Bacterial quorum-sensing autoinducers are small chemicals released to control microbial community behaviours. N-(3-oxo-dodecanoyl) homoserine lactone, the autoinducer of the Pseudomonas aeruginosa LasI–LasR circuitry, triggers significant cell death in lymphocytes. We found that this molecule is incorporated into the mammalian plasma membrane and induces dissolution of eukaryotic lipid domains. This event expels tumour necrosis factor receptor 1 into the disordered lipid phase for its spontaneous trimerization without its ligand and drives caspase 3–caspase 8-mediated apoptosis. In vivo, P.aeruginosa releases N-(3-oxo-dodecanoyl) homoserine lactone to suppress host immunity for its own better survival; conversely, blockage of caspases strongly reduces the severity of the infection. This work reveals an unknown communication method between microorganisms and the mammalian host and suggests interventions of bacterial infections by intercepting quorum-sensing signalling.

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The data that support the findings of this study are available from the corresponding author upon request. Complete western blot images of all figures in the manuscript are provided as Supplementary Figures.

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  1. 1.

    Schuster, M., Sexton, D. J., Diggle, S. P. & Greenberg, E. P. Acyl-homoserine lactone quorum sensing: from evolution to application. Annu. Rev. Microbiol. 67, 43–63 (2013).

  2. 2.

    Smith, E. E. et al. Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients. Proc. Natl Acad. Sci. USA 103, 8487–8492 (2006).

  3. 3.

    McManus, A. T., Mason, A. D. Jr, McManus, W. F. & Pruitt, B. A. Jr. Twenty-five year review of Pseudomonas aeruginosa bacteremia in a burn center. Eur. J. Clin. Microbiol. 4, 219–223 (1985).

  4. 4.

    Pearson, J. P., Passador, L., Iglewski, B. H. & Greenberg, E. P. A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa. Proc. Natl Acad. Sci. USA 92, 1490–1494 (1995).

  5. 5.

    Davies, D. G. et al. The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280, 295–298 (1998).

  6. 6.

    Wagner, C. et al. The quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12-HSL) enhances the host defence by activating human polymorphonuclear neutrophils (PMN). Anal. Bioanal. Chem. 387, 481–487 (2007).

  7. 7.

    Tateda, K. et al. The Pseudomonas aeruginosa autoinducer N-3-oxododecanoyl homoserine lactone accelerates apoptosis in macrophages and neutrophils. Infect. Immun. 71, 5785–5793 (2003).

  8. 8.

    Skindersoe, M. E. et al. Pseudomonas aeruginosa quorum-sensing signal molecules interfere with dendritic cell-induced T-cell proliferation. FEMS Immunol. Med. Microbiol. 55, 335–345 (2009).

  9. 9.

    Glucksam-Galnoy, Y. et al. The bacterial quorum-sensing signal molecule N-3-oxo-dodecanoyl-l-homoserine lactone reciprocally modulates pro- and anti-inflammatory cytokines in activated macrophages. J. Immunol. 191, 337–344 (2013).

  10. 10.

    Smith, R. S., Harris, S. G., Phipps, R. & Iglewski, B. The Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)homoserine lactone contributes to virulence and induces inflammation in vivo. J. Bacteriol. 184, 1132–1139 (2002).

  11. 11.

    Kravchenko, V. V. et al. Modulation of gene expression via disruption of NF-κB signaling by a bacterial small molecule. Science 321, 259–263 (2008).

  12. 12.

    Jacobi, C. A. et al. Effects of bacterial N-acyl homoserine lactones on human Jurkat T lymphocytes-OdDHL induces apoptosis via the mitochondrial pathway. Int. J. Med. Microbiol. 299, 509–519 (2009).

  13. 13.

    Kravchenko, V. V. et al. N-(3-oxo-acyl)homoserine lactones signal cell activation through a mechanism distinct from the canonical pathogen-associated molecular pattern recognition receptor pathways. J. Biol. Chem. 281, 28822–28830 (2006).

  14. 14.

    Davis, B. M., Jensen, R., Williams, P. & O’Shea, P. The interaction of N-acylhomoserine lactone quorum sensing signaling molecules with biological membranes: implications for inter-kingdom signaling. PLoS ONE 5, e13522 (2010).

  15. 15.

    Favre-Bonte, S., Chamot, E., Kohler, T., Romand, J. A. & van Delden, C. Autoinducer production and quorum-sensing dependent phenotypes of Pseudomonas aeruginosa vary according to isolation site during colonization of intubated patients. BMC Microbiol. 7, 33 (2007).

  16. 16.

    Chhabra, S. R. et al. Synthetic analogues of the bacterial signal (quorum sensing) molecule N-(3-oxododecanoyl)-l-homoserine lactone as immune modulators. J. Med. Chem. 46, 97–104 (2003).

  17. 17.

    Charlton, T. S. et al. A novel and sensitive method for the quantification of N-3-oxoacyl homoserine lactones using gas chromatography–mass spectrometry: application to a model bacterial biofilm. Environ. Microbiol. 2, 530–541 (2000).

  18. 18.

    Erickson, D. L. et al. Pseudomonas aeruginosa quorum-sensing systems may control virulence factor expression in the lungs of patients with cystic fibrosis. Infect. Immun. 70, 1783–1790 (2002).

  19. 19.

    Hu, X. Proteolytic signaling by TNFα: caspase activation and IκB degradation. Cytokine 21, 286–294 (2003).

  20. 20.

    Atkinson, S., Chang, C. Y., Sockett, R. E., Camara, M. & Williams, P. Quorum sensing in Yersinia enterocolitica controls swimming and swarming motility. J. Bacteriol. 188, 1451–1461 (2006).

  21. 21.

    Barnhart, B. C., Alappat, E. C. & Peter, M. E. The CD95 type I/type II model. Semin. Immunol. 15, 185–193 (2003).

  22. 22.

    Lavrik, I. N. & Krammer, P. H. Regulation of CD95/Fas signaling at the DISC. Cell Death Differ. 19, 36–41 (2012).

  23. 23.

    Watanabe, N. et al. Continuous internalization of tumor necrosis factor receptors in a human myosarcoma cell line. J. Biol. Chem. 263, 10262–10266 (1988).

  24. 24.

    Shiner, E. K. et al. Pseudomonas aeruginosa autoinducer modulates host cell responses through calcium signalling. Cell. Microbiol. 8, 1601–1610 (2006).

  25. 25.

    Smith, R. S. et al. IL-8 production in human lung fibroblasts and epithelial cells activated by the Pseudomonas autoinducer N-3-oxododecanoyl homoserine lactone is transcriptionally regulated by NF-κB and activator protein-2. J. Immunol. 167, 366–374 (2001).

  26. 26.

    Jahoor, A. et al. Peroxisome proliferator-activated receptors mediate host cell proinflammatory responses to Pseudomonas aeruginosa autoinducer. J. Bacteriol. 190, 4408–4415 (2008).

  27. 27.

    Valentine, C. D., Anderson, M. O., Papa, F. R. & Haggie, P. M. X-box binding protein 1 (XBP1s) is a critical determinant of Pseudomonas aeruginosa homoserine lactone-mediated apoptosis. PLoS Pathog. 9, e1003576 (2013).

  28. 28.

    Zhu, J., Chai, Y., Zhong, Z., Li, S. & Winans, S. C. Agrobacterium bioassay strain for ultrasensitive detection of N-acylhomoserine lactone-type quorum-sensing molecules: detection of autoinducers in Mesorhizobium huakuii. Appl. Environ. Microbiol. 69, 6949–6953 (2003).

  29. 29.

    Schauder, S. & Bassler, B. L. The languages of bacteria. Genes Dev. 15, 1468–1480 (2001).

  30. 30.

    Legler, D. F., Micheau, O., Doucey, M. A., Tschopp, J. & Bron, C. Recruitment of TNF receptor 1 to lipid rafts is essential for TNFα-mediated NF-κB activation. Immunity 18, 655–664 (2003).

  31. 31.

    Muppidi, J. R., Tschopp, J. & Siegel, R. M. Life and death decisions: secondary complexes and lipid rafts in TNF receptor family signal transduction. Immunity 21, 461–465 (2004).

  32. 32.

    Lambert, W., Soderberg, C. A., Rutsdottir, G., Boelens, W. C. & Emanuelsson, C. Thiol-exchange in DTSSP crosslinked peptides is proportional to cysteine content and precisely controlled in crosslink detection by two-step LC–MALDI MSMS. Protein Sci. 20, 1682–1691 (2011).

  33. 33.

    Ulbrich, M. H. & Isacoff, E. Y. Subunit counting in membrane-bound proteins. Nat. Methods 4, 319–321 (2007).

  34. 34.

    Connell, S. D., Heath, G., Olmsted, P. D. & Kisil, A. Critical point fluctuations in supported lipid membranes. Faraday Discuss. 161, 91–111; discussion 113–150 (2013).

  35. 35.

    Rinia, H. A. & de Kruijff, B. Imaging domains in model membranes with atomic force microscopy. FEBS Lett. 504, 194–199 (2001).

  36. 36.

    Simons, K. & Vaz, W. L. Model systems, lipid rafts, and cell membranes. Annu. Rev. Biophys. Biomol. Struct. 33, 269–295 (2004).

  37. 37.

    Yuan, C., Furlong, J., Burgos, P. & Johnston, L. J. The size of lipid rafts: an atomic force microscopy study of ganglioside GM1 domains in sphingomyelin/DOPC/cholesterol membranes. Biophys. J. 82, 2526–2535 (2002).

  38. 38.

    Sengupta, P., Hammond, A., Holowka, D. & Baird, B. Structural determinants for partitioning of lipids and proteins between coexisting fluid phases in giant plasma membrane vesicles. Biochim. Biophys. Acta 1778, 20–32 (2008).

  39. 39.

    Fujiwara, T. K. et al. Confined diffusion of transmembrane proteins and lipids induced by the same actin meshwork lining the plasma membrane. Mol. Biol. Cell 27, 1101–1119 (2016).

  40. 40.

    Kusumi, A. et al. Membrane mechanisms for signal transduction: the coupling of the meso-scale raft domains to membrane-skeleton-induced compartments and dynamic protein complexes. Semin. Cell Dev. Biol. 23, 126–144 (2012).

  41. 41.

    Kusumi, A., Tsunoyama, T. A., Hirosawa, K. M., Kasai, R. S. & Fujiwara, T. K. Tracking single molecules at work in living cells. Nat. Chem. Biol. 10, 524–532 (2014).

  42. 42.

    Suzuki, K. G. et al. Transient GPI-anchored protein homodimers are units for raft organization and function. Nat. Chem. Biol. 8, 774–783 (2012).

  43. 43.

    Moens, P. D., Digman, M. A. & Gratton, E. Modes of diffusion of cholera toxin bound to GM1 on live cell membrane by image mean square displacement analysis. Biophys. J. 108, 1448–1458 (2015).

  44. 44.

    Groot, R. D. & Warren, P. B. Dissipative particle dynamics: bridging the gap between atomistic and mesoscopic simulation. J. Chem. Phys. 107, 4423–4435 (1997).

  45. 45.

    Williams, P. et al. Quorum sensing and the population-dependent control of virulence. Phil. Trans. R. Soc. Lond. B 355, 667–680 (2000).

  46. 46.

    Davies, D. G. et al. The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280, 295–298 (1998).

  47. 47.

    Schuster, M., Sexton, D. J., Diggle, S. P. & Greenberg, E. P. Acyl-homoserine lactone quorum sensing: from evolution to application. Annu. Rev. Microbiol. 67, 43–63 (2013).

  48. 48.

    Brumatti, G. et al. The caspase-8 inhibitor emricasan combines with the SMAC mimetic birinapant to induce necroptosis and treat acute myeloid leukemia. Sci. Transl Med. 8, 339ra369 (2016).

  49. 49.

    Barreyro, F. J. et al. The pan-caspase inhibitor emricasan (IDN-6556) decreases liver injury and fibrosis in a murine model of non-alcoholic steatohepatitis. Liver Int. 35, 953–966 (2015).

  50. 50.

    Xu, M. et al. Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen. Nat. Med. 22, 1101–1107 (2016).

  51. 51.

    Teplitski, M., Mathesius, U. & Rumbaugh, K. P. Perception and degradation of N-acyl homoserine lactone quorum sensing signals by mammalian and plant cells. Chem. Rev. 111, 100–116 (2011).

  52. 52.

    Miyairi, S. et al. Immunization with 3-oxododecanoyl-l-homoserine lactone–protein conjugate protects mice from lethal Pseudomonas aeruginosa lung infection. J. Med. Microbiol. 55, 1381–1387 (2006).

  53. 53.

    Schwarzer, C. et al. Paraoxonase 2 serves a proapopotic function in mouse and human cells in response to the Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)-homoserine lactone. J. Biol. Chem. 290, 7247–7258 (2015).

  54. 54.

    Horke, S. et al. Novel paraoxonase 2-dependent mechanism mediating the biological effects of the Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxo-dodecanoyl)-l-homoserine lactone. Infect. Immun. 83, 3369–3380 (2015).

  55. 55.

    Tao, S. et al. Paraoxonase 2 modulates a proapoptotic function in LS174T cells in response to quorum sensing molecule N-(3-oxododecanoyl)-l-homoserine lactone. Sci. Rep. 6, 28778 (2016).

  56. 56.

    Wagner, V. E., Bushnell, D., Passador, L., Brooks, A. I. & Iglewski, B. H. Microarray analysis of Pseudomonas aeruginosa quorum-sensing regulons: effects of growth phase and environment. J. Bacteriol. 185, 2080–2095 (2003).

  57. 57.

    Saenz, J. P. et al. Hopanoids as functional analogues of cholesterol in bacterial membranes. Proc. Natl Acad. Sci. USA 112, 11971–11976 (2015).

  58. 58.

    Ritchie, K., Iino, R., Fujiwara, T., Murase, K. & Kusumi, A. The fence and picket structure of the plasma membrane of live cells as revealed by single molecule techniques (review). Mol. Membr. Biol. 20, 13–18 (2003).

  59. 59.

    Barnoud, J., Rossi, G., Marrink, S. J. & Monticelli, L. Hydrophobic compounds reshape membrane domains. PLoS Comput. Biol. 10, e1003873 (2014).

  60. 60.

    Legler, D. F., Doucey, M. A., Cerottini, J. C., Bron, C. & Luescher, I. F. Selective inhibition of CTL activation by a dipalmitoyl-phospholipid that prevents the recruitment of signaling molecules to lipid rafts. FASEB J. 15, 1601–1603 (2001).

  61. 61.

    Joelsson, A. C. & Zhu, J. LacZ-detection of acyl-homoserine lactone quorum-sensing signals. Curr. Protoc. Microbiol. 3, 1C.2.1–1C.2.9 (2006).

  62. 62.

    Clayton, D. A. & Shadel, G. S. Isolation of mitochondria from tissue culture cells. Cold Spring Harb. Protoc. 2014, pdb.prot080002 (2014).

  63. 63.

    Kusumi, A., Sako, Y. & Yamamoto, M. Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys. J. 65, 2021–2040 (1993).

  64. 64.

    Facchini, M., De Fino, I., Riva, C. & Bragonzi, A. Long term chronic Pseudomonas aeruginosa airway infection in mice. J. Vis. Exp. 85, 51019 (2014).

  65. 65.

    Lukinskiene, L. et al. Antimicrobial activity of PLUNC protects against Pseudomonas aeruginosa infection. J. Immunol. 187, 382–390 (2011).

  66. 66.

    Sulahian, T. H., Imrich, A., Deloid, G., Winkler, A. R. & Kobzik, L. Signaling pathways required for macrophage scavenger receptor-mediated phagocytosis: analysis by scanning cytometry. Respir. Res. 9, 59 (2008).

  67. 67.

    de Jong, A. et al. CD1a-autoreactive T cells recognize natural skin oils that function as headless antigens. Nat. Immunol. 15, 177–185 (2014).

  68. 68.

    Tachi, M. & Iwamori, M. Mass spectrometric characterization of cholesterol esters and wax esters in epidermis of fetal, adult and keloidal human skin. Exp. Dermatol. 17, 318–323 (2008).

  69. 69.

    Owen, D. M., Rentero, C., Magenau, A., Abu-Siniyeh, A. & Gaus, K. Quantitative imaging of membrane lipid order in cells and organisms. Nat. Protoc. 7, 24–35 (2011).

  70. 70.

    Jin, L. et al. Characterization and application of a new optical probe for membrane lipid domains. Biophys. J. 90, 2563–2575 (2006).

  71. 71.

    Kim, H. M. et al. A two-photon fluorescent probe for lipid raft imaging: C-laurdan. Chembiochem 8, 553–559 (2007).

  72. 72.

    Collins, M. D. & Gordon, S. E. Giant liposome preparation for imaging and patch-clamp electrophysiology. J. Vis. Exp. 76, 50227 (2013).

  73. 73.

    Pott, T., Bouvrais, H. & Meleard, P. Giant unilamellar vesicle formation under physiologically relevant conditions. Chem. Phys. Lipids 154, 115–119 (2008).

  74. 74.

    Klymchenko, A. S. & Kreder, R. Fluorescent probes for lipid rafts: from model membranes to living cells. Chem. Biol. 21, 97–113 (2014).

  75. 75.

    Groot, R. D. & Warren, P. B. Dissipative particle dynamics: bridging the gap between atomistic and mesoscopic simulation. J. Chem. Phys. 107, 4423–4435 (1997).

  76. 76.

    Tieleman, D. P., Leontiadou, H., Mark, A. E. & Marrink, S. J. Simulation of pore formation in lipid bilayers by mechanical stress and electric fields. J. Am. Chem. Soc. 125, 6382–6383 (2003).

  77. 77.

    Chen, P. et al. Diffusion and directionality of charged nanoparticles on lipid bilayer membrane. ACS Nano 10, 11541–11547 (2016).

  78. 78.

    Vattulainen, I., Karttunen, M., Besold, G. & Polson, J. M. Integration schemes for dissipative particle dynamics simulations: from softly interacting systems towards hybrid models. J. Chem. Phys. 116, 3967–3979 (2002).

  79. 79.

    Smith, K. A., Jasnow, D. & Balazs, A. C. Designing synthetic vesicles that engulf nanoscopic particles. J. Chem. Phys. 127, 084703 (2007).

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We thank P. Greenberg of University of Washington for his expert advice and his gift of P.aeruginosa strains, J. Yuan of Institute of Chemistry of CAS for his assistance on single-molecule imaging data analysis, J. Harrison and H. Almblad for their expert advice on P.aeruginosa mutagenesis and L. Yu of Tsinghua University for DNA constructs. X.F. is supported by the National Natural Science Foundation of China (21735006 and 91413119). Y.S. is supported by the joint Peking-Tsinghua Center for Life Sciences, the National Natural Science Foundation of China General Program (31370878), and by grants from the US NIH (R01AI098995), the Natural Sciences and Engineering Research Council of Canada (RGPIN-355350/396037) and the Canadian Institutes for Health Research (MOP-119295).

Author information


  1. Institute for Immunology, Department of Basic Medical Sciences, Center for Life Sciences, Tsinghua University, Beijing, China

    • Dingka Song
    • , Junchen Meng
    • , Hefei Ruan
    • , Ning Kang
    • , Ying Xu
    • , Xiaobo Wang
    • , Fei Shu
    • , Libing Mu
    • , Tengfei Li
    • , Wenran Ren
    • , Xin Lin
    • , Tie Xia
    •  & Yan Shi
  2. Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, China

    • Jie Cheng
    •  & Junhong Lü
  3. State Key Laboratory of Medicinal Chemical Biology, Key Laboratory of Molecular Microbiology and Technology of the Ministry of Education, Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China

    • Zheng Fan
    •  & Weihui Wu
  4. Key Laboratory of Advanced Materials (MOE), Department of Chemical Engineering, Tsinghua University, Beijing, China

    • Pengyu Chen
    •  & Li-Tang Yan
  5. Department of Microbiology, Immunology and Infectious Diseases, Snyder Institute, University of Calgary, Calgary, Alberta, Canada

    • Zhongyuan Tu
    •  & Yan Shi
  6. Key Laboratory of Molecular Nanostructure and Nanotechnology, Institute of Chemistry, Chinese Academy of Sciences, Beijing, China

    • Nan Li
    •  & Xiaohong Fang
  7. Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    • Jun Zhu
  8. Department of Microbiology, Nanjing Agricultural University, Nanjing, China

    • Jun Zhu
  9. Departments of Cell Biology and Anatomy, Snyder Institute, University of Calgary, Calgary, Alberta, Canada

    • Matthias W. Amrein


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D.S. performed all of the experiments and data analyses unless otherwise specified with assistance from J.M., Z.T., Y.X., X.W., F.S., N.L., W.R. and L.M. N.K. and J.C. performed the AFM analysis. J.L., T.X. and M.W.A. proposed and supervised the AFM experiments. X.L. helped to design the TNFR1 signalling assays. X.F. helped to design the single-molecule imaging experiments. J.Z. helped to design the quorum-sensing mutant experiments. H.R. designed and performed the GUV experiments. T.L. performed the TLC analysis. Z.F. and W.W. designed and constructed the vector-based lasI overexpression version of ΔlasR P.aeruginosa. P.C. and L-T.Y. designed and performed the TNFR1 simulation work. T.X. designed and supervised all imaging work. Y.S. conceptualized the work and wrote the manuscript with assistance from D.S.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Yan Shi.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–6, uncropped gels, Supplementary Tables 1 and 2, Supplementary References, Supplementary Video legends.

  2. Reporting Summary

  3. Supplementary Video 1

    Sample movie of TNFR1 multiple-step fluorescence quenching.

  4. Supplementary Video 2

    Sample movie of AFM scanning of lipid membrane treated with DMSO.

  5. Supplementary Video 3

    Sample movie of AFM scanning of lipid membrane treated with 3oc.

  6. Supplementary Video 4

    Sample movie of TNFR1 single-particle tracking treated with DMSO.

  7. Supplementary Video 5

    Sample movie of TNFR1 single-particle tracking treated with 3oc.

  8. Supplementary Video 6

    Coarse-grained model of TNFR1’s dynamics on plasma membrane.

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