Penicillin-binding protein folding is dependent on the PrsA peptidyl-prolyl cis-trans isomerase in Bacillus subtilis

Authors

  • Hanne-Leena Hyyryläinen,

    1. Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
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  • Bogumila C. Marciniak,

    1. Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN Haren, the Netherlands.
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  • Kathleen Dahncke,

    1. Institut für Mikrobiologie, Ernst-Moritz-Arndt Universität Greifswald, Friedrich-Ludwig-Jahn-Str. 15, D-17489 Greifswald, Germany.
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    • Present addresses: Abteilung Biochemie der Pflanzen, Institut für Biologie, Freie Universität Berlin, Königin-Luise-Straße 12-16, D-14195 Berlin, Germany;

  • Milla Pietiäinen,

    1. Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
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  • Pascal Courtin,

    1. INRA, UR477 Biochimie Bactérienne, F-78350 Jouy-en-Josas, France.
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  • Marika Vitikainen,

    1. Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
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    • VTT Technical Research Centre of Finland, Biotechnology, Tietotie 2, Espoo, P.O. Box 1000, FI-02044 VTT, Finland.

  • Raili Seppala,

    1. Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
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  • Andreas Otto,

    1. Institut für Mikrobiologie, Ernst-Moritz-Arndt Universität Greifswald, Friedrich-Ludwig-Jahn-Str. 15, D-17489 Greifswald, Germany.
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  • Dörte Becher,

    1. Institut für Mikrobiologie, Ernst-Moritz-Arndt Universität Greifswald, Friedrich-Ludwig-Jahn-Str. 15, D-17489 Greifswald, Germany.
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  • Marie-Pierre Chapot-Chartier,

    1. INRA, UR477 Biochimie Bactérienne, F-78350 Jouy-en-Josas, France.
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  • Oscar P. Kuipers,

    1. Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN Haren, the Netherlands.
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  • Vesa P. Kontinen

    Corresponding author
    1. Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
      E-mail vesa.kontinen@thl.fi; vesa.p.kontinen@helsinki.fi; Tel. (+358) 20 610 68562; Fax (+358) 20 610 68238.
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E-mail vesa.kontinen@thl.fi; vesa.p.kontinen@helsinki.fi; Tel. (+358) 20 610 68562; Fax (+358) 20 610 68238.

Summary

The PrsA protein is a membrane-anchored peptidyl-prolyl cis-trans isomerase in Bacillus subtilis and most other Gram-positive bacteria. It catalyses the post-translocational folding of exported proteins and is essential for normal growth of B. subtilis. We studied the mechanism behind this indispensability. We could construct a viable prsA null mutant in the presence of a high concentration of magnesium. Various changes in cell morphology in the absence of PrsA suggested that PrsA is involved in the biosynthesis of the cylindrical lateral wall. Consistently, four penicillin-binding proteins (PBP2a, PBP2b, PBP3 and PBP4) were unstable in the absence of PrsA, while muropeptide analysis revealed a 2% decrease in the peptidoglycan cross-linkage index. Misfolded PBP2a was detected in PrsA-depleted cells, indicating that PrsA is required for the folding of this PBP either directly or indirectly. Furthermore, strongly increased uniform staining of cell wall with a fluorescent vancomycin was observed in the absence of PrsA. We also demonstrated that PrsA is a dimeric or oligomeric protein which is localized at distinct spots organized in a helical pattern along the cell membrane. These results suggest that PrsA is essential for normal growth most probably as PBP folding is dependent on this PPIase.

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