1887

Journal of Medical Microbiology logo

Volume 69, Issue 3

Investigation of LuxS-mediated quorum sensing in Open Access

Abstract

Autoinducer-2 (AI-2) quorum sensing is a bacterial communication system that responds to cell density. The system requires activity to produce AI-2, which can regulate gene expression and processes such as biofilm formation.

To investigate the role of in biofilm formation and gene expression in the nosocomial pathogen .

A gene deletion was made in KP563, an extensively drug-resistant isolate. AI-2 production was assessed in wild-type and strains grown in media supplemented with different carbohydrates. Potential roles of in biofilm formation were investigated using a microtiter plate biofilm assay and scanning electron microscopy. Quantitative RT-PCR evaluated the expression of lipopolysaccharide ( and ), polysaccharide (), and type 3 fimbriae () synthesis genes in wild-type and mutant biofilm extracts.

AI-2 production was dependent on the presence of . AI-2 accumulation was highest during early stationary phase in media supplemented with glucose, sucrose or glycerol. Changes in biofilm architecture were observed in the mutant, with less surface coverage and reduced macrocolony formation; however, no differences in biofilm formation between the wild-type and mutant using a microtiter plate assay were observed. In mutant biofilm extracts, the expression of was down-regulated, and the expression of , which encodes a porin for poly-β−1,6-N-acetyl--glucosamine (PNAG) polysaccharide secretion, was upregulated.

Relationships among AI-2-mediated quorum sensing, biofilm formation and gene expression of outer-membrane components were identified in . These inter-connected processes could be important for bacterial group behaviour and persistence.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): biofilm , Klebsiella pneumoniae , luxS and quorum sensing
Funding
This study was supported by the:
  • National Natural Science Foundation of China (Award no.81741059)
    • Principle Award Recipient: Tieli Zhou
  • Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents (Award no. [2012]241)
    • Principle Award Recipient: Tieli Zhou
  • National Health and Medical Research Council (Award 1092262)
    • Principle Award Recipient: Trevor Lithgow
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001148
2020-01-28
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jmm/69/3/402.html?itemId=/content/journal/jmm/10.1099/jmm.0.001148&mimeType=html&fmt=ahah

References

  1. Laganenka L, Colin R, Sourjik V. Chemotaxis towards autoinducer 2 mediates autoaggregation in Escherichia coli . Nat Commun 2016; 7:12984 [View Article]
     [Google Scholar]
  2. Vendeville A, Winzer K, Heurlier K, Tang CM, Hardie KR. Making 'sense' of metabolism: autoinducer-2, LuxS and pathogenic bacteria. Nat Rev Microbiol 2005; 3:383–396 [View Article]
     [Google Scholar]
  3. Li Z, Nair SK. Quorum sensing: how bacteria can coordinate activity and synchronize their response to external signals?. Protein Sci 2012; 21:1403–1417 [View Article]
     [Google Scholar]
  4. Pereira CS, Thompson JA, Xavier KB. AI-2-mediated signalling in bacteria. FEMS Microbiol Rev 2013; 37:156–181 [View Article]
     [Google Scholar]
  5. Balestrino D, Haagensen JAJ, Rich C, Forestier C. Characterization of type 2 quorum sensing in Klebsiella pneumoniae and relationship with biofilm formation. J Bacteriol 2005; 187:2870–2880 [View Article]
     [Google Scholar]
  6. Herzberg M, Kaye IK, Peti W, Wood TK, YdgG WTK. YdgG (TqsA) controls biofilm formation in Escherichia coli K-12 through autoinducer 2 transport. J Bacteriol 2006; 188:587–598 [View Article]
     [Google Scholar]
  7. Xavier KB, Bassler BL. Regulation of uptake and processing of the quorum-sensing autoinducer AI-2 in Escherichia coli . J Bacteriol 2005; 187:238–248 [View Article]
     [Google Scholar]
  8. Rutherford ST, Bassler BL. Bacterial quorum sensing: its role in virulence and possibilities for its control. Cold Spring Harb Perspect Med 2012; 2:a012427 [View Article]
     [Google Scholar]
  9. Calfee DP. Recent advances in the understanding and management of Klebsiella pneumoniae . F1000Res 1760; 2017:6
     [Google Scholar]
  10. Zaidi AKM, Huskins WC, Thaver D, Bhutta ZA, Abbas Z et al. Hospital-Acquired neonatal infections in developing countries. The Lancet 2005; 365:1175–1188 [View Article]
     [Google Scholar]
  11. Murphy CN, Clegg S. Klebsiella pneumoniae and type 3 fimbriae: nosocomial infection, regulation and biofilm formation. Future Microbiol 2012; 7:991–1002 [View Article]
     [Google Scholar]
  12. Paczosa MK, Mecsas J. Klebsiella pneumoniae: going on the offense with a strong defense. Microbiol Mol Biol Rev 2016; 80:629–661 [View Article]
     [Google Scholar]
  13. Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol 2004; 2:95–108 [View Article]
     [Google Scholar]
  14. Stickler DJ. Bacterial biofilms in patients with indwelling urinary catheters. Nat Clin Pract Urol 2008; 5:598–608 [View Article]
     [Google Scholar]
  15. Vuotto C, Longo F, Balice MP, Donelli G, Varaldo PE. Antibiotic resistance related to biofilm formation in Klebsiella pneumoniae . Pathogens 2014; 3:743–758 [View Article]
     [Google Scholar]
  16. Cook LC, Dunny GM. The influence of biofilms in the biology of plasmids. Microbiol Spectr 2014; 2:
     [Google Scholar]
  17. Koraimann G, Wagner MA. Social behavior and decision making in bacterial conjugation. Front Cell Infect Microbiol 2014; 4:54 [View Article]
     [Google Scholar]
  18. Barlow M. What antimicrobial resistance has taught us about horizontal gene transfer. Methods Mol Biol 2009; 532:397–411 [View Article]
     [Google Scholar]
  19. Brackman G, Coenye T. Quorum sensing inhibitors as Anti-Biofilm agents. Curr Pharm Des 2014; 21:5–11 [View Article]
     [Google Scholar]
  20. Koo H, Allan RN, Howlin RP, Stoodley P, Hall-Stoodley L. Targeting microbial biofilms: current and prospective therapeutic strategies. Nat Rev Microbiol 2017; 15:740–755 [View Article]
     [Google Scholar]
  21. Wang Y-M, Dong W-L, Odah KA, Kong L-C, Ma H-X et al. Transcriptome analysis reveals AI-2 relevant genes of multi-drug resistant Klebsiella pneumoniae in response to eugenol at Sub-MIC. Front Microbiol 2019; 10:1159 [View Article]
     [Google Scholar]
  22. Weiland-Bräuer N, Kisch MJ, Pinnow N, Liese A, Schmitz RA. Highly effective inhibition of biofilm formation by the first metagenome-derived AI-2 quenching enzyme. Front Microbiol 2016; 7:1098 [View Article]
     [Google Scholar]
  23. Zhu H, Liu H-J, Ning S-J, Gao Y-L. A luxS-dependent transcript profile of cell-to-cell communication in Klebsiella pneumoniae . Mol Biosyst 2011; 7:3164–3168 [View Article]
     [Google Scholar]
  24. De Araujo C, Balestrino D, Roth L, Charbonnel N, Forestier C. Quorum sensing affects biofilm formation through lipopolysaccharide synthesis in Klebsiella pneumoniae . Res Microbiol 2010; 161:595–603 [View Article]
     [Google Scholar]
  25. Ying J, Wu S, Zhang K, Wang Z, Zhu W et al. Comparative genomics analysis of pKF3-94 in Klebsiella pneumoniae reveals plasmid compatibility and horizontal gene transfer. Front Microbiol 2015; 6:831 [View Article]
     [Google Scholar]
  26. Taga ME, Xavier KB. Methods for analysis of bacterial Autoinducer‐2 production. Curr Protoc Microbiol 2011; 23: [View Article]
     [Google Scholar]
  27. Herring CD, Glasner JD, Blattner FR. Gene replacement without selection: regulated suppression of amber mutations in Escherichia coli . Gene 2003; 311:153–163 [View Article]
     [Google Scholar]
  28. Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 2000; 97:6640–6645 [View Article]
     [Google Scholar]
  29. Chalker AF, Minehart HW, Hughes NJ, Koretke KK, Lonetto MA et al. Systematic identification of selective essential genes in Helicobacter pylori by genome prioritization and allelic replacement mutagenesis. J Bacteriol 2001; 183:1259–1268 [View Article]
     [Google Scholar]
  30. Chang AC, Cohen SN. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 1978; 134:1141–1156 [View Article]
     [Google Scholar]
  31. Bassler BL, Greenberg EP, Stevens AM. Cross-species induction of luminescence in the quorum-sensing bacterium Vibrio harveyi . J Bacteriol 1997; 179:4043–4045 [View Article]
     [Google Scholar]
  32. Wilksch JJ, Yang J, Clements A, Gabbe JL, Short KR et al. MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression. PLoS Pathog 2011; 7:e1002204 [View Article]
     [Google Scholar]
  33. Kong EF, Tsui C, Kucharíková S, Andes D, Van Dijck P et al. Commensal protection of Staphylococcus aureus against antimicrobials by Candida albicans biofilm matrix. MBio 2016; 7: [View Article]
     [Google Scholar]
  34. Beeston AL, Surette MG. pfs-dependent regulation of autoinducer 2 production in Salmonella enterica serovar Typhimurium. J Bacteriol 2002; 184:3450–3456 [View Article]
     [Google Scholar]
  35. Zhu H, Liu H-J, Ning S-J, Gao Y-L. The response of type 2 quorum sensing in Klebsiella pneumoniae to a fluctuating culture environment. DNA Cell Biol 2012; 31:455–459 [View Article]
     [Google Scholar]
  36. Surette MG, Bassler BL. Quorum sensing in Escherichia coli and Salmonella typhimurium . Proc Natl Acad Sci U S A 1998; 95:7046–7050 [View Article]
     [Google Scholar]
  37. Kim S-P, Kim C-M, Shin S-H. Cyclic AMP and cyclic AMP-receptor protein modulate the autoinducer-2-mediated quorum sensing system in Vibrio vulnificus . Curr Microbiol 2012; 65:701–710 [View Article]
     [Google Scholar]
  38. Wang L, Hashimoto Y, Tsao C-Y, Valdes JJ, Bentley WE. Cyclic AMP (cAMP) and cAMP receptor protein influence both synthesis and uptake of extracellular autoinducer 2 in Escherichia coli . J Bacteriol 2005; 187:2066–2076 [View Article]
     [Google Scholar]
  39. Jesudhasan PR, Cepeda ML, Widmer K, Dowd SE, Soni KA et al. Transcriptome analysis of genes controlled by luxS/autoinducer-2 in Salmonella enterica serovar typhimurium. Foodborne Pathog Dis 2010; 7:399–410 [View Article]
     [Google Scholar]
  40. Yoon Y, Sofos JN. Autoinducer-2 activity of gram-negative foodborne pathogenic bacteria and its influence on biofilm formation. J Food Sci 2008; 73:M140–M147 [View Article]
     [Google Scholar]
  41. Tchieu JH, Norris V, Edwards JS, Saier MH. The complete phosphotransferase system in Escherichia coli . J Mol Microbiol Biotechnol 2001; 3:329–346
     [Google Scholar]
  42. Bandara HMHN, Lam OLT, Jin LJ, Samaranayake L. Microbial chemical signaling: a current perspective. Crit Rev Microbiol 2012; 38:217–249 [View Article]
     [Google Scholar]
  43. Dickschat JS. Quorum sensing and bacterial biofilms. Nat Prod Rep 2010; 27:343–369 [View Article]
     [Google Scholar]
  44. González Barrios AF, Zuo R, Hashimoto Y, Yang L, Bentley WE et al. Autoinducer 2 controls biofilm formation in Escherichia coli through a novel motility quorum-sensing regulator (MqsR, B3022). J Bacteriol 2006; 188:305–316 [View Article]
     [Google Scholar]
  45. Li H, Li X, Song C, Zhang Y, Wang Z et al. Autoinducer-2 facilitates Pseudomonas aeruginosa PAO1 pathogenicity in vitro and in vivo . Front Microbiol 1944; 2017:8
     [Google Scholar]
  46. Prouty AM, Schwesinger WH, Gunn JS. Biofilm formation and interaction with the surfaces of gallstones by Salmonella spp. Infect Immun 2002; 70:2640–2649 [View Article]
     [Google Scholar]
  47. Huang T-W, Lam I, Chang H-Y, Tsai S-F, Palsson BO et al. Capsule deletion via a λ-Red knockout system perturbs biofilm formation and fimbriae expression in Klebsiella pneumoniae MGH 78578. BMC Res Notes 2014; 7:13 [View Article]
     [Google Scholar]
  48. Murphy CN, Mortensen MS, Krogfelt KA, Clegg S. Role of Klebsiella pneumoniae type 1 and type 3 fimbriae in colonizing silicone tubes implanted into the bladders of mice as a model of catheter-associated urinary tract infections. Infect Immun 2013; 81:3009–3017 [View Article]
     [Google Scholar]
  49. Tan JWH, Wilksch JJ, Hocking DM, Wang N, Srikhanta YN et al. Positive autoregulation of mrkHI by the Cyclic Di-GMP-dependent MrkH protein in the biofilm regulatory circuit of Klebsiella pneumoniae . J Bacteriol 2015; 197:1659–1667
     [Google Scholar]
  50. Wang H, Wilksch JJ, Strugnell RA, Gee ML. Role of capsular polysaccharides in biofilm formation: an AFM nanomechanics study. ACS Appl Mater Interfaces 2015; 7:13007–13013 [View Article]
     [Google Scholar]
  51. Vuotto C, Longo F, Pascolini C, Donelli G, Balice MP et al. Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains. J Appl Microbiol 2017; 123:1003–1018 [View Article]
     [Google Scholar]
  52. Di Martino P, Cafferini N, Joly B, Darfeuille-Michaud A. Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. Res Microbiol 2003; 154:9–16 [View Article]
     [Google Scholar]
  53. Langstraat J, Bohse M, Clegg S. Type 3 fimbrial shaft (MrkA) of Klebsiella pneumoniae, but not the fimbrial adhesin (MrkD), facilitates biofilm formation. Infect Immun 2001; 69:5805–5812 [View Article]
     [Google Scholar]
  54. Stanley NR, Lazazzera BA. Environmental signals and regulatory pathways that influence biofilm formation. Mol Microbiol 2004; 52:917–924 [View Article]
     [Google Scholar]
  55. Lee C-H, Chen I-L, Chuah S-K, Tai W-C, Chang C-C et al. Impact of glycemic control on capsular polysaccharide biosynthesis and opsonophagocytosis of Klebsiella pneumoniae: implications for invasive syndrome in patients with diabetes mellitus. Virulence 2016; 7:770–778 [View Article]
     [Google Scholar]
  56. Lin C-T, Chen Y-C, Jinn T-R, Wu C-C, Hong Y-M et al. Role of the cAMP-dependent carbon catabolite repression in capsular polysaccharide biosynthesis in Klebsiella pneumoniae . PLoS One 2013; 8:e54430 [View Article]
     [Google Scholar]
  57. Jackson DW, Simecka JW, Romeo T. Catabolite repression of Escherichia coli biofilm formation. J Bacteriol 2002; 184:3406–3410 [View Article]
     [Google Scholar]
  58. Sutrina SL, Daniel K, Lewis M, Charles NT, Anselm CKE et al. Biofilm growth of Escherichia coli Is subject to cAMP-dependent and cAMP-independent inhibition. J Mol Microbiol Biotechnol 2015; 25:209–225 [View Article]
     [Google Scholar]
  59. Schembri MA, Dalsgaard D, Klemm P. Capsule shields the function of short bacterial adhesins. J Bacteriol 2004; 186:1249–1257 [View Article]
     [Google Scholar]
  60. Balestrino D, Ghigo J-M, Charbonnel N, Haagensen JAJ, Forestier C. The characterization of functions involved in the establishment and maturation of Klebsiella pneumoniae in vitro biofilm reveals dual roles for surface exopolysaccharides. Environ Microbiol 2008; 10:685–701 [View Article]
     [Google Scholar]
  61. Favre-Bonte S, Joly B, Forestier C. Consequences of reduction of Klebsiella pneumoniae capsule expression on interactions of this bacterium with epithelial cells. Infect Immun 1999; 67:554–561 [View Article]
     [Google Scholar]
  62. Pal S, Verma J, Mallick S, Rastogi SK, Kumar A et al. Absence of the glycosyltransferase WcaJ in Klebsiella pneumoniae ATCC13883 affects biofilm formation, increases polymyxin resistance and reduces murine macrophage activation. Microbiology 2019; 165:891–904 [View Article]
     [Google Scholar]
  63. Schembri MA, Blom J, Krogfelt KA, Klemm P. Capsule and fimbria interaction in Klebsiella pneumoniae . Infect Immun 2005; 73:4626–4633 [View Article]
     [Google Scholar]
  64. Flemming H-C, Wingender J, Szewzyk U, Steinberg P, Rice SA et al. Biofilms: an emergent form of bacterial life. Nat Rev Microbiol 2016; 14:563–575 [View Article]
     [Google Scholar]
  65. DeLisa MP, Wu CF, Wang L, Valdes JJ, Bentley WE. DNA microarray-based identification of genes controlled by autoinducer 2-stimulated quorum sensing in Escherichia coli . J Bacteriol 2001; 183:5239–5247 [View Article]
     [Google Scholar]
  66. Junges R, Salvadori G, Shekhar S, Amdal HA, Periselneris JN et al. A quorum-sensing system that regulates Streptococcus pneumoniae biofilm formation and surface polysaccharide production. mSphere ; 2017; 2
  67. Sakuragi Y, Kolter R. Quorum-sensing regulation of the biofilm matrix genes (pel) of Pseudomonas aeruginosa . J Bacteriol 2007; 189:5383–5386 [View Article]
     [Google Scholar]
  68. Choi AHK, Slamti L, Avci FY, Pier GB, Maira-Litrán T. The pgaABCD locus of Acinetobacter baumannii encodes the production of poly-beta-1-6-N-acetylglucosamine, which is critical for biofilm formation. J Bacteriol 2009; 191:5953–5963 [View Article]
     [Google Scholar]
  69. Conover MS, Sloan GP, Love CF, Sukumar N, Deora R. The Bps polysaccharide of Bordetella pertussis promotes colonization and biofilm formation in the nose by functioning as an adhesin. Mol Microbiol 2010; 77:1439–1455 [View Article]
     [Google Scholar]
  70. França A, Vilanova M, Cerca N, Pier GB. Monoclonal antibody raised against PNAG has variable effects on static S. epidermidis biofilm accumulation in vitro. Int J Biol Sci 2013; 9:518–520 [View Article]
     [Google Scholar]
  71. Little DJ, Milek S, Bamford NC, Ganguly T, DiFrancesco BR et al. The protein BpsB is a poly-β-1,6-N-acetyl-D-glucosamine deacetylase required for biofilm formation in Bordetella bronchiseptica . J Biol Chem 2015; 290:22827–22840 [View Article]
     [Google Scholar]
  72. Vuong C, Kocianova S, Voyich JM, Yao Y, Fischer ER et al. A crucial role for exopolysaccharide modification in bacterial biofilm formation, immune evasion, and virulence. J Biol Chem 2004; 279:54881–54886 [View Article]
     [Google Scholar]
  73. Chen K-M, Chiang M-K, Wang M, Ho H-C, Lu M-C et al. The role of pgaC in Klebsiella pneumoniae virulence and biofilm formation. Microb Pathog 2014; 77:89–99 [View Article]
     [Google Scholar]
  74. Skurnik D, Roux D, Pons S, Guillard T, Lu X et al. Extended-spectrum antibodies protective against carbapenemase-producing Enterobacteriaceae . J Antimicrob Chemother 2016; 71:927–935 [View Article]
     [Google Scholar]
  75. Alshalchi SA, Anderson GG. Expression of the lipopolysaccharide biosynthesis gene lpxD affects biofilm formation of Pseudomonas aeruginosa . Arch Microbiol 2015; 197:135–145 [View Article]
     [Google Scholar]
  76. Bandara HMHN, Lam OLT, Watt RM, Jin LJ, Samaranayake LP. Bacterial lipopolysaccharides variably modulate in vitro biofilm formation of Candida species. J Med Microbiol 2010; 59:1225–1234 [View Article]
     [Google Scholar]
  77. Kumar A, Mallik D, Pal S, Mallick S, Sarkar S et al. Escherichia coli O8-antigen enhances biofilm formation under agitated conditions. FEMS Microbiol Lett 2015; 362:fnv112 [View Article]
     [Google Scholar]
  78. Vogeleer P, Vincent AT, Chekabab SM, Charette SJ, Novikov A et al. Regulation of waaH by PhoB during Pi Starvation Promotes Biofilm Formation by Escherichia coli O157:H7. J Bacteriol 2019; 201: [View Article]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001148
Loading
Investigation of LuxS-mediated quorum sensing in Klebsiella pneumoniae
J. Med. Microbiol. 69, 402 (2020); https://doi.org/10.1099/jmm.0.001148
/content/journal/jmm/10.1099/jmm.0.001148
/content/journal/jmm/10.1099/jmm.0.001148
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error