Details
This project focuses on the structure/function analysis of a surface polysaccharide produced by enterococci, required for normal cell growth, division, resistance to antibiotics and pathogenesis. This enterococcal polysaccharide antigen (EPA) is encoded by two gene clusters: (i) 18 genes extremely conserved encoding a core synthetic machinery and (ii) a set of genes variable between strains, responsible for the decoration of the polysaccharide backbone. We have built a mutant with a complete deletion of the epa variable region and shown that the decoration of EPA (but not its core structure) is essential for the biological activity of this polymer. We propose to investigate EPA structural properties and dissect the contribution EPA decoration to bacterial physiology and antibiotic resistance and recognition by bacteriophages.
We will take a multidisciplinary approach to explore EPA structure and to understand how EPA controls the distribution and activity of surface proteins involved in cell wall assembly and resistance to antibiotics. The project will involve state-of-the-art methodologies including super-resolution microscopy (fluorescence and electron microscopy), structural glycobiology (NMR, mass spectrometry) and conventional approaches to study bacterial physiology and antimicrobial resistance. The candidate will work with members of the supervisors’ laboratories studying bacterial cell surface assembly, host-pathogen interaction, and bacteriophages.
About the DTP
This studentship is offered as part of the White Rose BBSRC Doctoral Training Partnership (DTP) in Mechanistic Biology, which brings together the research of the world-class molecular and cellular bioscience centres at the White Rose universities of Leeds, Sheffield and York.
Our mission is to train excellent bio-scientists who understand how living systems work
and can innovate to address global challenges, such as the impact of climate change, a healthier old age, sustainable food production, land use and energy production.
What is on offer?
This is a core studentship for entry in October 2024.
Join us and you will receive a 4-year, funded PhD programme of research and skills training, with cross-disciplinary supervision, plus a structured programme of cohort-wide training and networking events. A highlight is the annual symposium, which is planned and delivered by students.
A unique part of your training will be the Professional Internships for PhD Students (PIPS), where you will spend three months at a host organisation of your choosing, gaining experience of work in a professional environment, and acquiring transferable skills that will be beneficial in your future career.
How to apply – Expression of Interest
Students may apply for up to three projects anywhere in the Doctoral Training Partnership (DTP). Applications will be to the DTP centrally, using an online Expression of Interest (EoI). The EoI will include:
§ CV information; not submitted separately
§ Equality, Diversity and Inclusion (EDI) data
§ Names of two referees
Deadline for EoIs is midnight Sunday 7th January 2024.
Submit EoIs using this link: https://leeds.onlinesurveys.ac.uk/white-rose-bbsrc-dtp-expression-of-interest-form
Shortlisted candidates will be required to make formal applications to the Graduate School at each institution, supplying the necessary paperwork.
Interviews will be held either Friday 2nd and Monday 5th to Friday 9th February, or Monday 19th to Friday 23rd and Monday 26th February 2024, in-person at Leeds, Sheffield and York, with a panel representing all 3 Universities. Shortlisted candidates will be notified of a specific time/date to attend. If you have applied for more than one project and are selected for interview, you will be interviewed only once.
Funding Notes
Appointed candidates will be fully funded for 4 years:
Tax-free annual stipend at the UKRI rate. The rate for starters in 2023/24 was £18,622. (Rates for 2024/25 starters are not yet available).
UKRI tuition fees – These are paid directly to the host institution.
A Research Training and Support Grant
An allowance for Fieldwork/Conference/Travel
A Professional Internship for PhD Students (PIPS) allowance
Not all projects will be funded; the DTP will appoint a limited number of candidates via a competitive process.
References
Davis JL, Hounslow AM, Baxter NJ, Mesnage S, Williamson MP.
1H, 13C, and 15N resonance assignments of a conserved putative cell wall binding domain from Enterococcus faecalis.
Biomol NMR Assign. 2022, 16(2):247-251. doi: 10.1007/s12104-022-10087-2.
Gonzalez-Delgado LS, Waters-Morgan H, Salamaga B, Robertson AJ, Hounslow AM, Jagielska E, Sabala I, Williamson MP, Lovering AL, Mesnage S. Two site recognition of Staphylococcus aureus peptidoglycan by lysostaphin SH3b.
Nature Chem Biol. 2020, 16’(1): 24-30. doi: 10.1038/s41589-019-0393-4.
Smith RE, Salamaga B, Szkuta P, Hajdamowicz N, Prajsnar TK, Bulmer GS, Fontaine T, Kołodziejczyk J, Herry JM, HounslowM, Williamson MP, Serror P, Mesnage S
Decoration of the enterococcal polysaccharide antigen EPA is essential for virulence, cell surface charge and interaction with effectors of the innate immune system.
PLoS Pathog. 2019, 15(5): e1007730
Al-Zubidi M, Widziolek M, Court EK, Gains AF, Smith RE, Ansbro K, Alrafaie A, Evans C, Murdoch C, Mesnage S, Douglas CWI, Rawlinson A, Stafford GP. Identification of novel bacteriophages with therapeutic potential that target Enterococcus faecalis
Infection and immunity. 2019, 87 (11), e00512-19.
Davis JL, Hounslow AM, Baxter NJ, Mesnage S, Williamson MP.
1H, 13C, and 15N resonance assignments of a conserved putative cell wall binding domain from Enterococcus faecalis.
Biomol NMR Assign. 2022 16(2):247-251. doi: 10.1007/s12104-022-10087-2.
Hobbs B, Drant J, Williamson MP.
The measurement of binding affinities by NMR chemical shift perturbation.
J Biomol NMR. 2022 76(4):153-163. doi: 10.1007/s10858-022-00402-3.
Fowler NJ, Albalwi MF, Lee S, Hounslow AM, Williamson MP.
Improved methodology for protein NMR structure calculation using hydrogen bond restraints and ANSURR validation: The SH2 domain of SH2B1.
Structure. 2023 31(8):975-986.e3. doi: 10.1016/j.str.2023.05.012.
Southam HM, Williamson MP, Chapman JA, Lyon RL, Trevitt CR, Henderson PJF, Poole RK.
'Carbon-Monoxide-Releasing Molecule-2 (CORM-2)' Is a Misnomer: Ruthenium Toxicity, Not CO Release, Accounts for Its Antimicrobial Effects.
Antioxidants (Basel). 2021 10(6):915. doi: 10.3390/antiox10060915.
Satur MJ, Urbanowicz PA, Spencer DIR, Rafferty J, Stafford GP
Structural and functional characterisation of a stable, broad-specificity multimeric sialidase from the oral pathogen Tannerella forsythia.
Biochem J. 2022 479(17):1785-1806. doi: 10.1042/BCJ20220244.
Court EK, Chaudhuri RR, Kapoore RV, Villa RX, Pandhal J, Biggs CA, Stafford GP
Looking through the FOG: microbiome characterization and lipolytic bacteria isolation from a fatberg site.
Microbiology. 2021 167(12):001117. doi: 10.1099/mic.0.001117
Al-Zubidi M, Widziolek M, Court EK, Gains AF, Smith RE, Ansbro K, Alrafaie A, Evans C, Murdoch C, Mesnage S, Douglas CWI, Rawlinson A, Stafford GP
Identification of Novel Bacteriophages with Therapeutic Potential That Target Enterococcus faecalis
Infection and immunity. 2019, 87 (11), e00512-19.
Green CA, Kamble NS, Court EK, Bryant OJ, Hicks MG, Lennon C, Fraser GM, Wright PC, Stafford GP
Engineering the flagellar type III secretion system: improving capacity for secretion of recombinant protein.
Microb Cell Fact. 2019 18(1):10. doi:10.1186/s12934-019-1058-4.