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Regulation of the clpP1clpP2 operon by the pleiotropic regulator AdpA in Streptomyces lividans

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Abstract

Insertion of an apramycin resistance cassette in the clpP1clpP2 operon (encoding the ClpP1 and ClpP2 peptidase subunits) affects morphological and physiological differentiation of Streptomyces lividans. Another key factor controlling Streptomyces differentiation is the pleiotropic transcriptional regulator AdpA. We have identified a spontaneous missense mutation (−1 frameshift) in the adpA (bldH) open reading frame in a clpP1clpP2 mutant that led to the synthesis of a non-functional AdpA protein. Electrophoretic mobility shift assays showed that AdpA bound directly to clpP1clpP2 promoter region. Quantitative real-time PCR analysis showed that AdpA regulated the clpP1clpP2 operon expression at specific growth times. In vitro, AdpA and ClgR, a transcriptional activator of clpP1clpP2 operon and other genes, were able to bind simultaneously to clpP1 promoter, which suggests that AdpA binding to clpP1 promoter did not affect that of ClgR. This study allowed to uncover an interplay between the ClpP peptidases and AdpA in S. lividans.

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Acknowledgments

We thank T. Msadek and S. Dubrac for helpful discussions and critical reading of the manuscript, O. Poupel for assistance with qRT-PCR analysis, and C. Proux for transcriptome analysis. We thank Alex Edelman and Associates for correcting the manuscript. This work was supported by the research funds from the Institut Pasteur, the Centre National de Recherche Scientifique, and from the ERA-IB European grant. A. Guyet was the recipient of fellowships from the Ministère de l’Education Nationale, de la Recherche et de la Technologie, the Pasteur-Weizmann foundation, and the ERA-IB European grant.

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Authors and Affiliations

  1. Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK

    Aurélie Guyet

  2. Unité Biologie des Bactéries Pathogènes à Gram-Positif, and CNRS URA 2172, Institut Pasteur, 28 rue du Dr. Roux, 75015, Paris, France

    Myriam Gominet & Philippe Mazodier

  3. Unité Biologie des Spirochètes, Institut Pasteur, 28 rue du Docteur Roux, 75724, Paris Cedex 15, France

    Nadia Benaroudj

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  1. Aurélie Guyet
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  2. Myriam Gominet
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  4. Philippe Mazodier
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Correspondence to Philippe Mazodier.

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Communicated by Jean-Luc Pernodet.

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Guyet, A., Gominet, M., Benaroudj, N. et al. Regulation of the clpP1clpP2 operon by the pleiotropic regulator AdpA in Streptomyces lividans . Arch Microbiol 195, 831–841 (2013). https://doi.org/10.1007/s00203-013-0918-2

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