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Pyrones as bacterial signaling molecules

Nature Chemical Biology volume 9, pages 573–578 (2013)Cite this article

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Abstract

Bacteria communicate via small diffusible molecules and thereby mediate group-coordinated behavior, a process referred to as quorum sensing. The prototypical quorum sensing system found in Gram-negative bacteria consists of a LuxI-type autoinducer synthase that produces N-acyl homoserine lactones (AHLs) as signals and a LuxR-type receptor that detects the AHLs to control expression of specific genes. However, many proteobacteria have proteins with homology to LuxR receptors yet lack any cognate LuxI-like AHL synthase. Here we show that in the insect pathogen Photorhabdus luminescens the orphan LuxR-type receptor PluR detects endogenously produced α-pyrones that serve as signaling molecules at low nanomolar concentrations. Additionally, the ketosynthase PpyS was identified as pyrone synthase. Reconstitution of the entire system containing PluR, the PluR-target operon we termed pcf and PpyS in Escherichia coli demonstrated that the cell-cell communication circuit is portable. Our research thus deorphanizes a signaling system and suggests that additional modes of bacterial communication may await discovery.

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Figure 1: The orphan LuxR-like receptor PluR regulates expression of the pcfABCDEF operon.
Figure 2: PPY bioactivity and structure. HPLC/MS traces and PpcfA reporter activity of P. luminescens and E. coli culture fluids and pure PPYs.
Figure 3: Pyrone binding by PluR.
Figure 4: Heterologous reconstruction of the PpyS-PluR cell-cell communication circuit using Pcf-derived cell clumping as readout.
Figure 5: Model of PpyS-PluR signaling in P. luminescens.

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Acknowledgements

Work in the Heermann lab was financially supported by the Deutsche Forschungsgemeinschaft ((DFG) HE 5247/4-1 and SPP 1617). Work in the Bode lab was supported by the DFG (SPP 1617) and a European Research Council starting grant under grant agreement no. 311477. We thank K. Jung for many helpful scientific discussions; S. Scheu and P. Grün for excellent technical assistance; Q. Zhou for help with HPLC/MS analysis; E.A. Alonso and F.I. Nollmann for help with the PPY production kinetics, S. Kinski for help with ketosynthase mutant construction; and H. Janosz for generation of E. coli insect pathogenicity assays. We are grateful to M. Schobert (Braunschweig) for providing E. coli strain ST18, H. Jung (München) for providing plasmid pUC19-Kan and H. Schweizer (Fort Collins, Colorado) for providing the Tn7 plasmids.

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Author notes

  1. Alexander O Brachmann, Sophie Brameyer, Helge B Bode and Ralf Heermann: These authors contributed equally to this work.

Authors and Affiliations

  1. Merck Stiftungsprofessur für Molekulare Biotechnologie, Goethe-Universität Frankfurt, Frankfurt, Germany

    Alexander O Brachmann, Darko Kresovic, Yannick Kopp & Helge B Bode

  2. Biozentrum, Bereich Mikrobiologie, Ludwig-Maximilians-Universität München, München, Germany

    Sophie Brameyer, Ivana Hitkova, Christian Manske, Karin Schubert & Ralf Heermann

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Contributions

A.O.B. performed PPY isolation, structure elucidation and quantification and identified ppyS. S.B. performed qRT-PCR, analyzed and quantified PPY bioactivity and specificity for PluR, performed cell clumping assays, generated amino acid replacements in PluR and measured the influence on pyrone sensing. Y.K. constructed PPY-producing E. coli strains. D.K. modeled PluR structure and performed PPY docking and in silico mutagenesis. I.H. performed two-dimensional PAGE, northern blot analyses and Photorhabdus pathogenicity assays. C.M. characterized chemical properties of the PluR signal and performed first E. coli cell clumping assays. K.S. performed preliminary fractionations of P. luminescens supernatants via HPLC and analyzed them for PluR-signal presence. H.B.B. and R.H. coordinated and designed the experiments, analyzed the data and wrote the manuscript.

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Correspondence to Helge B Bode or Ralf Heermann.

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Brachmann, A., Brameyer, S., Kresovic, D. et al. Pyrones as bacterial signaling molecules. Nat Chem Biol 9, 573–578 (2013). https://doi.org/10.1038/nchembio.1295

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