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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 543))

Summary

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). DAL is a CIHR New Investigator scholar.

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  1. Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 boul. Université, Sherbrooke, QC, Canada, J1K 2R1

    Simon Blouin, Timothy D. Craggs, Daniel A. Lafontaine & J. Carlos Penedo

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  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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Correspondence to J. Carlos Penedo .

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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Blouin, S., Craggs, T.D., Lafontaine, D.A., Penedo, J.C. (2009). Functional Studies of DNA-Protein Interactions Using FRET Techniques. In: Leblanc, B., Moss, T. (eds) DNA-Protein Interactions. Methods in Molecular Biology™, vol 543. Humana Press. https://doi.org/10.1007/978-1-60327-015-1_28

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    • DOI: https://doi.org/10.1007/978-1-60327-015-1_28

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