The bacterial cell wall is a highly conserved essential component of most bacterial groups. It is the target for our most frequently used antibiotics and provides important small molecules that trigger powerful innate immune responses. The wall is composed of glycan strands crosslinked by short peptides. For many years, the penicillin-binding proteins were thought to be the key enzymes required for wall synthesis. RodA and possibly other proteins in the wider SEDS (shape, elongation, division and sporulation) family have now emerged as a previously unknown class of essential glycosyltranferase enzymes, which play key morphogenetic roles in bacterial cell wall synthesis. We provide evidence in support of this role and the discovery of small natural product molecules that probably target these enzymes. The SEDS proteins have exceptional potential as targets for new antibacterial therapeutic agents.
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This work was funded by grants from the BBSRC (BB/G015902/1, to R.A.D., J.E. and A.G.), ERC (670980, to J.E., Y.K. and K.E.) and the Wellcome Trust (WT098374AIA, to J.E. and L.J.W.) and core resources in Demuris Ltd. The authors thank R. Emmins, who constructed some of the mutants, D. Roberts for help with microfluidic microscopy and R. Sweet for help with the large-scale fermentation run.
J.E. is a director and shareholder at Demuris Ltd. N.A. and J.D. are employees at Demuris Ltd.
Legends for all Supplementary Material and Supplementary Figures 2–5. (PDF 795 kb)
Supplementary Figure 1 and Supplementary Tables 1 and 2. (PDF 845 kb)
Effect of compound 654/A (30 μl) on growth and morphology of the Δ4 strain. Approximately 3 hours after addition of 654/A to Δ4 cells in the microfluidic device, cell growth slows down significantly and extensive cell lysis occurs. (AVI 19450 kb)
Untreated control for growth and morphology of the Δ4 strain. In the absence of compound 654/A, Δ4 cells continue their normal growth up to full confluency. (AVI 39740 kb)
Effect of compound 654/A (30 μl) on growth and morphology of the wild type. Wild-type cells show normal growth at relatively low concentrations of 654/A. (AVI 34112 kb)
Effect of compound 654/A (70 μl) on growth and morphology of the wild type. At relatively high concentrations of 654/A, extensive lysis of wild-type cells occurs, similar to that of the Δ4 cells (see Supplementary Video 1). (AVI 25887 kb)
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Emami, K., Guyet, A., Kawai, Y. et al. RodA as the missing glycosyltransferase in Bacillus subtilis and antibiotic discovery for the peptidoglycan polymerase pathway. Nat Microbiol 2, 16253 (2017). https://doi.org/10.1038/nmicrobiol.2016.253
The transpeptidase PBP2 governs initial localization and activity of the major cell-wall synthesis machinery in E. coli
Structural coordination of polymerization and crosslinking by a SEDS–bPBP peptidoglycan synthase complex
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