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Fusion Reporter Approaches to Monitoring Transmembrane Helix Interactions in Bacterial Membranes

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Bacterial Protein Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1615))

Abstract

In transenvelope multiprotein machines such as bacterial secretion systems, protein–protein interactions not only occur between soluble domains but might also be mediated by helix–helix contacts in the inner membrane. Here we describe genetic assays commonly used to test interactions between transmembrane α-helices in their native membrane environment. These assays are based on the reconstitution of dimeric regulators allowing the control of expression of reporter genes. We provide detailed protocols for the TOXCAT and GALLEX assays used to monitor homotypic and heterotypic transmembrane helix–helix interactions.

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Acknowledgements

Work in the EC laboratory is supported by the Centre National de la Recherche Scientifique, the Aix-Marseille Université, and grants from the Agence Nationale de la Recherche (ANR-14-CE14-0006-02 and ANR-15-CE11-0019-01). LL and AZ are recipients of doctoral fellowships from the French Ministère de l’Enseignement Supérieur et de la Recherche and end-of-thesis fellowships from the Fondation pour la Recherche Médicale (FDT20160435498 and FDT20140931060).

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Authors and Affiliations

  1. Laboratoire d’Ingénierie des Systèmes Macromoléculaires, UMR7255, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ—CNRS, 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France

    Laureen Logger, Abdelrahim Zoued & Eric Cascales

  2. Division of Infectious Diseases and Harvard Medical School, Department of Microbiology and Immunobiology, Howard Hughes Medical Institute, Brigham and Women’s Hospital, Boston, MA, USA

    Abdelrahim Zoued

Authors
  1. Laureen Logger
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  2. Abdelrahim Zoued
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  3. Eric Cascales
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Correspondence to Eric Cascales .

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Editors and Affiliations

  1. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Laure Journet

  2. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Eric Cascales

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Logger, L., Zoued, A., Cascales, E. (2017). Fusion Reporter Approaches to Monitoring Transmembrane Helix Interactions in Bacterial Membranes. In: Journet, L., Cascales, E. (eds) Bacterial Protein Secretion Systems. Methods in Molecular Biology, vol 1615. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7033-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7033-9_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7031-5

  • Online ISBN: 978-1-4939-7033-9

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