Abstract

In every living organism, cell division requires accurate identification of the division site and placement of the division machinery. In bacteria, this process is traditionally considered to begin with the polymerization of the highly conserved tubulin-like protein FtsZ into a ring that locates precisely at mid-cell1. Over the past decades, several systems have been reported to regulate the spatiotemporal assembly and placement of the FtsZ ring2,3,4,5. However, the human pathogen Streptococcus pneumoniae, in common with many other organisms, is devoid of these canonical systems and the mechanisms of positioning the division machinery remain unknown4,6. Here we characterize a novel factor that locates at the division site before FtsZ and guides septum positioning in pneumococcus. Mid-cell-anchored protein Z (MapZ) forms ring structures at the cell equator and moves apart as the cell elongates, therefore behaving as a permanent beacon of division sites. MapZ then positions the FtsZ ring through direct protein–protein interactions. MapZ-mediated control differs from previously described systems mostly on the basis of negative regulation of FtsZ assembly. Furthermore, MapZ is an endogenous target of the Ser/Thr kinase StkP, which was recently shown to have a central role in cytokinesis and morphogenesis of S. pneumoniae7,8,9. We show that both phosphorylated and non-phosphorylated forms of MapZ are required for proper Z-ring formation and dynamics. Altogether, this work uncovers a new mechanism for bacterial cell division that is regulated by phosphorylation and illustrates that nature has evolved a diversity of cell division mechanisms adapted to the different bacterial clades.

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Acknowledgements

This work was supported by grants from the CNRS, the Université Claude Bernard Lyon 1, the FINOVI foundation, the Agence National de la Recherche (ANR-12-BSV3-0008-01) and the Region Rhône-Alpes (ARC1 and financial support for A.F. (Cluster 10) and C.Z. (CMIRA)). C.L. was supported by a Wellcome Trust Programme Grant to D.S. (WT083469MA), advanced microscopy at Micron Oxford was supported by a Wellcome Trust Strategic Award (091911), and Y.V.B. was supported by a National Institutes of Health grant (GM051986). We acknowledge the contribution of the PLATIM and the Protein Science facilities of the SFR Biosciences Gerland-Lyon Sud (UMS344/US8) and the Centre Technologique des Microstructures de l’Université Lyon 1. We thank C. Chamot for technical assistance in microscopy, S. Uphoff and J. Wiktor for providing help with Matlab and A.-M. Di Guilmi for providing us with the antibody against pneumococcal FtsZ. We thank N. Campo, N. Dubarry and J.-P. Claverys for stimulating discussions and critical reading of the manuscript. N. Campo is also thanked for her help with time-lapse microscopy.

Author information

Author notes

    • Aurore Fleurie
    • , Christian Lesterlin
    •  & Sylvie Manuse

    These authors contributed equally to this work.

    • Chao Zhao

    Present address: Key laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China.

Affiliations

  1. Bases Moléculaires et Structurales des Systèmes Infectieux, IBCP, Université Lyon 1, CNRS, UMR 5086, Lyon 69007, France

    • Aurore Fleurie
    • , Sylvie Manuse
    • , Chao Zhao
    • , Jean-Pierre Lavergne
    • , Christophe Combet
    •  & Christophe Grangeasse
  2. Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK

    • Christian Lesterlin
    •  & David Sherratt
  3. Laboratoire de Biologie Tissulaire et d’Ingénierie Thrérapeutique, IBCP, Université Lyon 1, CNRS, UMR 5305, Lyon 69007, France

    • Caroline Cluzel
  4. Proteome Center Tübingen, University of Tübingen, Auf der Morgenstelle 15, Tübingen 72076, Germany

    • Mirita Franz-Wachtel
    •  & Boris Macek
  5. Departments of Biology and Chemistry, Indiana University, Bloomington, Indiana 47405, USA

    • Erkin Kuru
    • , Michael S. VanNieuwenhze
    •  & Yves V. Brun

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Contributions

C.G. and C.L. wrote the paper. A.F., C.L., S.M., C.Z., C.Cl., J.-P.L., M.F.-W. and C.G. designed and performed the experiments. C.G., C.L., A.F., S.M., J.-P.L., C.Co., M.F.-W. and B.M. analysed and interpreted the data. E.K., Y.V.B. and M.S.VN. provided new reagents. E.K., Y.B. and D.S. read and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Christian Lesterlin or Christophe Grangeasse.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Tables 1 and 2 and Supplementary References.

Videos

  1. 1.

    Time-lapse analysis of GFP-MapZ in wild-type cells

    Time-lapse analysis of GFP-MapZ in wild-type cells.

  2. 2.

    3D-SIM of FtsZ-GFP in wild-type cells

    3D-SIM of FtsZ-GFP in wild-type cells.

  3. 3.

    3D-SIM of FtsZ-GFP in ΔmapZ cells

    3D-SIM of FtsZ-GFP in ΔmapZ cells.

  4. 4.

    Time-lapse analysis of FtsZ-GFP in wild-type cells

    The video shows an overlay of GFP (green) and phase-contrast (gray) images.

  5. 5.

    Time-lapse analysis of FtsZ-GFP in ΔmapZ cells

    The video shows an overlay of GFP (green) and phase-contrast (gray) images.

  6. 6.

    3D-SIM of DAPI-stained nucleoids in ΔmapZ cells showing chromosome pinching

    3D-SIM of DAPI-stained nucleoids in ΔmapZ cells showing chromosome pinching

  7. 7.

    3D-SIM of FtsZ-GFP in DAPI-stained ΔmapZ cells showing aberrant chromosome associated with aberrant FtsZ structures

    3D-SIM of FtsZ-GFP in DAPI-stained ΔmapZ cells showing aberrant chromosome associated with aberrant FtsZ structures.

  8. 8.

    Time-lapse analysis of FtsZ-GFP in mapZΔ(1-41) cells

    The video shows an overlay of GFP (green) and phase-contrast (gray) images.

  9. 9.

    Time-lapse analysis of FtsZ-GFP in mapZΔcyto cells

    The video shows an overlay of GFP (green) and phase-contrast (gray) images.

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DOI

https://doi.org/10.1038/nature13966

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