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DNA transport into Bacillus subtilis requires proton motive force to generate large molecular forces

Nature Structural & Molecular Biology volume 11, pages 643–649 (2004)Cite this article

Abstract

Bacteria can acquire genetic diversity, including antibiotic resistance and virulence traits, by horizontal gene transfer. In particular, many bacteria are naturally competent for uptake of naked DNA from the environment in a process called transformation. Here, we used optical tweezers to demonstrate that the DNA transport machinery in Bacillus subtilis is a force-generating motor. Single DNA molecules were processively transported in a linear fashion without observable pausing events. Uncouplers inhibited DNA uptake immediately, suggesting that the transmembrane proton motive force is needed for DNA translocation. We found an uptake rate of 80 ± 10 bp s−1 that was force-independent at external forces <40 pN, indicating that a powerful molecular machine supports DNA transport.

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Figure 1: Molecular model for DNA transport through the cell envelope in Bacillus subtilis.
Figure 2: Experimental setup.
Figure 3: DNA binding.
Figure 4: Kinetics of DNA uptake.
Figure 5: Force generation during DNA transport.
Figure 6: DNA uptake is dependent on proton motive force.

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Acknowledgements

We thank J. Hahn for strain BD3773 and invaluable discussions. The project was supported by grant GM3677 and D.D. by grant GM43756. B.M. acknowledges support by the Deutsche Forschungsgemeinschaft (Emmy Noether Programm).

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Author notes

  1. Berenike Maier

    Present address: Institut für Experimentalphysik, Ludwig-Maximilians-Universität, Geschwister-Scholl Platz 1, 80539, München, Germany

Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, 1212 Amsterdam Ave., New York, 10027, New York, USA

    Berenike Maier & Michael P Sheetz

  2. Public Health Research Institute, 225 Warren Street, Newark, 07103, New Jersey, USA

    Ines Chen & David Dubnau

Authors
  1. Berenike Maier
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  2. Ines Chen
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Correspondence to Berenike Maier.

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Supplementary information

Supplementary Fig. 1

Example for the distance between the bead in the optical trap and the bacterium attached to the glass coverslide during DNA uptake. (PDF 372 kb)

Supplementary Fig. 2

Elasticity of DNA in competence medium. (PDF 84 kb)

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Maier, B., Chen, I., Dubnau, D. et al. DNA transport into Bacillus subtilis requires proton motive force to generate large molecular forces. Nat Struct Mol Biol 11, 643–649 (2004). https://doi.org/10.1038/nsmb783

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