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Surface-attached molecules control Staphylococcus aureus quorum sensing and biofilm development

Nature Microbiology volume 2, Article number: 17080 (2017) Cite this article

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Abstract

Bacteria use a process called quorum sensing to communicate and orchestrate collective behaviours, including virulence factor secretion and biofilm formation. Quorum sensing relies on the production, release, accumulation and population-wide detection of signal molecules called autoinducers. Here, we develop concepts to coat surfaces with quorum-sensing-manipulation molecules as a method to control collective behaviours. We probe this strategy using Staphylococcus aureus. Pro- and anti-quorum-sensing molecules can be covalently attached to surfaces using click chemistry, where they retain their abilities to influence bacterial behaviours. We investigate key features of the compounds, linkers and surfaces necessary to appropriately position molecules to interact with cognate receptors and the ability of modified surfaces to resist long-term storage, repeated infections, host plasma components and flow-generated stresses. Our studies highlight how this surface approach can be used to make colonization-resistant materials against S. aureus and other pathogens and how the approach can be adapted to promote beneficial behaviours of bacteria on surfaces.

Bacteria frequently act as members of collectives to create surface-attached communities called biofilms and to activate the production of virulence factors1–5. These and other bacterial group behaviours are regulated by a cell–cell chemical communication process called quorum sensing, which involves the production, release and population-wide detection of extracellular signal molecules called autoinducers6–8. There is significant interest in developing synthetic strategies to manipulate quorum sensing. Potential applications include the prevention of biofilm formation to reduce biofouling in industry and halt recurrent disease in medicine, and the suppression of virulence factor production to limit pathogen infectivity9–11. There is also interest in enhancing quorum-sensing-controlled behaviours in beneficial bacteria. For example, bacterial biofilm formation is essential in wastewater treatment and food processing and, in the context of the microbiome, commensal bacteria play roles in fending off invading pathogens, presumably by relying on quorum-sensing-controlled traits12,13. Indeed, anti- and pro-quorum-sensing compounds have been developed and show promise in relevant model systems14–17.

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Figure 1: A strategy to quantify the Agr quorum-sensing responses of S. aureus to exogenously supplied agonists and antagonists.
Figure 2: S. aureus Agr quorum sensing is activated by surface-attached AIP-I.
Figure 3: A surface-attached Agr quorum-sensing antagonist, TrAIP-II, inhibits the S. aureus quorum-sensing response to AIP-I.
Figure 4: Wild-type S. aureus responds to surface-attached AIP-I and TrAIP-II.
Figure 5: Features of autoinducer-attached surfaces.

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Acknowledgements

The authors acknowledge the Novick laboratory for providing S. aureus strains and plasmids. The authors thank N. Wingreen for discussions about heterogeniety, J. Yan for image analysis, B. Wang for discussion and mentoring in S. aureus genetic techniques, B. Bratton for discussions about single-molecule microscopy, I. Pelczer and K. Conover for the NMR measurements and D. Dabbs for help with the FTIR measurement. The authors acknowledge members of the B.L.B., H.A.S. and T.W.M. laboratories for suggestions. This work was supported by NSF grant MCB-1344191 (to B.L.B. and H.A.S.), the Howard Hughes Medical Institute, NIH grant 2R37GM065859 and NSF grant MCB-0948112 (to B.L.B.), NIH grant R01 AI042783 (to T.W.M.) and a STX fellowship (to M.K.K.).

Author information

Author notes

  1. Minyoung Kevin Kim and Aishan Zhao: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, Princeton University, Princeton, 08544, New Jersey, USA

    Minyoung Kevin Kim, Aishan Zhao, Ashley Wang, Zachary Z. Brown & Tom W. Muir

  2. Department of Mechanical and Aerospace Engineering, Princeton University, Princeton, 08544, New Jersey, USA

    Howard A. Stone

  3. Department of Molecular Biology, Princeton University, Princeton, 08544, New Jersey, USA

    Bonnie L. Bassler

  4. Howard Hughes Medical Institute, Chevy Chase, 20815, Maryland, USA

    Bonnie L. Bassler

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Contributions

M.K.K., H.A.S. and B.L.B. conceived the idea. M.K.K., A.Z., T.W.M., H.A.S. and B.L.B. designed the experiments. M.K.K. and A.Z. performed the majority of the experiments. A.W. helped with the solution assay. M.K.K., A.Z., A.W. and Z.Z.B. contributed new reagents/analytic tools. M.K.K., A.Z., T.W.M., H.A.S. and B.L.B. analysed the data. M.K.K., A.Z., H.A.S. and B.L.B. wrote the manuscript.

Corresponding authors

Correspondence to Howard A. Stone or Bonnie L. Bassler.

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The authors declare no competing financial interests.

Supplementary information

Supplementary information

Supplementary Figures 1–8; Supplementary Tables 1 and 2; Supplementary Note 1; Supplementary Methods; Supplementary References; colour-blind-friendly versions of Figures 1–5 and Supplementary Figures 1–8. (PDF 6644 kb)

Supplementary Video 1

S. aureus Agr quorum sensing is activated by surface attached AIP-I. Time series of merged fluorescence images of the S. Aureus reporter strain in the presence of Surface-PEG10000-triazole-AIP-I (top left), Surface-PEG10000-triazole-AIP-I + 2.5 μM TrAIP-II in solution (top right), Surface-PEG10000-azide (bottom left), and Surface-PEG10000-triazole LinearAIP-I (bottom right). The time interval between each image is 30 min. (AVI 2147 kb)

Supplementary Video 2

S. aureus Agr quorum sensing is inhibited by surface attached TrAIP-II. Time series of merged fluorescence images of the S. Aureus reporter strain in the presence of Surface-PEG10000-triazole-TrAIP-II + 30 nM AIP-I in solution (top left), SurfacePEG10000-triazole-TrAIP-II + 1 μM AIP-I in solution (top right), Surface-PEG10000-azide + 30 nM AIP-I in solution (bottom left), and Surface-PEG10000-triazole-Linear-TrAIP-II + 30 nM AIP-I in solution (bottom right). The time interval between each image is 30 min. (AVI 1750 kb)

Supplementary Video 3

Wild-type S. aureus responds to surface-attached AIPI and TrAIP-II. Time series of merged fluorescence images of wild-type S. aureus strain in the presence of Surface-PEG10000-azide (top left), SurfacePEG10000-triazole-AIP-I (top right), Surface-PEG10000-triazole-TrAIP-II (bottom left), and Surface-PEG10000-triazole-TrAIP-II + 1 μM AIP-I in solution (bottom right). The time interval between each image is 30 min. (AVI 1920 kb)

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Kim, M., Zhao, A., Wang, A. et al. Surface-attached molecules control Staphylococcus aureus quorum sensing and biofilm development. Nat Microbiol 2, 17080 (2017). https://doi.org/10.1038/nmicrobiol.2017.80

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