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A spindle-like apparatus guides bacterial chromosome segregation

Nature Cell Biology volume 12, pages 791–798 (2010)Cite this article

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Abstract

Until recently, a dedicated mitotic apparatus that segregates newly replicated chromosomes into daughter cells was believed to be unique to eukaryotic cells. Here we demonstrate that the bacterium Caulobacter crescentus segregates its chromosome using a partitioning (Par) apparatus that has surprising similarities to eukaryotic spindles. We show that the C. crescentus ATPase ParA forms linear polymers in vitro and assembles into a narrow linear structure in vivo. The centromere-binding protein ParB binds to and destabilizes ParA structures in vitro. We propose that this ParB-stimulated ParA depolymerization activity moves the centromere to the opposite cell pole through a burnt bridge Brownian ratchet mechanism. Finally, we identify the pole-specific TipN protein1,2 as a new component of the Par system that is required to maintain the directionality of DNA transfer towards the new cell pole. Our results elucidate a bacterial chromosome segregation mechanism that features basic operating principles similar to eukaryotic mitotic machines, including a multivalent protein complex at the centromere that stimulates the dynamic disassembly of polymers to move chromosomes into daughter compartments.

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Figure 1: ParA and ParB dynamics in vivo and ParA polymerization in vitro suggest a retracting polymeric ParA structure guides centromere segregation.
Figure 2: Mutational and biochemical analysis of C. crescentus ParA.
Figure 3: ParB in complex with parS drives the dynamics of ParA structures on DNA.
Figure 4: TipN confers new pole-specific directionality to Par-mediated DNA transfer through direct interaction with ParA.
Figure 5: A burnt-bridge Brownian ratchet mechanism for Par-mediated chromosome segregation in C. crescentus.

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Acknowledgements

We thank Jimmy Blair for assistance with modelling of ParA mutants, and critical reading of the manuscript; and Grant Bowman, Erin Goley and Julie Biteen for technical advice. We thank Jian Zhu and Thomas Earnest for providing purified 6His–ParB. This work is supported by National Institutes of Health grants R01 GM51426 R24 and GM073011-04d to L.S., NIH/NIGMS fellowship F32GM088966-1 to J.P., NIH/NIGMS award R01GM086196-2 to W.E.M., the Smith Stanford Graduate Fellowship to E.T., and a Helen Hay Whitney postdoctoral fellowship to E.G. This work was also supported by the Director, Office of Science, Office of Biological and Environmental Research, of the U.S. Department of Energy under contract no. DE-AC02-05CH11231.

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Authors and Affiliations

  1. Department of Developmental Biology, Stanford University School of Medicine, Beckman Center, Stanford, 94305, CA, USA

    Jerod L. Ptacin, Esteban Toro & Lucy Shapiro

  2. Department of Chemistry, Stanford University, Stanford, 94305, CA, USA

    Steven F. Lee & W.E. Moerner

  3. Department of Systems Biology, Harvard Medical School, Boston, 02115, MA, USA

    Ethan C. Garner

  4. Stanford Protein and Nucleic Acid Facility, Stanford University School of Medicine, Beckman Center, Stanford, 94305, CA, USA

    Michael Eckart

  5. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, 94720, CA, USA

    Luis R. Comolli

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  1. Jerod L. Ptacin
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Contributions

J.P., S.L., W.E.M. and L.S. designed the research; J.P. performed C. crescentus genetic, epifluorescence microscopy and biochemical experiments; S.L. performed single molecule imaging and data analysis; E.G. purified native ParA and performed ParA light-scattering experiments; E.T. designed ParA/DNA SPR experiments and performed time-lapse microscopy experiments on ΔtipN strains; M.E. performed SPR experiments and analysis; L.C. performed ParA negative-stain electron microscopy imaging; W.E.M. and L.S. supervised the study; J.P., S.L., W.E.M. and L.S. wrote the paper.

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Correspondence to Lucy Shapiro.

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Ptacin, J., Lee, S., Garner, E. et al. A spindle-like apparatus guides bacterial chromosome segregation. Nat Cell Biol 12, 791–798 (2010). https://doi.org/10.1038/ncb2083

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