The permease gene nagE2 is the key to N-acetylglucosamine sensing and utilization in Streptomyces coelicolor and is subject to multi-level control

Authors

  • Harald Nothaft,

    1. Department of Biological Sciences and the Alberta Ingenuity Centre for Carbohydrate Science, University of Alberta, Edmonton, Alberta T6G 2R3, Canada.
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  • Sébastien Rigali,

    1. Centre for Protein Engineering, Université de Liège, Institut de Chimie B6a, Sart Tilman, B-4000 Liège, Belgium.
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  • Bart Boomsma,

    1. Molecular Systems Biotechnology, Leiden Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
    2. Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK.
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  • Magdalena Swiatek,

    1. Molecular Systems Biotechnology, Leiden Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
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  • Kenneth J. McDowall,

    1. Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK.
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  • Gilles P. Van Wezel,

    1. Molecular Systems Biotechnology, Leiden Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
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  • Fritz Titgemeyer

    Corresponding author
    1. Department of Oecotrophologie, Münster University of Applied Sciences, Corrensstr. 25, 48149 Münster, Germany.
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*E-mail titgemeyer@fh-muenster.de; Tel. (+49) 251 8365422; Fax (+49) 251 8365402.

Summary

The availability of nutrients is a major determinant for the timing of morphogenesis and antibiotic production in the soil-dwelling bacterium Streptomyces coelicolor. Here we show that N-acetylglucosamine transport, the first step of an important nutrient signalling cascade, is mediated by the NagE2 permease of the phosphotransferase system, and that the activity of this permease is linked to nutritional control of development and antibiotic production. The permease serves as a high-affinity transporter for N-acetylglucosamine (Km of 2.6 µM). The permease complex was reconstituted with individually purified components. This showed that uptake of N-acetylglucosamine requires a phosphoryl group transfer from phosphoenolpyruvate via the phosphotransferases EI, HPr and IIACrr to NagF, which in turn phosphorylates N-acetylglucosamine during transport. Transcription of the nagF and nagE2 genes is induced by N-acetylglucosamine. Nutrient signalling by N-acetylglucosamine that triggers the onset of development was abolished in the nagE2 and nagF mutants. nagE2 is subject to multi-level control by the global transcription factor DasR and the activator AtrA that also stimulates genes for antibiotic actinorhodin biosynthesis. Hence, it is apparent that streptomycetes tightly control the nutritional state in a complex manner to ensure the correct timing for the developmental programme.

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