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Oriented loading of FtsK on KOPS

Nature Structural & Molecular Biology volume 13, pages 1026–1028 (2006)Cite this article

Abstract

In Escherichia coli, the ATP-dependent DNA translocase FtsK transports DNA across the site of cell division and activates recombination by the XerCD recombinases at a specific site on the chromosome, dif, to ensure the last stages of chromosome segregation. DNA transport by FtsK is oriented by 8-base-pair asymmetric sequences ('KOPS'). Here we provide evidence that KOPS promote FtsK loading on DNA and that translocation is oriented at this step.

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Figure 1: KOPS are enhancers of FtsK activity.
Figure 2: DNA loop formation by FtsK preferentially occurs downstream of KOPS.
Figure 3: Model for oriented loading of FtsK on KOPS (arrowheads) and for translocation reversal when KOPS are met from a downstream position.

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References

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Acknowledgements

S.B. and O.A.S. were supported by a MENESR grant and an EU MolSwitch grant, respectively. Research was supported by the Centre national pour la recherche scientifique (ATIP), the French Ministry of Research (DRAB), the Fondation pour la Recherche Médicale and the Agence Nationale de la Recherche. We thank B. Michel, B. Lestrade, O. Espeli and C. Lesterlin for helpful comments.

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Authors and Affiliations

  1. Laboratoire de Microbiologie et de Génétique Moléculaire, CNRS, UMR5100, Toulouse, France

    Sarah Bigot & François Cornet

  2. Centre de Génétique Moléculaire, CNRS, UPR2167, Gif-sur-Yvette, France

    Sarah Bigot & François–Xavier Barre

  3. Laboratoire de Physique Statistique et Département de Biologie, Ecole Normale Supérieure, UMR8550 associé au CNRS et aux Universités Paris VI et VII, Paris, France

    Omar A Saleh & Jean-François Allemand

  4. Département de Chimie, Laboratoire Pasteur, Ecole Normale Supérieure, Paris, France

    Omar A Saleh & Jean-François Allemand

  5. Materials Department and Biomolecular Science and Engineering Program, University of California, Santa Barbara, California, USA

    Omar A Saleh

Authors
  1. Sarah Bigot
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  2. Omar A Saleh
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  3. François Cornet
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  4. Jean-François Allemand
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  5. François–Xavier Barre
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Contributions

DNA engineering, protein purification and bulk analysis were performed by S.B., F.C. and F.-X.B., and single-molecule studies by O.A.S. and J.-F.A.

Corresponding author

Correspondence to François–Xavier Barre.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Recombination on two dif sites in direct repeat. (PDF 567 kb)

Supplementary Fig. 2

Displacement of dif sites upsteam decreases their inhibitory effect on recombination. (PDF 242 kb)

Supplementary Fig. 3

Effect of KOPS on ATP hydrolysis. (PDF 97 kb)

Supplementary Fig. 4

Translocation velocity monitored in vitro by tracking real-time extension of a single DNA molecule. (PDF 279 kb)

Supplementary Methods (PDF 43 kb)

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Bigot, S., Saleh, O., Cornet, F. et al. Oriented loading of FtsK on KOPS. Nat Struct Mol Biol 13, 1026–1028 (2006). https://doi.org/10.1038/nsmb1159

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