Using FRET to Monitor Protein-Induced DNA Bending: The TBP-TATA Complex as a Model System

  • Rebecca H. Blair
  • James A. Goodrich
  • Jennifer F. Kugel
Part of the Methods in Molecular Biology book series (MIMB, volume 977)


Proteins that bind to DNA can elicit changes in DNA conformation, such as bending and looping, which are important signals for later events such as transcription. TATA-binding protein (TBP) is one example of a protein that elicits a conformational change in DNA; TBP binds and sharply bends its recognition sequence, which is thought to facilitate the recruitment of other protein factors. Here we describe the use of fluorescence resonance energy transfer (FRET) to evaluate DNA bending using TBP as a model system. FRET is a useful technique to measure changes in DNA conformation due to protein binding because small changes in the distance between two fluorophores (2–10 nm) translate into large changes in energy transfer.

Key words

FRET DNA bending Protein–DNA interaction TBP Fluorophore 



This work was supported by grant MCB-0919935 from the National Science Foundation.


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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Rebecca H. Blair
    • 1
  • James A. Goodrich
    • 1
  • Jennifer F. Kugel
    • 1
  1. 1.Department of Chemistry and BiochemistryUniversity of ColoradoBoulderUSA

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