Peptidoglycan (PGN) is the major component of the bacterial cell wall, a structure that is essential for the physical integrity and shape of the cell. Bacteria maintain cell shape by directing PGN incorporation to distinct regions of the cell, namely, through the localization of late-stage PGN synthesis proteins. These include two key protein families, SEDS transglycosylases and bPBP transpeptidases, proposed to function in cognate pairs. Rod-shaped bacteria have two SEDS–bPBP pairs, involved in elongation and division. Here, we elucidate why coccoid bacteria, such as Staphylococcus aureus, also possess two SEDS–bPBP pairs. We determined that S. aureus RodA–PBP3 and FtsW–PBP1 probably constitute cognate pairs of interacting proteins. A lack of RodA–PBP3 resulted in more spherical cells due to deficient sidewall PGN synthesis, whereas depletion of FtsW–PBP1 arrested normal septal PGN incorporation. Although PBP1 is an essential protein, a mutant lacking PBP1 transpeptidase activity is viable, showing that this protein has a second function. We propose that the FtsW–PBP1 pair has a role in stabilizing the divisome at midcell. In the absence of these proteins, the divisome appears as multiple rings or arcs that drive lateral PGN incorporation, leading to cell elongation. We conclude that RodA–PBP3 and FtsW–PBP1 mediate sidewall and septal PGN incorporation, respectively, and that their activity must be balanced to maintain coccoid morphology.
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The data that support the findings of this study are available from the corresponding author on request.
The in-house developed image analysis software is available in the GitHub repository: https://github.com/BacterialCellBiologyLab/PyFRET.
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We thank L. Moreira for hosting laboratory work at the Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal, S. R. Filipe (FCT-NOVA) and R. Carballido-Lopez (INRA) for helpful discussions, S. Bonucci and E. M. Tranfield (Electron Microscopy Facility, IGC) for technical expertise and sample processing, and A. Bernardo, I. Jorge, D. Kądziołka and K. Witana for help in the construction of some plasmids. This study was funded by the European Research Council through grant ERC-2017-CoG-771709 (to M.G.P.), Project LISBOA-01-0145FEDER-007660 Microbiologia Molecular, Estrutural e Celular (to ITQB-NOVA), the Portuguese Platform of Bioimaging PPBI-POCI-01-0145-FEDER-022122, researcher contract no. IF/00386/2015 (to F.F.) and FCT fellowships SFRH/BPD/95031/2013 (to N.T.R.) and SFRH/BD/52204/2013 (to A.C.T.). Whole-genome sequencing analysis at the Genomics Unit of Instituto Gulbenkian de Ciencia was supported by the ONEIDA project (LISBOA-01-0145-FEDER-016417) co-funded by FEEI—‘Fundos Europeus Estruturais e de Investimento’ from ‘Programa Operacional Regional Lisboa 2020’—and by national funds from FCT—‘Fundação para a Ciência e a Tecnologia’.
The authors declare no competing interests.
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Supplementary Figures 1–11, Supplementary Tables 1–3, Supplementary Video legends and Supplementary References.
ColFtsWi time-lapse imaging.
ColPBP1i time-lapse imaging.
ColPBP1TP time-lapse imaging.
ColPBP1TP time-lapse imaging.
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Reichmann, N.T., Tavares, A.C., Saraiva, B.M. et al. SEDS–bPBP pairs direct lateral and septal peptidoglycan synthesis in Staphylococcus aureus. Nat Microbiol 4, 1368–1377 (2019). https://doi.org/10.1038/s41564-019-0437-2
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