Dr A Winter
No more applications being accepted
Funded PhD Project (European/UK Students Only)
About the Project
Pathogenic viruses infect and ultimately kill healthy cells. The aim of identifying, characterizing and targeting interactions of viral proteins with host proteins has recently become urgent as more viral infections become more prevalent in both the developed and developing world. Recent research suggests that several viruses, such as Dengue and Chikungunya, effect entry into host cells via interaction with cell-surface proteins called prohibitins [1-3]. Prohibitins (prohibitin-1 (Phb1) and prohibitin-2 (Phb2)) have previously been shown to be involved in many basic functions within the cell such as cellular signalling, transcriptional control, apoptosis, cellular senescence, cell growth and survival, metabolism, mitochondrial biogenesis and inflammation [4]. Their involvement in infection is a new finding, and the interaction of prohibitins with viral proteins on the cell surface is of major interest.
This studentship is centred on investigating the low resolution solution structure of (a) the prohibitin heterodimer and (b) the complex between the heterodimer and the viral envelope proteins from Dengue virus and Chikungunya virus, amongst others, using structural and biophysical techniques. X-ray crystallography will be mainly used to determine the molecular structures of the respective proteins and their complexes. Small Angle Neutron Scattering (SANS) and Small Angle X-ray Scattering (SAXS) will provide complementing solution structure information on the complexes as well as on the separate subunits involved. This will be complemented by NMR and chemical cross-linking data, providing additional structural restraints that will form the basis for data-driven docking using HADDOCK. The structural models generated will give a holistic view on how prohibitins interact with each other and with viral proteins (see [5, 6]).
Recombinant protein production and preparation will mainly be carried out at the Department for Life Sciences, Keele University. SANS measurements will be carried out at the ILL (Grenoble, France) in collaboration with Dr. Sylvain Prevost, using recombinantly expressed deuterated protein produced in the Deuteration Laboratory of ILL’s Life Sciences group. SAXS and X-ray crystallography will be carried out at ESRF, Grenoble, France and Diamond Light Source, Oxford, UK.
Candidate profile:
Essential Qualifications and Skills - Degree in biological/chemical/natural sciences - minimum 2i classification or equivalent.
An interest in protein production and analysis using biophysical techniques such as X-ray crystallography and Small Angle Neutron Scattering
Self-motivation to undertake advanced research study at PhD level.
Excellent communication, interpersonal and organizational skills.
The ability to work both independently and as part of a team.
Natural inquisitiveness and a flair for problem solving. Willingness to learn new practical skills.
Flexibility to travel to Grenoble, France on a regular basis.
Desirable:
Previous experience in producing protein in recombinant systems.
Evidence of organizational and time management skills.
Funding Notes
100% UK/EU tuition fees for 3 years commencing Academic year 2017/18.
Stipend support for three years at Research Council rates (2017/8 £14,553 per annum). Funding for consumables and conference attendance is available.
Eligibility Criteria:
Applicants must be UK or EU nationals to be eligible for the studentship. BSc in biological/ chemical /natural sciences - minimum degree classification 2i or equivalent. An interest in both laboratory and computational work (protein production and biophysical techniques) is essential - full training will be provided.
References
1. Wintachai, P., et al., Identification of prohibitin as a Chikungunya virus receptor protein. J Med Virol, 2012. 84(11): p. 1757-70.
2. Sharma, A. and A. Qadri, Vi polysaccharide of Salmonella typhi targets the prohibitin family of molecules in intestinal epithelial cells and suppresses early inflammatory responses. Proc Natl Acad Sci U S A, 2004. 101(50): p. 17492-7.
3. Liu, C., et al., Identification of human host proteins contributing to H5N1 influenza virus propagation by membrane proteomics. J Proteome Res, 2012. 11(11): p. 5396-405.
4. Winter, A., O. Kamarainen, and A. Hofmann, Molecular modeling of prohibitin domains. Proteins, 2007. 68(1): p. 353-62.
5. Trewhella, J., Small-angle scattering and 3D structure interpretation. Curr Opin Struct Biol, 2016. 40: p. 1-7.
6. Hennig, J., et al., Combining NMR and small angle X-ray and neutron scattering in the structural analysis of a ternary protein-RNA complex. J Biomol NMR, 2013. 56(1): p. 17-30.