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Detection of Protein SUMOylation In Situ by Proximity Ligation Assays

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SUMO

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1475))

Abstract

Sumoylation is a posttranslational process essential for life and concerns a growing number of crucial proteins. Understanding the influence of this phenomenon on individual proteins or on cellular pathways in which they function has become an intense area of research. A critical step in studying protein sumoylation is to detect sumoylated forms of a particular protein. This has proven to be a challenging task for a number of reasons, especially in the case of endogenous proteins and in vivo studies or when studying rare cells such as stem cells. Proximity ligation assays that allow detection of closely interacting protein partners can be adapted for initial detection of endogenous sumoylation or ubiquitination in a rapid, ultrasensitive, and cheap manner. In addition, modified forms of a given protein can be detected in situ in various cellular compartments. Finally, the flexibility of this technique may allow rapid screening of drugs and stress signals that may modulate protein sumoylation.

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Acknowledgements

We thank the core facilities of the IUH (Institut Universitaire d’Hématologie). We thank the technology platform at IUH, in particular Niclas Setterbald for his help with microscopy analysis. The laboratory is supported by INSERM, CNRS, Université Paris-Diderot, Collège de France, Ligue Contre le Cancer, Institut National du Cancer, the French National Research Agency (ANR) as part of the “Investissements d’Avenir” program (reference: ANR-11-PHUC-002 and ANR-13-JSV2-0005-01), Association pour la Recherche contre le Cancer (Griffuel Award to H. de Thé), Canceropôle Ile de France, the European Research Council (STEMAPL advanced grant to H. de Thé), PACRI and Saint Louis Institute U. Sahin was supported by a fellowship of the Fondation pour la Recherche Medicale. Authors’ contributions: F.J., C.B., and U.S. performed experiments; H.dT., U.S., and V.L.B. designed experiments, analyzed the data, and wrote the manuscript.

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Authors and Affiliations

  1. INSERM U944 - CNRS UMR7212 - Université Paris Diderot, Sorbonne Paris Cité - Equipe labellisée par la Ligue Nationale contre le Cancer, IUH, Hôpital St. Louis 1, Avenue Claude Vellefaux, 75475, Paris cedex 10, France

    Umut Sahin, Florence Jollivet, Caroline Berthier, Hugues de Thé & Valérie Lallemand-Breitenbach

  2. Department of Molecular Biology and Genetics, Center for Life Sciences and Technologies, Boğaziçi University, Istanbul, 34342, Turkey

    Umut Sahin

  3. Collège de France, University of Paris Science Lettre, Paris, France

    Caroline Berthier, Hugues de Thé & Valérie Lallemand-Breitenbach

  4. AP-HP, Service de Biochimie, Hôpital St. Louis, Paris, France

    Hugues de Thé

Authors
  1. Umut Sahin
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  2. Florence Jollivet
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  3. Caroline Berthier
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  4. Hugues de Thé
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  5. Valérie Lallemand-Breitenbach
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Corresponding authors

Correspondence to Umut Sahin or Valérie Lallemand-Breitenbach .

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Editors and Affiliations

  1. INBIOMED, San Sebastian, Spain

    Manuel S. Rodriguez

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© 2016 Springer Science+Business Media New York

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Sahin, U., Jollivet, F., Berthier, C., de Thé, H., Lallemand-Breitenbach, V. (2016). Detection of Protein SUMOylation In Situ by Proximity Ligation Assays. In: Rodriguez, M. (eds) SUMO. Methods in Molecular Biology, vol 1475. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6358-4_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6358-4_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6356-0

  • Online ISBN: 978-1-4939-6358-4

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