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Exotoxin S secreted by internalized Pseudomonas aeruginosa delays lytic host cell death

Kroken, Abby R., Gajenthra Kumar, Naren, Yahr, Timothy L., Smith, Benjamin E., Nieto, Vincent, Horneman, Hart, Evans, David J., Fleiszig, Suzanne M. J.·2022·PLOS Pathogens
DOI 10.1371/journal.ppat.1010306
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Abstract

The Pseudomonas aeruginosa toxin ExoS, secreted by the type III secretion system (T3SS), supports intracellular persistence via its ADP-ribosyltransferase (ADPr) activity. For epithelial cells, this involves inhibiting vacuole acidification, promoting vacuolar escape, countering autophagy, and niche construction in the cytoplasm and within plasma membrane blebs. Paradoxically, ExoS and other P.aeruginosa T3SS effectors can also have antiphagocytic and cytotoxic activities. Here, we sought to reconcile these apparently contradictory activities of ExoS by studying the relationships between intracellular persistence and host epithelial cell death. Methods involved quantitative imaging and the use of antibiotics that vary in host cell membrane permeability to selectively kill intracellular and extracellular populations after invasion. Results showed that intracellular P.aeruginosa mutants lacking T3SS effector toxins could kill (permeabilize) cells when extracellular bacteria were eliminated. Surprisingly, wild-type strain PAO1 (encoding ExoS, ExoT and ExoY) caused cell death more slowly, the time extended from 5.2 to 9.5 h for corneal epithelial cells and from 10.2 to 13.0 h for HeLa cells. Use of specific mutants/complementation and controls for initial invasion showed that ExoS ADPr activity delayed cell death. Triggering T3SS expression only after bacteria invaded cells using rhamnose-induction in T3SS mutants rescued the ExoS-dependent intracellular phenotype, showing that injected effectors from extracellular bacteria were not required. The ADPr activity of ExoS was further found to support internalization by countering the antiphagocytic activity of both the ExoS and ExoT RhoGAP domains. Together, these results show two additional roles for ExoS ADPr activity in supporting the intracellular lifestyle of P.aeruginosa; suppression of host cell death to preserve a replicative niche and inhibition of T3SS effector antiphagocytic activities to allow invasion. These findings add to the growing body of evidence that ExoS-encoding (invasive)P.aeruginosa strains can be facultative intracellular pathogens, and that intracellularly secreted T3SS effectors contribute to pathogenesis.

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Research works printed but not linked: 73

  1. D Church, 2006, Burn wound infections, Clin Microbiol Rev, 19, 403, 10.1128/CMR.19.2.403-434.2006 · DOI · Paper page
  2. JB Lyczak, 2000, Establishment of Pseudomonas aeruginosa infection: lessons from a versatile opportunist, Microbes Infect, 2, 1051, 10.1016/S1286-4579(00)01259-4 · DOI · Paper page
  3. LE Nicolle, 2005, Ammi Canada Guidelines Committee. Complicated urinary tract infection in adults, Can J Infect Dis Med Microbiol, 16, 349, 10.1155/2005/385768 · DOI · Paper page
  4. JR Cope, 2015, Contact Lens Wearer Demographics and Risk Behaviors for Contact Lens-Related Eye Infections—United States, 2014, MMWR Morb Mortal Wkly Rep, 64, 865, 10.15585/mmwr.mm6432a2 · doi:10.15585/mmwr.mm6432a2
  5. TL Yahr, 1996, Exoenzyme S of Pseudomonas aeruginosa is secreted by a type III pathway, Mol Microbiol, 22, 991, 10.1046/j.1365-2958.1996.01554.x · DOI · Paper page
  6. M Galle, 2012, Structure and function of the Type III secretion system of Pseudomonas aeruginosa, Curr Protein Pept Sci, 13, 831, 10.2174/138920312804871210 · DOI · Paper page
  7. SM Fleiszig, 1997, Pseudomonas aeruginosa-mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S, Infect Immun, 65, 579, 10.1128/iai.65.2.579-586.1997 · DOI · Paper page
  8. H Silistre, 2021, Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells, Front Microbiol, 12, 666097, 10.3389/fmicb.2021.666097 · doi:10.3389/fmicb.2021.666097
  9. V Hritonenko, 2011, Adenylate cyclase activity of Pseudomonas aeruginosa ExoY can mediate bleb-niche formation in epithelial cells and contributes to virulence, Microb Pathog, 51, 305, 10.1016/j.micpath.2011.08.001 · doi:10.1016/j.micpath.2011.08.001
  10. TL Yahr, 1998, ExoY, an adenylate cyclase secreted by the Pseudomonas aeruginosa type III system, Proc Natl Acad Sci U S A, 95, 13899, 10.1073/pnas.95.23.13899 · DOI · Paper page
  11. J Jeon, 2017, T3SS effector ExoY reduces inflammasome-related responses by suppressing bacterial motility and delaying activation of NF-kappaB and caspase-1, FEBS J, 284, 3392, 10.1111/febs.14199 · DOI
  12. TC Stevens, 2014, The Pseudomonas aeruginosa exoenzyme Y impairs endothelial cell proliferation and vascular repair following lung injury, Am J Physiol Lung Cell Mol Physiol, 306, L915, 10.1152/ajplung.00135.2013 · DOI
  13. BA Cowell, 2005, Actin cytoskeleton disruption by ExoY and its effects on Pseudomonas aeruginosa invasion, FEMS Microbiol Lett, 250, 71, 10.1016/j.femsle.2005.06.044 · DOI
  14. H Sato, 2004, ExoU is a potent intracellular phospholipase, Mol Microbiol, 53, 1279, 10.1111/j.1365-2958.2004.04194.x · DOI · Paper page
  15. V Finck-Barbancon, 1997, ExoU expression by Pseudomonas aeruginosa correlates with acute cytotoxicity and epithelial injury, Mol Microbiol, 25, 547, 10.1046/j.1365-2958.1997.4891851.x · DOI · Paper page
  16. SM Fleiszig, 1996, Relationship between cytotoxicity and corneal epithelial cell invasion by clinical isolates of Pseudomonas aeruginosa, Infect Immun, 64, 2288, 10.1128/iai.64.6.2288-2294.1996 · DOI · Paper page
  17. H Feltman, 2001, Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa, Microbiology (Reading)., 147, 2659, 10.1099/00221287-147-10-2659 · DOI · Paper page
  18. JT Barbieri, 2004, Pseudomonas aeruginosa ExoS and ExoT, Rev Physiol Biochem Pharmacol, 152, 79, 10.1007/s10254-004-0031-7 · DOI · Paper page
  19. BA Cowell, 2000, ExoT of cytotoxic Pseudomonas aeruginosa prevents uptake by corneal epithelial cells, Infect Immun, 68, 403, 10.1128/IAI.68.1.403-406.2000 · doi:10.1128/iai.68.1.403-406.2000
  20. L Garrity-Ryan, 2000, The arginine finger domain of ExoT contributes to actin cytoskeleton disruption and inhibition of internalization of Pseudomonas aeruginosa by epithelial cells and macrophages, Infect Immun, 68, 7100, 10.1128/IAI.68.12.7100-7113.2000 · DOI · Paper page
  21. UM Goehring, 1999, The N-terminal domain of Pseudomonas aeruginosa exoenzyme S is a GTPase-activating protein for Rho GTPases, J Biol Chem, 274, 36369, 10.1074/jbc.274.51.36369 · DOI · Paper page
  22. J Sun, 2003, Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 regulator of kinase (Crk) proteins, J Biol Chem, 278, 32794, 10.1074/jbc.M304290200 · DOI
  23. Q Deng, 2008, Modulation of host cell endocytosis by the type III cytotoxin, Pseudomonas ExoS, Traffic, 9, 1948, 10.1111/j.1600-0854.2008.00808.x · DOI
  24. MJ Riese, 2001, ADP ribosylation of Arg41 of Rap by ExoS inhibits the ability of Rap to interact with its guanine nucleotide exchange factor, C3G. Biochemistry, 40, 3289, 10.1021/bi002729q · doi:10.1021/bi002729q
  25. AK Ganesan, 1999, Pseudomonas aeruginosa exoenzyme S disrupts Ras-mediated signal transduction by inhibiting guanine nucleotide exchange factor-catalyzed nucleotide exchange, J Biol Chem, 274, 21823, 10.1074/jbc.274.31.21823 · DOI
  26. AW Maresso, 2007, Pseudomonas aeruginosa ExoS ADP-ribosyltransferase inhibits ERM phosphorylation, Cell Microbiol, 9, 97, 10.1111/j.1462-5822.2006.00770.x · doi:10.1111/j.1462-5822.2006.00770.x
  27. J Coburn, 1989, Exoenzyme S of Pseudomonas aeruginosa ADP-ribosylates the intermediate filament protein vimentin, Infect Immun, 57, 996, 10.1128/iai.57.3.996-998.1989 · doi:10.1128/iai.57.3.996-998.1989
  28. AA DiNovo, 2006, ADP-ribosylation of cyclophilin A by Pseudomonas aeruginosa exoenzyme S, Biochemistry, 45, 4664, 10.1021/bi0513554 · doi:10.1021/bi0513554
  29. EM McGuffie, 1998, Modification of Ras in eukaryotic cells by Pseudomonas aeruginosa exoenzyme S, Infect Immun, 66, 2607, 10.1128/IAI.66.6.2607-2613.1998 · DOI
  30. EM McGuffie, 1999, Differential sensitivity of human epithelial cells to Pseudomonas aeruginosa exoenzyme S, Infect Immun, 67, 3494, 10.1128/IAI.67.7.3494-3503.1999 · doi:10.1128/iai.67.7.3494-3503.1999
  31. JE Fraylick, 2002, Eukaryotic cell determination of ExoS ADP-ribosyltransferase substrate specificity, Biochem Biophys Res Commun, 291, 91, 10.1006/bbrc.2002.6402 · doi:10.1006/bbrc.2002.6402
  32. EA Rucks, 2003, Cell line differences in bacterially translocated ExoS ADP-ribosyltransferase substrate specificity, Microbiology (Reading)., 149, 319, 10.1099/mic.0.25985-0 · DOI
  33. MJ Riese, 2002, Auto-ADP-ribosylation of Pseudomonas aeruginosa ExoS, J Biol Chem, 277, 12082, 10.1074/jbc.M109039200 · DOI
  34. SM Fleiszig, 1995, Pseudomonas aeruginosa invasion of and multiplication within corneal epithelial cells in vitro, Infect Immun, 63, 4072, 10.1128/iai.63.10.4072-4077.1995 · doi:10.1128/iai.63.10.4072-4077.1995
  35. AA Angus, 2008, Pseudomonas aeruginosa induces membrane blebs in epithelial cells, which are utilized as a niche for intracellular replication and motility, Infect Immun, 76, 1992, 10.1128/IAI.01221-07 · doi:10.1128/iai.01221-07
  36. AL Jolly, 2015, Pseudomonas aeruginosa-induced bleb-niche formation in epithelial cells is independent of actinomyosin contraction and enhanced by loss of cystic fibrosis transmembrane-conductance regulator osmoregulatory function, mBio, 6, e02533, 10.1128/mBio.02533-14 · doi:10.1128/mbio.02533-14
  37. C Penaranda, 2021, Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection., PLoS Pathog, 17, e1009534, 10.1371/journal.ppat.1009534 · DOI
  38. SM Fleiszig, 1994, Pseudomonas aeruginosa invades corneal epithelial cells during experimental infection, Infect Immun, 62, 3485, 10.1128/iai.62.8.3485-3493.1994 · doi:10.1128/iai.62.8.3485-3493.1994
  39. DJ Evans, 1998, Pseudomonas aeruginosa invasion and cytotoxicity are independent events, both of which involve protein tyrosine kinase activity, Infect Immun, 66, 1453, 10.1128/IAI.66.4.1453-1459.1998 · DOI
  40. AA Angus, 2010, The ADP-ribosylation domain of Pseudomonas aeruginosa ExoS is required for membrane bleb niche formation and bacterial survival within epithelial cells, Infect Immun, 78, 4500, 10.1128/IAI.00417-10 · DOI
  41. C Vareechon, 2017, Pseudomonas aeruginosa Effector ExoS Inhibits ROS Production in Human Neutrophils., Cell Host Microbe, 21, 611, 10.1016/j.chom.2017.04.001 · DOI
  42. AR Kroken, 2018, The Impact of ExoS on Pseudomonas aeruginosa Internalization by Epithelial Cells Is Independent of fleQ and Correlates with Bistability of Type Three Secretion System Gene Expression, mBio, 9, 10.1128/mBio.00668-18 · doi:10.1128/mbio.00668-18
  43. SR Heimer, 2013, Pseudomonas aeruginosa utilizes the type III secreted toxin ExoS to avoid acidified compartments within epithelial cells, PLoS One, 8, e73111, 10.1371/journal.pone.0073111 · DOI
  44. L Rao, 2021, Pseudomonas aeruginosa survives in epithelia by ExoS-mediated inhibition of autophagy and mTOR, EMBO Rep, 22, e50613, 10.15252/embr.202050613 · DOI
  45. V Nieto, 2019, Type IV Pili Can Mediate Bacterial Motility within Epithelial Cells, mBio, 10, 10.1128/mBio.02880-18 · DOI
  46. ML Urbanowski, 2007, Translocation of ExsE into Chinese hamster ovary cells is required for transcriptional induction of the Pseudomonas aeruginosa type III secretion system, Infect Immun, 75, 4432, 10.1128/IAI.00664-07 · doi:10.1128/iai.00664-07
  47. D Ince, 2015, Secretion of Flagellar Proteins by the Pseudomonas aeruginosa Type III Secretion-Injectisome System, J Bacteriol, 197, 2003, 10.1128/JB.00030-15 · DOI
  48. RE Vance, 2005, Role of the type III secreted exoenzymes S, T, and Y in systemic spread of Pseudomonas aeruginosa PAO1 in vivo, Infect Immun, 73, 1706, 10.1128/IAI.73.3.1706-1713.2005 · DOI · Paper page
  49. TL Yahr, 1997, Identification of type III secreted products of the Pseudomonas aeruginosa exoenzyme S regulon, J Bacteriol, 179, 7165, 10.1128/jb.179.22.7165-7168.1997 · DOI · Paper page
  50. KJ Pederson, 2000, Intracellular localization and processing of Pseudomonas aeruginosa ExoS in eukaryotic cells, Mol Microbiol, 37, 287, 10.1046/j.1365-2958.2000.01990.x · DOI
  51. KJ Pederson, 2002, Intracellular localization modulates targeting of ExoS, a type III cytotoxin, to eukaryotic signalling proteins, Mol Microbiol, 46, 1381, 10.1046/j.1365-2958.2002.03256.x · doi:10.1046/j.1365-2958.2002.03256.x
  52. J. Engel, Pseudomonas aeruginosa Internalization by Non-Phagocytic Cells., Pseudomonas. 52007, 343
  53. TG Sana, 2015, Internalization of Pseudomonas aeruginosa Strain PAO1 into Epithelial Cells Is Promoted by Interaction of a T6SS Effector with the Microtubule Network, mBio, 6, e00712
  54. M Cisz, 2008, ExoS controls the cell contact-mediated switch to effector secretion in Pseudomonas aeruginosa, J Bacteriol, 190, 2726, 10.1128/JB.01553-07 · DOI
  55. AM McDermott, 2013, Antimicrobial compounds in tears, Exp Eye Res, 117, 53, 10.1016/j.exer.2013.07.014 · DOI · Paper page
  56. AT Gewirtz, 2001, Cutting edge: bacterial flagellin activates basolaterally expressed TLR5 to induce epithelial proinflammatory gene expression, J Immunol, 167, 1882, 10.4049/jimmunol.167.4.1882 · DOI · Paper page
  57. DH Wyllie, 2000, Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses, J Immunol, 165, 7125, 10.4049/jimmunol.165.12.7125 · DOI · Paper page
  58. T Grandjean, 2017, The human NAIP-NLRC4-inflammasome senses the Pseudomonas aeruginosa T3SS inner-rod protein, Int Immunol, 29, 377, 10.1093/intimm/dxx047 · DOI
  59. EA Miao, 2010, Innate immune detection of the type III secretion apparatus through the NLRC4 inflammasome, Proc Natl Acad Sci U S A, 107, 3076, 10.1073/pnas.0913087107 · DOI · Paper page
  60. P Garai, 2019, Killing from the inside: Intracellular role of T3SS in the fate of Pseudomonas aeruginosa within macrophages revealed by mgtC and oprF mutants, PLoS Pathog, 15, e1007812, 10.1371/journal.ppat.1007812 · DOI
  61. LH Travassos, 2005, Nod1 participates in the innate immune response to Pseudomonas aeruginosa, J Biol Chem, 280, 36714, 10.1074/jbc.M501649200 · DOI · Paper page
  62. SE Girardin, 2003, Nod2 is a general sensor of peptidoglycan through muramyl dipeptide (MDP) detection., J Biol Chem, 278, 8869, 10.1074/jbc.C200651200 · DOI · Paper page
  63. J Kortmann, 2015, Cutting Edge: Inflammasome Activation in Primary Human Macrophages Is Dependent on Flagellin, J Immunol, 195, 815, 10.4049/jimmunol.1403100 · DOI · Paper page
  64. N Kayagaki, 2013, Noncanonical inflammasome activation by intracellular LPS independent of TLR4, Science, 341, 1246, 10.1126/science.1240248 · DOI · Paper page
  65. M Galle, 2008, The Pseudomonas aeruginosa Type III secretion system plays a dual role in the regulation of caspase-1 mediated IL-1beta maturation, J Cell Mol Med, 12, 1767, 10.1111/j.1582-4934.2007.00190.x · DOI
  66. J Jia, 2006, Expression of Pseudomonas aeruginosa toxin ExoS effectively induces apoptosis in host cells, Infect Immun, 74, 6557, 10.1128/IAI.00591-06 · DOI
  67. Y Sun, 2012, ExoS and ExoT ADP ribosyltransferase activities mediate Pseudomonas aeruginosa keratitis by promoting neutrophil apoptosis and bacterial survival, J Immunol, 188, 1884, 10.4049/jimmunol.1102148 · DOI
  68. KH Choi, 2006, mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa., Nat Protoc, 1, 153, 10.1038/nprot.2006.24 · DOI · Paper page
  69. DM Robertson, 2005, Characterization of growth and differentiation in a telomerase-immortalized human corneal epithelial cell line, Invest Ophthalmol Vis Sci, 46, 470, 10.1167/iovs.04-0528 · DOI · Paper page
  70. SM Fleiszig, 1997, Epithelial cell polarity affects susceptibility to Pseudomonas aeruginosa invasion and cytotoxicity, Infect Immun, 65, 2861, 10.1128/iai.65.7.2861-2867.1997 · DOI · Paper page
  71. J Schindelin, 2012, Fiji: an open-source platform for biological-image analysis, Nat Methods, 9, 676, 10.1038/nmeth.2019 · DOI · Paper page
  72. D Legland, 2016, MorphoLibJ: integrated library and plugins for mathematical morphology with ImageJ, Bioinformatics, 32, 3532, 10.1093/bioinformatics/btw413 · DOI · Paper page
  73. W. McKinney, 2010, Data Structures for Statistical Computing in Python, Proceedings of the 9th Python in Science Conference, 56, 10.25080/Majora-92bf1922-00a · DOI · Paper page

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Metadata

Authors
Kroken, A. R., Gajenthra Kumar, N., Yahr, T. L., Smith, B. E., Nieto, V., Horneman, H., Evans, D. J., & Fleiszig, S. M. J.
ORCID
Fleiszig, Suzanne M. J.
Venue
PLOS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2022-02-07
Volume
18
Issue
2
Pages
e1010306
ISSN
1553-7374
DOI
10.1371/journal.ppat.1010306
Cited by
45
References
0
Language
en
Update notices
new_version (2022-02-17)
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
National Eye Institute (EY011221, EY025969)

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Kroken, A. R., Gajenthra Kumar, N., Yahr, T. L., Smith, B. E., Nieto, V., Horneman, H., Evans, D. J., & Fleiszig, S. M. J. (2022). Exotoxin S secreted by internalized Pseudomonas aeruginosa delays lytic host cell death. PLOS Pathogens. https://doi.org/10.1371/journal.ppat.1010306

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