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Overlapping and essential roles for molecular and mechanical mechanisms in mycobacterial cell division

Nature Physics volume 16, pages 57–62 (2020)Cite this article

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Abstract

Mechanisms to control cell division are essential for cell proliferation and survival1. Bacterial cell growth and division require the coordinated activity of peptidoglycan synthases and hydrolytic enzymes2,3,4 to maintain mechanical integrity of the cell wall5. Recent studies suggest that cell separation is governed by mechanical forces6,7. How mechanical forces interact with molecular mechanisms to control bacterial cell division in space and time is poorly understood. Here, we use a combination of atomic force microscope imaging, nanomechanical mapping and nanomanipulation to show that enzymatic activity and mechanical forces serve overlapping and essential roles in mycobacterial cell division. We find that mechanical stress gradually accumulates in the cell wall, concentrated at the future division site, culminating in rapid (millisecond) cleavage of nascent sibling cells. Inhibiting cell wall hydrolysis delays cleavage; conversely, locally increasing cell wall stress causes instantaneous and premature cleavage. Cells deficient in peptidoglycan hydrolytic activity fail to locally decrease their cell wall strength and undergo natural cleavage, instead forming chains of non-growing cells. Cleavage of these cells can be mechanically induced by local application of stress with an atomic force microscope. These findings establish a direct link between actively controlled molecular mechanisms and passively controlled mechanical forces in bacterial cell division.

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Fig. 1: The PCF is a nanoscale cell-surface feature that marks the future division site in M. smegmatis.
Fig. 2: Daughter cell cleavage occurs with millisecond kinetics by rupture around the PCF.
Fig. 3: Turgor pressure drives cell cleavage through stress concentration at the PCF.
Fig. 4: AFM-applied local stress on the PCF can substitute for an essential cell division enzyme (RipA).

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Data availability

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

References

  1. Cabeen, M. T. & Jacobs-Wagner, C. Bacterial cell shape. Nat. Rev. Microbiol. 3, 601–610 (2005).

    Article  Google Scholar 

  2. Egan, A. J. & Vollmer, W. The physiology of bacterial cell division. Ann. NY Acad. Sci. 1277, 8–28 (2013).

    Article  ADS  Google Scholar 

  3. Gray, A. N. et al. Coordination of peptidoglycan synthesis and outer membrane constriction during Escherichia coli cell division. eLife 4, e07118 (2015).

    Article  Google Scholar 

  4. Van Der Hofstadt, M., Hüttener, M., Juárez, A. & Gomila, G. Nanoscale imaging of the growth and division of bacterial cells on planar substrates with the atomic force microscope. Ultramicroscopy 154, 29–36 (2015).

    Article  Google Scholar 

  5. Chao, M. C. et al. Protein complexes and proteolytic activation of the cell wall hydrolase RipA regulate septal resolution in mycobacteria. PLoS Pathog. 9, e1003197 (2013).

    Article  Google Scholar 

  6. Zhou, X. et al. Mechanical crack propagation drives millisecond daughter cell separation in Staphylococcus aureus. Science 348, 574–578 (2015).

    Article  ADS  Google Scholar 

  7. Zhou, X., Halladin, D. K. & Theriot, J. A. Fast mechanically driven daughter cell separation is widespread in Actinobacteria. mBio 7, e00952-16 (2016).

    Article  Google Scholar 

  8. Monteiro, J. M. et al. Cell shape dynamics during the staphylococcal cell cycle. Nat. Commun. 6, 8055 (2015).

    Article  ADS  Google Scholar 

  9. Vijay, S., Anand, D. & Ajitkumar, P. Unveiling unusual features of formation of septal partition and constriction in mycobacteria–an ultrastructural study. J. Bacteriol. 194, 702–707 (2012).

    Article  Google Scholar 

  10. Takade, A., Takeya, K., Taniguchi, H. & Mizuguchi, Y. Electron microscopic observations of cell division in Mycobacterium vaccae V1. J. Gen. Microbiol. 129, 2315–2320 (1983).

    Google Scholar 

  11. Dahl, J. L. Electron microscopy analysis of Mycobacterium tuberculosis cell division. FEMS Microbiol. Lett. 240, 15–20 (2004).

    Article  Google Scholar 

  12. Koch, A. L. Biophysics of bacterial walls viewed as stress-bearing fabric. Microbiol. Rev. 52, 337–353 (1988).

    Google Scholar 

  13. Dufrêne, Y. F. Atomic force microscopy in microbiology: new structural and functional insights into the microbial cell surface. mBio 5, e01363-14 (2014).

    Article  Google Scholar 

  14. Herman-Bausier, P. et al. Mechanical strength and inhibition of the Staphylococcus aureus collagen-binding protein Cna. mBio 7, e01529-16 (2016).

    Article  Google Scholar 

  15. Oh, Y. J. et al. Curli mediate bacterial adhesion to fibronectin via tensile multiple bonds. Sci. Rep. 6, 33909 (2016).

    Article  ADS  Google Scholar 

  16. Deng, Y., Sun, M. & Shaevitz, J. W. Direct measurement of cell wall stress stiffening and turgor pressure in live bacterial cells. Phys. Rev. Lett. 107, 158101 (2011).

    Article  ADS  Google Scholar 

  17. Touhami, A., Jericho, M. H. & Beveridge, T. J. Atomic force microscopy of cell growth and division in Staphylococcus aureus. J. Bacteriol. 186, 3286–3295 (2004).

    Article  Google Scholar 

  18. Dover, R. S., Bitler, A., Shimoni, E., Trieu-Cuot, P. & Shai, Y. Multiparametric AFM reveals turgor-responsive net-like peptidoglycan architecture in live streptococci. Nat. Commun. 6, 7193 (2015).

    Article  ADS  Google Scholar 

  19. Eskandarian, H. A. et al. Division site selection linked to inherited cell surface wave troughs in mycobacteria. Nat. Microbiol. 2, 17094 (2017).

    Article  Google Scholar 

  20. Turner, R. D., Hurd, A. F., Cadby, A., Hobbs, J. K. & Foster, S. J. Cell wall elongation mode in Gram-negative bacteria is determined by peptidoglycan architecture. Nat. Commun. 4, 1496 (2013).

    Article  ADS  Google Scholar 

  21. Bailey, R. G. et al. The interplay between cell wall mechanical properties and the cell cycle in Staphylococcus aureus. Biophys. J. 107, 2538–2545 (2014).

    Article  ADS  Google Scholar 

  22. Loskill, P. et al. Reduction of the peptidoglycan crosslinking causes a decrease in stiffness of the Staphylococcus aureus cell envelope. Biophys. J. 107, 1082–1089 (2014).

    Article  ADS  Google Scholar 

  23. Santi, I., Dhar, N., Bousbaine, D., Wakamoto, Y. & McKinney, J. D. Single-cell dynamics of the chromosome replication and cell division cycles in mycobacteria. Nat. Commun. 4, 2470 (2013).

    Article  ADS  Google Scholar 

  24. Dufrêne, Y. F., Martínez-Martín, D., Medalsy, I., Alsteens, D. & Müller, D. J. Multiparametric imaging of biological systems by force–distance curve-based AFM. Nat. Methods 10, 847–854 (2013).

    Article  Google Scholar 

  25. Sun, S. X. & Jiang, H. Physics of bacterial morphogenesis. Microbiol. Mol. Biol. Rev. 75, 543–565 (2011).

    Article  Google Scholar 

  26. Francius, G., Domenech, O., Mingeot-Leclercq, M. P. & Dufrêne, Y. F. Direct observation of Staphylococcus aureus cell wall digestion by lysostaphin. J. Bacteriol. 190, 7904–7909 (2008).

    Article  Google Scholar 

  27. Wheeler, R. et al. Bacterial cell enlargement requires control of cell wall stiffness mediated by peptidoglycan hydrolases. mBio 6, e00660-15 (2015).

    Article  Google Scholar 

  28. Ruggiero, A. et al. Structure and functional regulation of RipA, a mycobacterial enzyme essential for daughter cell separation. Structure 18, 1184–1190 (2010).

    Article  Google Scholar 

  29. Hett, E. C. et al. A partner for the resuscitation-promoting factors of Mycobacterium tuberculosis. Mol. Microbiol. 66, 658–668 (2007).

    Article  Google Scholar 

  30. Hett, E. C., Chao, M. C., Deng, L. L. & Rubin, E. J. A mycobacterial enzyme essential for cell division synergizes with resuscitation-promoting factor. PLoS Pathog. 4, e1000001 (2008).

    Article  Google Scholar 

  31. Jiang, Y., Liang, X., Guo, M., Cao, Y. & Cai, S. Fracture mechanics modeling of popping event during daughter cell separation. Biomech. Model. Mechanobiol. 17, 1131–1137 (2018).

    Article  Google Scholar 

  32. Thanky, N. R., Young, D. B. & Robertson, B. D. Unusual features of the cell cycle in mycobacteria: polar-restricted growth and the snapping-model of cell division. Tuberculosis 87, 231–236 (2007).

    Article  Google Scholar 

  33. Smolyakov, G., Formosa-Dague, C., Severac, C., Duval, R. E. & Dague, E. High speed indentation measures by FV, QI and QNM introduce a new understanding of bionanomechanical experiments. Micron 85, 8–14 (2016).

    Article  Google Scholar 

  34. Atilgan, E., Magidson, V., Khodjakov, A. & Chang, F. Morphogenesis of the fission yeast cell through cell wall expansion. Curr. Biol. 25, 2150–2157 (2015).

    Article  Google Scholar 

  35. Meniche, X. et al. Subpolar addition of new cell wall is directed by DivIVA in mycobacteria. Proc. Natl Acad. Sci. USA 111, E3243–E3251 (2014).

    Article  Google Scholar 

  36. Odermatt, P. D. et al. High-resolution correlative microscopy: bridging the gap between single molecule localization microscopy and atomic force microscopy. Nano Lett. 15, 4896–4904 (2015).

    Article  ADS  Google Scholar 

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Acknowledgements

We thank E. J. Rubin for generously providing the RipA conditional depletion strain of M. smegmatis. This research was supported in part by grants to G.E.F. from the Swiss National Science Foundation (205321_134786 and 205320_152675), from the European Union FP7/2007-2013/ERC under grant agreement no. 307338-NaMic, H2020 - UE Framework Programme for Research & Innovation (2014-2020); ERC-2017-CoG; InCell; Project number 773091, from the Commission for Technology and Innovation under CTI no. 18330.1 PFNM-NM, and by a grant to J.D.M. from the Swiss National Science Foundation (310030B_176397). H.A.E. was supported by an EMBO Long Term Fellowship (191-2014) and an EMBO Advanced Long Term Fellowship (750-2016).

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Authors and Affiliations

  1. Laboratory for Bio- and Nano-Instrumentation, School of Engineering, Swiss Federal Institute of Technology Lausanne, Lausanne, Switzerland

    Pascal D. Odermatt, Mélanie T. M. Hannebelle, Haig A. Eskandarian, Adrian P. Nievergelt & Georg E. Fantner

  2. Laboratory of Microbiology and Microtechnology, School of Life Sciences, Swiss Federal Institute of Technology Lausanne, Lausanne, Switzerland

    Mélanie T. M. Hannebelle, Haig A. Eskandarian & John D. McKinney

Authors
  1. Pascal D. Odermatt
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Contributions

P.D.O., G.E.F. and J.D.M. conceptualized the study. P.D.O., M.T.M.H. and H.A.E. developed imaging protocols. P.D.O., M.T.M.H., G.E.F. and J.D.M. designed experiments. P.D.O. and M.T.M.H. performed the experiments and analysed the data. A.P.N. participated in building the instrument and simulations. P.D.O., M.T.M.H., G.E.F. and J.D.M. wrote the manuscript.

Corresponding authors

Correspondence to John D. McKinney or Georg E. Fantner.

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The authors declare no competing interests.

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Peer review information Nature Physics thanks Yves Dufrene and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–17, Table 1 and refs. 1 and 2.

Reporting Summary

Supplementary Video 1

A time sequence of M. smegmatis cells undergoing abrupt cleavage imaged in QNM with a low force setpoint. Frame rate: 5 min.

Supplementary Video 2

A time sequence of stiffness increase at the PCF imaged in QNM with a higher force setpoint. Frame rate: 5 min.

Supplementary Video 3

A time sequence of an alternative mechanism of separation of sibling cells when one sibling cell is deflated by piercing with the AFM cantilever tip. Sibling cells do not undergo rapid cell cleavage; instead, the surviving sibling gradually sheds the deflated sibling.

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Odermatt, P.D., Hannebelle, M.T.M., Eskandarian, H.A. et al. Overlapping and essential roles for molecular and mechanical mechanisms in mycobacterial cell division. Nat. Phys. 16, 57–62 (2020). https://doi.org/10.1038/s41567-019-0679-1

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