1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics
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Functional instability allows access to DNA in longer Transcription Activator-Like Effector (TALE) arrays

  1. Kathryn Geiger-Schuller
  2. Jaba Mitra
  3. Taekjip Ha
  4. Doug Barrick  Is a corresponding author
  1. Johns Hopkins University, United States
  2. University of Illinois Urbana-Champaign, United States
Research Article
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Cite this article as: eLife 2019;8:e38298 doi: 10.7554/eLife.38298

Abstract

Transcription activator-like effectors (TALEs) bind DNA through an array of tandem 34-residue repeats. How TALE repeat domains wrap around DNA, often extending more than 1.5 helical turns, without using external energy is not well understood. Here, we examine the kinetics of DNA binding of TALE arrays with varying numbers of identical repeats. Single molecule fluorescence analysis and deterministic modeling reveal conformational heterogeneity in both the free- and DNA-bound TALE arrays. Our findings, combined with previously identified partly folded states, indicate a TALE instability that is functionally important for DNA binding. For TALEs forming less than one superhelical turn around DNA, partly folded states inhibit DNA binding. In contrast, for TALEs forming more than one turn, partly folded states facilitate DNA binding, demonstrating a mode of 'functional instability' that facilitates macromolecular assembly. Increasing repeat number slows down interconversion between the various DNA-free and DNA-bound states.

Article and author information

Author details

  1. Kathryn Geiger-Schuller

    Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6705-0681
  2. Jaba Mitra

    Materials Science and Engineering, University of Illinois Urbana-Champaign, Urbana, United States
    Competing interests
    No competing interests declared.
  3. Taekjip Ha

    Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, United States
    Competing interests
    Taekjip Ha, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2195-6258
  4. Doug Barrick

    Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, United States
    For correspondence
    barrick@jhu.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7291-1389

Funding

National Institute of General Medical Sciences (T32-GM008403)

  • Kathryn Geiger-Schuller

National Institute of General Medical Sciences (R01-GM068462)

  • Doug Barrick

National Institute of General Medical Sciences (GM1129659)

  • Taekjip Ha

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

Reviewing Editor

  1. Antoine M van Oijen, University of Wollongong, Australia

Publication history

  1. Received: May 11, 2018
  2. Accepted: February 27, 2019
  3. Accepted Manuscript published: February 27, 2019 (version 1)
  4. Version of Record published: April 12, 2019 (version 2)

Copyright

© 2019, Geiger-Schuller et al.

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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