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Proteins containing photosynthetic reaction centre domains modulate FtsZ-based archaeal cell division

Nature Microbiology volume 9, pages 698–711 (2024)Cite this article

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Abstract

Cell division in all domains of life requires the orchestration of many proteins, but in Archaea most of the machinery remains poorly characterized. Here we investigate the FtsZ-based cell division mechanism in Haloferax volcanii and find proteins containing photosynthetic reaction centre (PRC) barrel domains that play an essential role in archaeal cell division. We rename these proteins cell division protein B 1 (CdpB1) and CdpB2. Depletions and deletions in their respective genes cause severe cell division defects, generating drastically enlarged cells. Fluorescence microscopy of tagged FtsZ1, FtsZ2 and SepF in CdpB1 and CdpB2 mutant strains revealed an unusually disordered divisome that is not organized into a distinct ring-like structure. Biochemical analysis shows that SepF forms a tripartite complex with CdpB1/2 and crystal structures suggest that these two proteins might form filaments, possibly aligning SepF and the FtsZ2 ring during cell division. Overall our results indicate that PRC-domain proteins play essential roles in FtsZ-based cell division in Archaea.

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Fig. 1: Cellular positioning of PRC-barrel proteins CdpB1 and CdpB2 and their co-localization with divisome protein SepF.
Fig. 2: Effect of CdpB1 depletion and CdpB2 deletion on cell shape and the positioning of cell division proteins.
Fig. 3: CdpB1 and CdpB2 localization during SepF depletion.
Fig. 4: Biochemical analysis of CdpB1, CdpB2 and SepF from A. fulgidus.
Fig. 5: Crystal structure of the CdpB1–CdpB2 complex and cryo-EM with SepF and liposomes.
Fig. 6: Model for CdpB1/B2 function in Euryarchaeota.

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Data availability

The data that support the findings of this study are available in the Source Data. Source data are provided with this paper. Coordinates have been submitted to the Protein Data Bank (PDB) with accession code 8QZO.

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Acknowledgements

We thank X. Ye (ISTA) for providing the His–SUMO expression plasmid pSVA13429. pCDB302 was a gift from C. Bahl (Addgene plasmid number 113673; http://n2t.net/addgene:113673; RRID Addgene_113673). We thank B. Ahsan, G. Sharov, G. Cannone and S. Chen from the Medical Research Council (MRC) LMB Electron Microscopy Facility for help and support. We thank Scientific Computing at the MRC LMB for their support. We thank L. Trübestein and N. Krasnici of the protein service unit of the ISTA Lab Support Facility for help with the SEC coupled with multi-angle light scattering experiments. We thank D. Grohmann and R. Reichelt from the Archaea Centre at the University of Regensburg for providing the P. furiosus cell material. P.N. and S.-V.A. were supported by a Momentum grant from the Volkswagen (VW) Foundation (grant number 94933). D.K.-C. and D.B. were supported by the VW Stiftung ‘Life?’ programme (to J.L.; grant number Az 96727) and by the MRC, as part of UK Research and Innovation (UKRI), MRC file reference number U105184326 (to J.L.). N.T. and S.G. acknowledge support from the French Government’s Investissement d’Avenir program, Laboratoire d’Excellence ‘Integrative Biology of Emerging Infectious Diseases’ (grant number ANR-10-LABX-62-IBEID), and the computational and storage services (Maestro cluster) provided by the IT department at Institut Pasteur. M.K. and M.L. were supported by the Austrian Science Fund (FWF) Stand-Alone P34607. For the purpose of open access, the MRC Laboratory of Molecular Biology has applied a CC BY public copyright licence to any author accepted manuscript version arising.

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Authors and Affiliations

  1. Molecular Biology of Archaea, Microbiology, Faculty of Biology, University of Freiburg, Freiburg, Germany

    Phillip Nußbaum, Chris van der Does, Maxime Tourte & Sonja-Verena Albers

  2. MRC Laboratory of Molecular Biology, Cambridge, UK

    Danguole Kureisaite-Ciziene, Dom Bellini, Anna Yeates & Jan Löwe

  3. Institute of Science and Technology Austria (ISTA), Klosterneuburg, Austria

    Marko Kojic & Martin Loose

  4. Evolutionary Biology of the Microbial Cell Laboratory, Institut Pasteur, Université Paris Cité, Paris, France

    Najwa Taib & Simonetta Gribaldo

  5. Bioinformatics and Biostatistics Hub, Institut Pasteur, Université Paris Cité, Paris, France

    Najwa Taib

  6. Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany

    Sonja-Verena Albers

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Contributions

P.N. conceived the project and performed all experiments not otherwise mentioned. C.v.d.D. purified the proteins from A. fulgidus and performed SEC experiments. M.K. expressed and purified proteins from H. volcanii and performed SEC coupled with multi-angle light scattering experiments, pelleting assays and mass photometry. D.K.-C. and D.B. solved the crystal structure. D.K.-C., A.Y. and J.L. performed cryo-EM. N.T. performed phylogenetic analysis. M.T. isolated P. furiosus lipids. P.N. and J.L. prepared figures. P.N. wrote the draft of the article. S.G., M.L., J.L. and S.-V.A. reviewed drafts of the article, supervised the work and acquired funding.

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Correspondence to Sonja-Verena Albers.

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Nature Microbiology thanks Daniela Barilla, William Margolin 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. 1–13 and Tables 1–6.

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Supplementary Video 1

Time lapse of H. volcanii expressing CdpB1–mNeonGreen.

Supplementary Video 2

Time lapse of H. volcanii expressing CdpB2–mNeonGreen.

Supplementary Video 3

Time lapse of H. volcanii expressing CdpB3–mNeonGreen.

Supplementary Video 4

Time lapse of H. volcanii during CdpB1 depletion.

Supplementary Video 5

Time lapse of H. volcanii wild type.

Supplementary Data 1

Source data for supplementary figures.

Source data

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Nußbaum, P., Kureisaite-Ciziene, D., Bellini, D. et al. Proteins containing photosynthetic reaction centre domains modulate FtsZ-based archaeal cell division. Nat Microbiol 9, 698–711 (2024). https://doi.org/10.1038/s41564-024-01600-5

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