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A topological mechanism for TRF2-enhanced strand invasion

Nature Structural & Molecular Biology volume 14, pages 147–154 (2007)Cite this article

Abstract

Telomeres can fold into t-loops that may result from the invasion of the 3′ overhang into duplex DNA. Their formation is facilitated in vitro by the telomeric protein TRF2, but very little is known regarding the mechanisms involved. Here we reveal that TRF2 generates positive supercoiling and condenses DNA. Using a variety of TRF2 mutants, we demonstrate a strong correlation between this topological activity and the ability to stimulate strand invasion. We also report that these properties require the combination of the TRF-homology (TRFH) domain of TRF2 with either its N- or C-terminal DNA-binding domains. We propose that TRF2 complexes, by constraining DNA around themselves in a right-handed conformation, can induce untwisting of the neighboring DNA, thereby favoring strand invasion. Implications of this topological model in t-loop formation and telomere homeostasis are discussed.

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Figure 1: TRF2 enhances telomeric strand invasion.
Figure 2: RPA and POT1 binding do not impair strand invasion.
Figure 3: TRF2 modifies DNA topology.
Figure 4: TRF2 stimulates invasion outside the telomeric tract.
Figure 5: TRF2 condenses telomeric DNA.
Figure 6: A topological model for telomeric strand invasion.

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Acknowledgements

We thank R. Rahmouni for advice. This work was supported by grants from the Ligue Nationale contre le Cancer (“équipe labellisée”) and from “Région Centre”. C.L. is supported by fellowships from the Association pour la Recherche sur le Cancer.

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Authors and Affiliations

  1. Laboratoire de Biologie Moléculaire de la Cellule de l'Ecole Normale Supérieure de Lyon, CNRS UMR 5161, IFR128, 46 Allée d'Italie, Lyon, 69364, Cedex 07, France

    Simon Amiard, Sébastien Pinte, Anaïs Poulet, Christelle Lenain, Dimitar Angelov, Philippe Bouvet, Eric Gilson & Marie-Josèphe Giraud-Panis

  2. Centre de Biophysique Moléculaire, UPR301 CNRS, rue Charles Sadron, Orléans, 45071, Cedex 2, France

    Simon Amiard, Michel Doudeau & Jacques Paoletti

  3. Laboratoire Joliot Curie, Ecole Normale Supérieure, 46 Allée d'Italie, Lyon, 69364, Cedex 07, France

    Anaïs Poulet, Cendrine Faivre-Moskalenko, Dimitar Angelov, Philippe Bouvet & Marie-Josèphe Giraud-Panis

  4. Laboratoire de Physique, Ecole Normale Supérieure, 46 Allée d'Italie, Lyon, 69364, Cedex 07, France

    Cendrine Faivre-Moskalenko

  5. Swiss Institute for Experimental Cancer Research, Chemin des Boveresses 155, Epalinges s/Lausanne, CH-1066, Switzerland

    Nele Hug

  6. International Centre for Genetic Engineering and Biotechnology, Padriciano 99, Trieste, I-34012, Italy

    Alessandro Vindigni

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Correspondence to Eric Gilson.

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Supplementary information

Supplementary Fig. 1

Characterization of the telomeric strand invasion reaction. (PDF 144 kb)

Supplementary Fig. 2

DNA-binding properties of the TRF2 mutant forms used in this study. (PDF 223 kb)

Supplementary Fig. 3

Characterization of the TRF2-mediated strand invasion reaction. (PDF 109 kb)

Supplementary Fig. 4

TRF2 does not behave as a RecA-type protein. (PDF 101 kb)

Supplementary Fig. 5

TRF2, TRF2ΔB and TRF2ΔM form DNA complexes that do not migrate on polyacrylamide gels. (PDF 68 kb)

Supplementary Fig. 6

Topoisomers obtained in the topological assay do not originate from contaminations in the protein preparation. (PDF 65 kb)

Supplementary Fig. 7

TRF1, TRF2DM, TRF2DB and TRF2 do not create the same amount of supercoiling. (PDF 37 kb)

Supplementary Methods (PDF 103 kb)

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Amiard, S., Doudeau, M., Pinte, S. et al. A topological mechanism for TRF2-enhanced strand invasion. Nat Struct Mol Biol 14, 147–154 (2007). https://doi.org/10.1038/nsmb1192

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