Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal

  1. James Richard Fuller
  2. Phoebe A Rice  Is a corresponding author
  1. The University of Chicago, United States

Abstract

The transposition of bacteriophage Mu serves as a model system for understanding DDE transposases and integrases. All available structures of these enzymes at the end of the transposition reaction, including Mu, exhibit significant bends in the transposition target site DNA. Here we use Mu to investigate the ramifications of target DNA bending on the transposition reaction. Enhancing the flexibility of the target DNA or prebending it increases its affinity for transpososomes by over an order of magnitude and increases the overall reaction rate. This and FRET confirm that flexibility is interrogated early during the interaction between the transposase and a potential target site, which may be how other DNA binding proteins can steer selection of advantageous target sites. We also find that the conformation of the target DNA after strand transfer is involved in preventing accidental catalysis of the reverse reaction, as conditions that destabilize this conformation also trigger reversal.

Article and author information

Author details

  1. James Richard Fuller

    Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Phoebe A Rice

    Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, United States
    For correspondence
    price@uchicago.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3467-341X

Funding

National Institute of General Medical Sciences (GM101989)

  • James Richard Fuller
  • Phoebe A Rice

National Center for Research Resources (1S10RR026988-01)

  • James Richard Fuller
  • Phoebe A Rice

National Science Foundation (DMR1420709)

  • James Richard Fuller
  • Phoebe A Rice

National Institute of General Medical Sciences (GM007183)

  • James Richard Fuller
  • Phoebe A Rice

National Institute of General Medical Sciences (GM104397)

  • James Richard Fuller
  • Phoebe A Rice

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. David Sherratt, University of Oxford, United Kingdom

Publication history

  1. Received: September 23, 2016
  2. Accepted: February 7, 2017
  3. Accepted Manuscript published: February 8, 2017 (version 1)
  4. Accepted Manuscript updated: February 13, 2017 (version 2)
  5. Version of Record published: March 17, 2017 (version 3)

Copyright

© 2017, Fuller & Rice

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. James Richard Fuller
  2. Phoebe A Rice
(2017)
Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal
eLife 6:e21777.
https://doi.org/10.7554/eLife.21777

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