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Analysis of DNA–Protein Complexes by Atomic Force Microscopy Imaging: The Case of TRF2–Telomeric DNA Wrapping

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Atomic Force Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1886))

Abstract

Atomic force microscopy (AFM) is a non-optical microscopy that enables the acquisition at the nanoscale level of a 3D topographical image of the sample. For 30 years, AFM has been a valuable tool in life sciences to study biological samples in the field of tissue, cellular and molecular imaging, of mechanical properties and of force spectroscopy. Since the early beginnings of the technique, AFM has been extensively exploited as an imaging tool for structural studies of nucleic acids and nucleoprotein complexes. The morphometric analysis performed on the images can unveil specific structural and functional aspects of the sample, such as the multimerization state of proteins bound to DNA, or DNA conformational changes led by the DNA-binding proteins. Herein, a method for analyzing a complex formed by a telomeric DNA sequence wrapped around the TRF2 binding protein is presented. The described procedure could be applied to the study of any type of DNA–protein complex.

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Acknowledgments

This work was funded by the Ligue Nationale contre le Cancer (Equipe labélisée) and Investments for the Future LABEXSIGNALIFE (reference ANR-236 11-LABX-0028-01).

The authors acknowledge PICMI, the IRCAN’s Imaging core facility. The atomic force microscopy of PICMI was supported by the Association pour la Recherche sur le Cancer (ARC), by the Infrastructures en Biologie Santé et Agronomie (IBiSA) and by the Conseil General 06 de la Région Provence Alpes-Côte.

Author information

Authors and Affiliations

  1. Université Côte d’Azur, CNRS UMR 7284/INSERM U108, Institute for Research on Cancer and Aging, Nice (IRCAN), Medical School, Nice, France

    Sabrina Pisano & Eric Gilson

  2. International Laboratory in Hematology and Cancer, Pôle Sino-Français de Recherche en Sciences du Vivant et Génomique, Shanghai Ruijin Hospital, Shanghai Jiao Tong University School of Medicine/Ruijin Hospital/CNRS/INSERM/Nice University, Shanghai, China

    Eric Gilson

  3. Department of Genetics, CHU Nice, Université Côte d’Azur, Nice, France

    Eric Gilson

Authors
  1. Sabrina Pisano
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  2. Eric Gilson
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Correspondence to Sabrina Pisano .

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Editors and Affiliations

  1. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal

    Nuno C. Santos

  2. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal

    Filomena A. Carvalho

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Pisano, S., Gilson, E. (2019). Analysis of DNA–Protein Complexes by Atomic Force Microscopy Imaging: The Case of TRF2–Telomeric DNA Wrapping. In: Santos, N., Carvalho, F. (eds) Atomic Force Microscopy. Methods in Molecular Biology, vol 1886. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8894-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-8894-5_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8893-8

  • Online ISBN: 978-1-4939-8894-5

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