Article | Published:

Intersubunit coordination in a homomeric ring ATPase

Nature volume 457, pages 446450 (22 January 2009) | Download Citation

Abstract

Homomeric ring ATPases perform many vital and varied tasks in the cell, ranging from chromosome segregation to protein degradation. Here we report the direct observation of the intersubunit coordination and step size of such a ring ATPase, the double-stranded-DNA packaging motor in the bacteriophage ϕ29. Using high-resolution optical tweezers, we find that packaging occurs in increments of 10 base pairs (bp). Statistical analysis of the preceding dwell times reveals that multiple ATPs bind during each dwell, and application of high force reveals that these 10-bp increments are composed of four 2.5-bp steps. These results indicate that the hydrolysis cycles of the individual subunits are highly coordinated by means of a mechanism novel for ring ATPases. Furthermore, a step size that is a non-integer number of base pairs demands new models for motor–DNA interactions.

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Acknowledgements

We thank C. L. Hetherington, M. Nollmann and G. Chistol for a critical reading of the manuscript; C. L. Hetherington, A. Politzer, M. Strycharska, M. Kopaczynska and J. Yu for critical discussions; and J. Choy, S. Grill and S. Smith for advice regarding instrumentation. J.R.M. acknowledges the National Science Foundation’s Graduate Research Fellowship and Y.R.C. the Burroughs Welcome Fund’s Career Awards at the Scientific Interface for funding. This research was supported in part by NIH grants GM-071552, DE-003606 and GM-059604. The content of this paper is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Author Contributions J.R.M., Y.R.C. and K.A. conducted the experiments and performed the analysis; S.G., P.J.J. and D.L.A. prepared and provided experimental materials; and J.R.M., Y.R.C., K.A., S.G., P.J.J., D.L.A. and C.B. wrote the paper. J.R.M. and Y.R.C. contributed equally to this work.

Author information

Author notes

    • Jeffrey R. Moffitt
    •  & Yann R. Chemla

    These authors contributed equally to this work.

    • Yann R. Chemla

    Present address: Department of Physics and Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.

Affiliations

  1. Department of Physics and Jason L. Choy Laboratory of Single Molecule Biophysics,

    • Jeffrey R. Moffitt
    • , Yann R. Chemla
    •  & Carlos Bustamante
  2. Biophysics Graduate Group, University of California, Berkeley, California 94720, USA

    • K. Aathavan
    •  & Carlos Bustamante
  3. Department of Diagnostic and Biological Sciences,

    • Shelley Grimes
    • , Paul J. Jardine
    •  & Dwight L. Anderson
  4. Department of Microbiology, University of Minnesota, Minneapolis, Minnesota 55455, USA

    • Dwight L. Anderson
  5. Departments of Molecular and Cell Biology, Chemistry, and Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA

    • Carlos Bustamante

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Correspondence to Carlos Bustamante.

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    Supplementary Information

    This file contains Supplementary Methods, a Supplementary Discussion, Supplementary Figures 1-8 with Legends and Supplementary References

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https://doi.org/10.1038/nature07637

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