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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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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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.
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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)
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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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DOI: https://doi.org/10.1038/nsmb1159