Wall proficient e. Coli capable of sustained growth in the absence of the z-ring division machine

ABSTRACT The peptidoglycan cell wall is a major protective external sheath in bacteria and a key target for antibiotics1. Peptidoglycan is present in virtually all bacteria, suggesting that

it was probably present in the last bacterial common ancestor2. Cell wall expansion is orchestrated by cytoskeletal proteins related to actin (MreB) and tubulin (FtsZ)3. FtsZ is a key

essential player in a highly organized division machine that directs an invaginating annulus of cell wall peptidoglycan. The recent discovery that cell-wall-less bacteria (L-forms) can grow

and divide independently of FtsZ4,5, provided a means of generating an _ftsZ_ null mutant of _Escherichia coli_. Remarkably, we have been able to isolate variants of _E. coli_ that lack FtsZ

but are capable of efficient growth in a walled state. Genetic analysis reveals that a combination of mutations is needed for this phenotype. Importantly, the suppressive mutations lead to

a major cell shape change, from the normal cylindrical shape to a branched and bulging, ramified shape, which we call ‘coli-flower’. The results highlight the versatility of bacterial cells

and illustrate possible evolutionary routes leading to the emergence of specialized bacteria, such as pathogenic _Chlamydia_ or aquatic Planctomycetes, that lack FtsZ but retain the cell

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Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS SEPTAL WALL SYNTHESIS IS SUFFICIENT TO CHANGE AMEBA-LIKE CELLS INTO UNIFORM OVAL-SHAPED CELLS IN _ESCHERICHIA COLI_ L-FORMS

Article Open access 26 November 2024 TEICHOIC ACIDS ANCHOR DISTINCT CELL WALL LAMELLAE IN AN APICALLY GROWING BACTERIUM Article Open access 17 June 2020 DETERMINING THE RATE-LIMITING

PROCESSES FOR CELL DIVISION IN _ESCHERICHIA COLI_ Article Open access 16 November 2024 REFERENCES * Schneider, T. & Sahl, H. G. Lipid II and other bactoprenol-bound cell wall precursors

as drug targets. _Curr. Opin. Invest. Drugs_ 11, 157–164 (2010). CAS  Google Scholar  * Errington, J. L-form bacteria, cell walls and the origins of life. _Open Biol_. 3, 120143 (2013).

Article  Google Scholar  * Typas, A., Banzhaf, M., Gross, C. A. & Vollmer, W. From the regulation of peptidoglycan synthesis to bacterial growth and morphology. _Nature Rev. Microbiol_.

10, 123–136 (2012). Article  CAS  Google Scholar  * Leaver, M., Dominguez-Cuevas, P., Coxhead, J. M., Daniel, R. A. & Errington, J. Life without a wall or division machine in _Bacillus

subtilis_. _Nature_ 457, 849–853 (2009). Article  CAS  Google Scholar  * Mercier, R., Kawai, Y. & Errington, J. General principles for the formation and proliferation of a wall-free

(L-form) state in bacteria. _eLife_ 3, 04629 (2014). Article  Google Scholar  * Pilhofer, M. _et al._ Discovery of chlamydial peptidoglycan reveals bacteria with murein sacculi but without

FtsZ. _Nature Commun_. 4, 2856 (2013). Article  Google Scholar  * Liechti, G. W. _et al._ A new metabolic cell-wall labelling method reveals peptidoglycan in _Chlamydia trachomatis_.

_Nature_ 506, 507–510 (2014). Article  CAS  Google Scholar  * Jeske, O. _et al._ Planctomycetes do possess a peptidoglycan cell wall. _Nature Commun_. 6, 7116 (2015). Article  CAS  Google

Scholar  * Kawai, Y., Mercier, R. & Errington, J. Bacterial cell morphogenesis does not require a preexisting template structure. _Curr. Biol_. 24, 863–867 (2014). Article  CAS  Google

Scholar  * Ranjit, D. K. & Young, K. D. The Rcs stress response and accessory envelope proteins are required for _de novo_ generation of cell shape in _Escherichia coli_. _J. Bacteriol_.

195, 2452–2462 (2013). Article  CAS  Google Scholar  * Billings, G. _et al._ _De novo_ morphogenesis in L-forms via geometric control of cell growth. _Mol. Microbiol_. 93, 883–896 (2014).

Article  CAS  Google Scholar  * Dai, K. & Lutkenhaus, J. _ftsZ_ is an essential cell division gene in _Escherichia coli_. _J. Bacteriol._ 173, 3500–3506 (1991). Article  CAS  Google

Scholar  * Paradis-Bleau, C. _et al._ Lipoprotein cofactors located in the outer membrane activate bacterial cell wall polymerases. _Cell_ 143, 1110–1120 (2010). Article  CAS  Google Scholar

  * Typas, A. _et al._ Regulation of peptidoglycan synthesis by outer-membrane proteins. _Cell_ 143, 1097–1109 (2010). Article  CAS  Google Scholar  * Gray, A. N. _et al._ Coordination of

peptidoglycan synthesis and outer membrane constriction during _Escherichia coli_ cell division. _eLife_ 4, 07118 (2015). Article  Google Scholar  * Braun, V. & Bosch, V. Repetitive

sequences in the murein-lipoprotein of the cell wall of _Escherichia coli_. _Proc. Natl Acad. Sci. USA_ 69, 970–974 (1972). Article  CAS  Google Scholar  * Sambrook, J., Fritsch, E. F. &

Maniatis, T. _Molecular Cloning: A Laboratory Manual_ (Cold Spring Harbor Laboratory Press, 1989). Google Scholar  * Datsenko, K. A. & Wanner, B. L. One-step inactivation of chromosomal

genes in _Escherichia coli_ K-12 using PCR products. _Proc. Natl Acad. Sci. USA_ 97, 6640–6645 (2000). Article  CAS  Google Scholar  * Jensen, R. B., Grohmann, E., Schwab, H., Diaz-Orejas,

R. & Gerdes, K. Comparison of ccd of F, parDE of RP4, and parD of R1 using a novel conditional replication control system of plasmid R1. _Mol. Microbiol_. 17, 211–220 (1995). Article 

CAS  Google Scholar  * Shen, Z. _et al._ MPprimer: a program for reliable multiplex PCR primer design. _BMC Bioinformatics_ 11, 143 (2010). Article  Google Scholar  Download references

ACKNOWLEDGEMENTS The authors thank T. Mignot for hosting parts of the work in his laboratory, W. Vollmer for the gift of the plasmid pBAD33-_lpoB_ and comments on the manuscript, H. Strahl

and L. Espinosa for discussions and D.W. Adams for critical reading of the manuscript. This work was funded by European Research Council Advanced Investigator Grants (nos 250363 and

BH141574) to J.E. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Medical School, Newcastle University,

Richardson Road, Newcastle upon Tyne NE2 4AX, UK Romain Mercier, Yoshikazu Kawai & Jeff Errington * Laboratoire de Chimie Bactérienne, Institut de Microbiologie de la Méditerranée,

CNRS-Aix-Marseille University, 31 Chemin Joseph Aiguier, 13009 Marseille, France Romain Mercier Authors * Romain Mercier View author publications You can also search for this author inPubMed

 Google Scholar * Yoshikazu Kawai View author publications You can also search for this author inPubMed Google Scholar * Jeff Errington View author publications You can also search for this

author inPubMed Google Scholar CONTRIBUTIONS R.M. designed and performed the experiments. R.M. and Y.K. analysed the data. R.M. and J.E. wrote the manuscript. CORRESPONDING AUTHORS

Correspondence to Romain Mercier or Jeff Errington. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY

INFORMATION Supplementary Data 1–4, Supplementary Figures 1–8, Supplementary Tables 1–3, Supplementary References (PDF 7045 kb) SUPPLEMENTARY VIDEO 1 Time-lapse series showing ΔftsZsup5

cells growing on NA 20mM Mg2+, from which the panels in Figure 2b were obtained. Phase contrast images were acquired automatically every minute for about 279 min. Scale bar is 3 μm. (AVI

1117 kb) SUPPLEMENTARY VIDEO 2 Time-lapse series showing ΔftsZsup1 cell growing on NA 20mM Mg2+, from which the panels in Figure 2b were obtained. Phase contrast images were acquired

automatically every minute for about 279 min. Scale bar is 3 μm. (AVI 2475 kb) SUPPLEMENTARY VIDEO 3 Time-lapse series showing ΔftsZsup7 cell growing on NA 20mM Mg2+. Phase contrast images

were acquired automatically every minute for about 109 min. Scale bar is 3 μm. (AVI 810 kb) SUPPLEMENTARY VIDEO 4 Time-lapse series showing ΔftsZsup7 cell dividing on NA 20mM Mg2+, from

which the panels in Figure 2b were obtained. Phase contrast images were acquired automatically every minute for about 69 min. Scale bar is 3 μm. (AVI 297 kb) SUPPLEMENTARY VIDEO 5 Time-lapse

series showing ΔftsZsup7 cell dividing on NA + 20mM Mg2+, from which the panels in Figure 2c were obtained. Phase contrast images were acquired automatically every minute for about 90 min.

Scale bar is 3 μm. (AVI 419 kb) SUPPLEMENTARY VIDEO 6 Time-lapse series showing RM445 (ΔftsZ/Δlpp/ΔlpoBsup) cell, bearing the plasmid pBAD33-lpoB cell growing on NA + 20mM Mg2+ref with

glucose (lpp expression repressed). Phase contrast images were acquired automatically every minute for about 209 min. Scale bar is 3 μm. (AVI 3149 kb) SUPPLEMENTARY VIDEO 7 Time-lapse series

showing RM445 (ΔftsZ/Δlpp/ΔlpoBsup), bearing the plasmid pBAD33-lpoB cell growing on NA + 20mM Mg2+ with lpoB expression induced (arabinose), from which the panels in Figure 3d were

obtained. Phase contrast images were acquired automatically every minute for about 209 min. Scale bar is 3 μm. (AVI 3477 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS

ARTICLE CITE THIS ARTICLE Mercier, R., Kawai, Y. & Errington, J. Wall proficient _E. coli_ capable of sustained growth in the absence of the Z-ring division machine. _Nat Microbiol_ 1,

16091 (2016). https://doi.org/10.1038/nmicrobiol.2016.91 Download citation * Received: 26 October 2015 * Accepted: 09 May 2016 * Published: 27 June 2016 * DOI:

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