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Functional Studies of DNA-Protein Interactions Using FRET Techniques

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DNA-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1334))

Abstract

Protein-DNA interactions underpin life and play key roles in all cellular processes and functions including DNA transcription, packaging, replication, and repair. Identifying and examining the nature of these interactions is therefore a crucial prerequisite to understand the molecular basis of how these fundamental processes take place. The application of fluorescence techniques and in particular fluorescence resonance energy transfer (FRET) to provide structural and kinetic information has experienced a stunning growth during the past decade. This has been mostly promoted by new advances in the preparation of dye-labeled nucleic acids and proteins and in optical sensitivity, where its implementation at the level of individual molecules has opened a new biophysical frontier. Nowadays, the application of FRET-based techniques to the analysis of protein-DNA interactions spans from the classical steady-state and time-resolved methods averaging over large ensembles to the analysis of distances, conformational changes, and enzymatic reactions in individual protein-DNA complexes. This chapter introduces the practical aspects of applying these methods for the study of protein-DNA interactions.

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Acknowledgments

We thank the Biological and Biotechnology Science Research Council (UK), the Royal Society (UK), and the National Sciences and Engineering Research Council (Canada) and the Universities of Sherbrooke (Canada) and St Andrews (UK) for financial support. We also thank all members of our labs for helpful discussion and critical reading of the manuscript. J.C.P. is a Fellow of the Scottish Universities Physics Alliance (SUPA).

Author information

Authors and Affiliations

  1. Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, QC, Canada

    Simon Blouin

  2. Department of Physics, Oxford University, Oxford, UK

    Timothy D. Craggs

  3. Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 Boulevard de L’Université, Sherbrooke, QC, Canada, J1K 2R1

    Daniel A. Lafontaine

  4. School of Physics and Astronomy, University of St. Andrews, St. Andrews, UK

    J. Carlos Penedo

Authors
  1. Simon Blouin
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  2. Timothy D. Craggs
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  3. Daniel A. Lafontaine
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  4. J. Carlos Penedo
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Corresponding author

Correspondence to Daniel A. Lafontaine .

Editor information

Editors and Affiliations

  1. Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada

    Benoît P. Leblanc

  2. Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada

    Sébastien Rodrigue

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Blouin, S., Craggs, T.D., Lafontaine, D.A., Penedo, J.C. (2015). Functional Studies of DNA-Protein Interactions Using FRET Techniques. In: Leblanc, B., Rodrigue, S. (eds) DNA-Protein Interactions. Methods in Molecular Biology, vol 1334. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2877-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-2877-4_8

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2876-7

  • Online ISBN: 978-1-4939-2877-4

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