Dr Stéphane Mesnage

School of Biosciences

Senior Lecturer

Dr Stephane Mesnage
Profile picture of Dr Stephane Mesnage
s.mesnage@sheffield.ac.uk
+44 114 222 4405

Full contact details

Dr Stéphane Mesnage
School of Biosciences
Firth Court
Western Bank
Sheffield
S10 2TN
Profile
  • 2019 - present: Senior Lecturer, School of Biosciences, University of Sheffield, UK
  • 2012 - 2018: Lecturer, Dept. of Molecular Biology and Biotechnology, University of Sheffield, UK
  • 2010 - 2012: Marie Curie (IEF) Fellow, Dept. of Molecular Biology and Biotechnology, University of Sheffield, UK
  • 2002 - 2010: INSERM Chargé de Recherches, University of Paris 6, France
  • 2000 - 2002: EMBO Research assistant, John Innes Centre, Norwich, UK
  • 1996 - 2000: PhD, Pasteur Institute, Paris, France
Research interests

My work is focused on the study of the bacterial cell wall using essentially two Gram-positive pathogens (Staphylococcus aureusEnterococcus faecalis) as models. I am interested in three major areas of research.

Analysis of protein-cell wall interactions

The essential cell wall peptidoglycan polymer network forms a scaffold for the display of a large variety of proteins involved in cell division, metabolism, cell-cell communication and in pathogen interaction with the host. Although several multimodular cell wall binding domains have been identified to date in bacteria, both their structural organisation and binding mechanism (binding motif, subcellular localisation) have remained elusive.

The aim of this project is to understand the molecular mechanisms underpinning protein-bacterial cell wall interactions and subcellular localisation of surface proteins.

Beyond these fundamental questions, the aim is to develop a translational research program to (i) develop peptidoglycan binding domains as new functional probes to provide a nanoscale topological map of the cell surface; (ii) provide an experimental rationale to engineer and target therapeutic molecules against pathogens.

This project involves multidisciplinary approaches across several disciplines including molecular biology, biochemistry, biophysics, structural biology and state-of-the-art super resolution microscopy. 

Control of peptidoglycan hydrolysis during growth

During growth, the insertion of new precursors and separation of daughter cells requires limited cleavage of the peptidoglycan molecule by peptidoglycan hydrolases.

Depending on the bond they cleave, these enzymes are classified as N-acetylmuramidases, N-acetylglucosaminidases, N-acetylmuramoyl L-alanine amidases, endopeptidases or carboxypeptidases (Fig. 2). In addition to their contribution to cell growth and division, some peptidoglycan hydrolases play a role in adhesion and in amplification of the inflammatory response by releasing muramyl-peptides.

The lytic activity of some peptidoglycan hydrolases (called autolysins) is potentially lethal and can lead to cell lysis and death. How bacteria control autolytic activities is poorly understood.

I am studying several mechanisms that may be responsible for the control of autolytic activities: (i) the subcellular targeting of autolysins; (ii) the impact of peptidoglycan structural properties on autolytic activities and (iii) the contribution of the electrochemical gradient across the cytoplasmic membrane to the control of peptidoglycan hydrolysis.

We routinely analyse peptidoglycan hydrolase substrate specificity by a combination of rp-HPLC and mass spectrometry. The enzymatic activity of autolysins is measured spectrophotometrically.

Role of Enterococcus faecalis cell wall metabolism in the interaction with the host

Lysozyme is the major component of the innate immune system and represents the first line of defense against pathogens. E. faecalis is highly resistant to lysozyme, with a minimal inhibitory concentration (MIC) above 50 mg/ml.

This extreme resistance has been partly explained by an additive effect of peptidoglycan O-acetylation and de-N-acetylation, as well as teichoic acid D-alanylation. It was also shown that full lysozyme resistance requires a signalling cascade involving the extracytoplasm

ic function (ECF) sigma factor SigV. Interestingly, E. faecalis strains harbouring multiple deletions in these pathways are still resistant to lysozyme. The identification of novel factors involved in lysozyme resistance is currently underway using random transposon mutagenesis. The mutants isolated will be further characterised in vivo.

Publications

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Journal articles

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Journal articles

Chapters

Conference proceedings papers

  • Bern MW, Beniston R & Mesnage S (2016) Automated Analysis of Bacterial Peptidoglycan Structure. GLYCOBIOLOGY, Vol. 26(12) (pp 1457-1457) RIS download Bibtex download

Preprints