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SEDS–bPBP pairs direct lateral and septal peptidoglycan synthesis in Staphylococcus aureus


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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Fig. 1: S.aureus has two cognate SEDS–bPBP pairs of interacting proteins: FtsW–PBP1 and RodA–PBP3.
Fig. 2: RodA and PBP3 have a role in cell elongation.
Fig. 3: FtsW–PBP1 has a role in maintaining the near-spherical shape of cocci.
Fig. 4: FtsW–PBP1 depletion leads to multiple divisome structures that incorporate PGN at the lateral wall and elongate cells.
Fig. 5: Model for the role of SEDS–bPBPs in S.aureus morphogenesis.

Data availability

The data that support the findings of this study are available from the corresponding author on request.

Code availability

The in-house developed image analysis software is available in the GitHub repository:


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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’.

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Authors and Affiliations



N.T.R., A.C.T. and M.G.P. designed the research. N.T.R. and A.C.T. performed all of the experiments, with the exception of the FLIM data acquisition and analysis, which was performed by F.F., and the HPLC muropeptide analysis performed by A.J. B.M.S. developed software for image analysis of the seFRET data. N.T.R., A.C.T. and P.R. constructed the strains. A.J. and P.R. analysed the whole-genome sequencing data. J.M.M. and A.R.P. performed the preliminary experiments and analysed the data. R.G.S. performed the preliminary experiments. M.S.V.N. contributed new reagents (FDAAs). N.T.R., A.C.T. and M.G.P. analysed the overall data. B.M.S. analysed the seFRET data quantified by in-house developed software. N.T.R., A.C.T. and M.G.P. wrote the manuscript.

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Correspondence to Mariana G. Pinho.

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Supplementary information

Supplementary Information

Supplementary Figures 1–11, Supplementary Tables 1–3, Supplementary Video legends and Supplementary References.

Reporting Summary

Supplementary Video 1

ColFtsWi time-lapse imaging.

Supplementary Video 2

ColPBP1i time-lapse imaging.

Supplementary Video 3

ColPBP1TP time-lapse imaging.

Supplementary Video 4

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).

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