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Novel roles for two-component regulatory systems in cytotoxicity and virulence-related properties in Pseudomonas aeruginosa

  • Received: 14 December 2017 Accepted: 26 February 2018 Published: 08 March 2018
  • The rapid adaptation of the opportunistic bacterial pathogen Pseudomonas aeruginosa to various growth modes and environmental conditions is controlled in part through diverse two-component regulatory systems. Some of these systems are well studied, but the majority are poorly characterized, even though it is likely that several of these systems contribute to virulence. Here, we screened all available strain PA14 mutants in 50 sensor kinases, 50 response regulators and 5 hybrid sensor/regulators, for contributions to cytotoxicity against cultured human bronchial epithelial cells, as assessed by the release of cytosolic lactate dehydrogenase. This enabled the identification of 8 response regulators and 3 sensor kinases that caused substantial decreases in cytotoxicity, and 5 response regulators and 8 sensor kinases that significantly increased cytotoxicity by 15–58% or more. These regulators were additionally involved in motility, adherence, type 3 secretion, production of cytotoxins, and the development of biofilms. Here we investigated in more detail the roles of FleSR, PilSR and WspR. Not all cognate pairs contributed to cytotoxicity (e.g. PhoPQ, PilSR) in the same way and some differences could be detected between the same mutants in PAO1 and PA14 strain backgrounds (e.g. FleSR, PhoPQ). This study highlights the potential importance of these regulators and their downstream targets on pathogenesis and demonstrates that cytotoxicity can be regulated by several systems and that their contributions are partly dependent on strain background.

    Citation: Shaan L. Gellatly, Manjeet Bains, Elena B.M. Breidenstein, Janine Strehmel, Fany Reffuveille, Patrick K. Taylor, Amy T.Y. Yeung, Joerg Overhage, Robert E.W. Hancock. Novel roles for two-component regulatory systems in cytotoxicity and virulence-related properties in Pseudomonas aeruginosa[J]. AIMS Microbiology, 2018, 4(1): 173-191. doi: 10.3934/microbiol.2018.1.173

    Related Papers:

  • The rapid adaptation of the opportunistic bacterial pathogen Pseudomonas aeruginosa to various growth modes and environmental conditions is controlled in part through diverse two-component regulatory systems. Some of these systems are well studied, but the majority are poorly characterized, even though it is likely that several of these systems contribute to virulence. Here, we screened all available strain PA14 mutants in 50 sensor kinases, 50 response regulators and 5 hybrid sensor/regulators, for contributions to cytotoxicity against cultured human bronchial epithelial cells, as assessed by the release of cytosolic lactate dehydrogenase. This enabled the identification of 8 response regulators and 3 sensor kinases that caused substantial decreases in cytotoxicity, and 5 response regulators and 8 sensor kinases that significantly increased cytotoxicity by 15–58% or more. These regulators were additionally involved in motility, adherence, type 3 secretion, production of cytotoxins, and the development of biofilms. Here we investigated in more detail the roles of FleSR, PilSR and WspR. Not all cognate pairs contributed to cytotoxicity (e.g. PhoPQ, PilSR) in the same way and some differences could be detected between the same mutants in PAO1 and PA14 strain backgrounds (e.g. FleSR, PhoPQ). This study highlights the potential importance of these regulators and their downstream targets on pathogenesis and demonstrates that cytotoxicity can be regulated by several systems and that their contributions are partly dependent on strain background.


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    [1] Gellatly SL, Hancock REW (2013) Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathog Dis 67: 159–173. doi: 10.1111/2049-632X.12033
    [2] Schurek KN, Breidenstein EBM, Hancock REW (2012) Pseudomonas aeruginosa: a persistent pathogen in cystic fibrosis and hospital-associated infections, In: Dougherty TJ, Pucci MJ, Editors, Antibiotic drug discovery and Development, Springer Publishing Company.
    [3] Roy-Burman A, Savel RH, Racine S, et al. (2001) Type III protein secretion is associated with death in lower respiratory and systemic Pseudomonas aeruginosa infections. J Infect Dis 183: 1767–1774. doi: 10.1086/320737
    [4] Hauser AR, Cobb E, Bodi M, et al. (2002) Type III protein secretion is associated with poor clinical outcomes in patients with ventilator-associated pneumonia caused by Pseudomonas aeruginosa. Crit Care Med 30: 521–528. doi: 10.1097/00003246-200203000-00005
    [5] Mikkelsen H, McMullan R, Filloux A (2011) The Pseudomonas aeruginosa reference strain PA14 displays increased virulence due to a mutation in ladS. PLoS One 6: e29113. doi: 10.1371/journal.pone.0029113
    [6] Ramirez JC, Fleiszig SM, Sullivan AB, et al. (2012) Traversal of multilayered corneal epithelia by cytotoxic Pseudomonas aeruginosa requires the phospholipase domain of ExoU. Invest Ophthalmol Vis Sci 53: 448–453. doi: 10.1167/iovs.11-8999
    [7] Gooderham WJ, Hancock REW (2009) Regulation of virulence and antibiotic resistance by two-component regulatory systems in Pseudomonas aeruginosa. FEMS Microbiol Rev 33: 279–294. doi: 10.1111/j.1574-6976.2008.00135.x
    [8] Yeung AT, Bains M, Hancock RE (2011) The sensor kinase CbrA is a global regulator that modulates metabolism, virulence, and antibiotic resistance in Pseudomonas aeruginosa. J Bacteriol 193: 918–931. doi: 10.1128/JB.00911-10
    [9] Gooderham WJ, Gellatly SL, Sanschagrin F, et al. (2009) The sensor kinase PhoQ mediates virulence in Pseudomonas aeruginosa. Microbiology 155: 699–711. doi: 10.1099/mic.0.024554-0
    [10] Liberati NT, Urbach JM, Miyata S, et al. (2006) An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants. Proc Natl Acad Sci USA 103: 2833–2838. doi: 10.1073/pnas.0511100103
    [11] Lewenza S, Falsafi RK, Winsor G, et al. (2005) Construction of a mini-Tn5-luxCDABE mutant library in Pseudomonas aeruginosa PAO1: A tool for identifying differentially regulated genes. Genome Res 15: 583–589. doi: 10.1101/gr.3513905
    [12] Jacobs MA, Alwood A, Thaipisuttikul I, et al. (2003) Comprehensive transposon mutant library of Pseudomonas aeruginosa. Proc Natl Acad Sci USA 100: 14339–14344. doi: 10.1073/pnas.2036282100
    [13] Macfarlane ELA, Kwasnicka A, Ochs MM, et al. (1999) PhoP-PhoQ homologues in Pseudomonas aeruginosa regulate expression of the outer-membrane protein OprH and polymyxin B resistance. Mol Microbiol 34: 305–316. doi: 10.1046/j.1365-2958.1999.01600.x
    [14] Breidenstein EB, Khaira BK, Wiegand I, et al. (2008) Complex ciprofloxacin resistome revealed by screening a Pseudomonas aeruginosa mutant library for altered susceptibility. Antimicrob Agents Ch 52: 4486–4491. doi: 10.1128/AAC.00222-08
    [15] Choi KH, Gaynor JB, White KG, et al. (2005) A Tn7-based broad-range bacterial cloning and expression system. Nat Methods 2: 443–448. doi: 10.1038/nmeth765
    [16] Choi KH, Schweizer HP (2006) Mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Nat Protoc 1: 153–161. doi: 10.1038/nprot.2006.24
    [17] Gruenert DC, Basbaum CB, Welsh MJ, et al. (1988) Characterization of human tracheal epithelial cells transformed by an origin-defective simian virus 40. Proc Natl Acad Sci USA 85: 5951–5955. doi: 10.1073/pnas.85.16.5951
    [18] Yeung AT, Torfs EC, Jamshidi F, et al. (2009) Swarming of Pseudomonas aeruginosa is controlled by a broad spectrum of transcriptional regulators, including MetR. J Bacteriol 191: 5592–5602. doi: 10.1128/JB.00157-09
    [19] Overhage J, Lewenza S, Marr AK, et al. (2007) Identification of genes involved in swarming motility using a Pseudomonas aeruginosa PAO1 mini-Tn5-lux mutant library. J Bacteriol 189: 2164–2169. doi: 10.1128/JB.01623-06
    [20] O'Toole GA, Kolter R (1998) Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol 30: 295–304. doi: 10.1046/j.1365-2958.1998.01062.x
    [21] Gellatly SL, Needham B, Madera L, et al. (2012) The Pseudomonas aeruginosa PhoP-PhoQ two-component regulatory system is induced upon interaction with epithelial cells and controls cytotoxicity and inflammation. Infect Immun 80: 3122–3131. doi: 10.1128/IAI.00382-12
    [22] Wu W, Badrane H, Arora S, et al. (2004) MucA-mediated coordination of type III secretion and alginate synthesis in Pseudomonas aeruginosa. J Bacteriol 186: 7575–7585. doi: 10.1128/JB.186.22.7575-7585.2004
    [23] Intile PJ, Diaz MR, Urbanowski ML, et al. (2014) The AlgZR two-component system recalibrates the RsmAYZ posttranscriptional regulatory system to inhibit expression of the Pseudomonas aeruginosa type III secretion system. J Bacteriol 196: 357–366. doi: 10.1128/JB.01199-13
    [24] Carterson AJ, Morici LA, Jackson DW, et al. (2004) The transcriptional regulator AlgR controls cyanide production in Pseudomonas aeruginosa. J Bacteriol 186: 6837–6844. doi: 10.1128/JB.186.20.6837-6844.2004
    [25] Whitchurch CB, Alm RA, Mattick JS (1996) The alginate regulator AlgR and an associated sensor FimS are required for twitching motility in Pseudomonas aeruginosa. Proc Natl Acad Sci USA 93: 9839–9843. doi: 10.1073/pnas.93.18.9839
    [26] Lizewski SE, Lundberg DS, Schurr MJ (2002) The transcriptional regulator AlgR is essential for Pseudomonas aeruginosa pathogenesis. Infect Immun 70: 6083–6093. doi: 10.1128/IAI.70.11.6083-6093.2002
    [27] Jiang J, Gu BH, Albright LM, et al. (1989) Conservation between coding and regulatory elements of Rhizobium meliloti and Rhizobium leguminosarum dct genes. J Bacteriol 171: 5244–5253. doi: 10.1128/jb.171.10.5244-5253.1989
    [28] Wang YP, Birkenhead K, Boesten B, et al. (1989) Genetic analysis and regulation of the Rhizobium meliloti genes controlling C4-dicarboxylic acid transport. Gene 85: 135–144. doi: 10.1016/0378-1119(89)90473-3
    [29] Valentini M, Storelli N, Lapouge K (2011) Identification of C4-dicarboxylate transport systems in Pseudomonas aerguinosa PAO1. J Bacteriol 193: 4307–4316. doi: 10.1128/JB.05074-11
    [30] Ritchings BW, Almira EC, Lory S, et al. (1995) Cloning and phenotypic characterization of fleS and fleR, new response regulators of Pseudomonas aeruginosa which regulate motility and adhesion to mucin. Infect Immun 63: 4868–4876.
    [31] Duan Q, Zhou M, Zhu L, et al. (2013) Flagella and bacterial pathogenicity. J Basic Microb 53: 1–8. doi: 10.1002/jobm.201100335
    [32] Haiko J, Westerlund-Wikström B (2013) The role of the bacterial flagellum in adhesion and virulence. Biology 2: 1242–1267. doi: 10.3390/biology2041242
    [33] Kohler T, Curty LK, Barja F, et al. (2000) Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J Bacteriol 182: 5990–5996. doi: 10.1128/JB.182.21.5990-5996.2000
    [34] Dasgupta N, Wolfgang MC, Goodman AL, et al. (2003) A four-tiered transcriptional regulatory circuit controls flagellar biogenesis in Pseudomonas aeruginosa. Mol Microbiol 50: 809–824. doi: 10.1046/j.1365-2958.2003.03740.x
    [35] Arora SK, Ritchings BW, Almira EC, et al. (1997) A transcriptional activator, FleQ, regulates mucin adhesion and flagellar gene expression in Pseudomonas aeruginosa in a cascade manner. J Bacteriol 179: 5574–5581. doi: 10.1128/jb.179.17.5574-5581.1997
    [36] Mattick JS (2002) Type IV pili and twitching motility. Annu Rev Microbiol 56: 289–314. doi: 10.1146/annurev.micro.56.012302.160938
    [37] Kelly NM, Kluftinger JL, Pasloske BL, et al. (1989) Pseudomonas aeruginosa pili as ligands for nonopsonic phagocytosis by fibronectin-stimulated macrophages. Infect Immun 57: 3841–3845.
    [38] Chiang P, Burrows LL (2003) Biofilm formation by hyperpiliated mutants of Pseudomonas aeruginosa. J Bacteriol 185: 2374–2378. doi: 10.1128/JB.185.7.2374-2378.2003
    [39] Vallet I, Olson JW, Lory S, et al. (2001) The chaperone/usher pathways of Pseudomonas aeruginosa: identification of fimbrial gene clusters (cup) and their involvement in biofilm formation. Proc Natl Acad Sci USA 98: 6911–6916. doi: 10.1073/pnas.111551898
    [40] Harmsen M, Yang L, Pamp SJ, et al. (2010) An update on Pseudomonas aeruginosa biofilm formation, tolerance, and dispersal. FEMS Immunol Med Microbiol 59: 253–268. doi: 10.1111/j.1574-695X.2010.00690.x
    [41] Craig L, Pique ME, Tainer JA (2004) Type IV pilus structure and bacterial pathogenicity. Nat Rev Microbiol 2: 363–378. doi: 10.1038/nrmicro885
    [42] D'Argenio DA, Calfee MW, Rainey PB, et al. (2002) Autolysis and autoaggregation in Pseudomonas aeruginosa colony morphology mutants. J Bacteriol 184: 6481–6489. doi: 10.1128/JB.184.23.6481-6489.2002
    [43] Jenal U, Malone J (2006) Mechanisms of cyclic-di-GMP signaling in bacteria. Annu Rev Genet 40: 385–407. doi: 10.1146/annurev.genet.40.110405.090423
    [44] Giraud C, Bernard CS, Calderon V, et al. (2011) The PprA-PprB two-component system activates CupE, the first non-archetypal Pseudomonas aeruginosa chaperone-usher pathway system assembling fimbriae. Environ Microbiol 13: 666–683. doi: 10.1111/j.1462-2920.2010.02372.x
    [45] Sato A, Iwasaki A (2005) Peyer's patch dendritic cells as regulators of mucosal adaptive immunity. Cell Mol Life Sci 62: 1333–1338. doi: 10.1007/s00018-005-5037-z
    [46] Wu X, Wang H, Zhao X (2008) Antimicrobial studies with the Pseudomonas aeruginosa two-allele library require caution. Antimicrob Agents Ch 52: 3826–3827. doi: 10.1128/AAC.00419-08
    [47] Held K, Ramage E, Jacobs M, et al. (2012) Sequence-verified two-allele transposon mutant library for Pseudomonas aeruginosa PAO1. J Bacteriol 194: 6387–6389. doi: 10.1128/JB.01479-12
    [48] Macfarlane EL, Kwasnicka A, Hancock RE (2000) Role of Pseudomonas aeruginosa PhoP-PhoQ in resistance to antimicrobial cationic peptides and aminoglycosides. Microbiology 146: 2543–2554. doi: 10.1099/00221287-146-10-2543
    [49] Yeung AT, Janot L, Pena OM, et al. (2014) Requirement of the Pseudomonas aeruginosa CbrA sensor kinase for full virulence in a murine acute lung infection model. Infect Immun 82: 1256–1267. doi: 10.1128/IAI.01527-13
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