and inclusion is fundamental to the success of The University of Manchester, and is at the heart of all of our activities. The full Equality, diversity and inclusion statement can be found on the website https://www.bmh.manchester.ac.uk/study/research/apply/equality-diversity-inclusion/
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