1887

Abstract

the Gram-negative facultative pathogen, resides in the aquatic environment and infects humans and causes diarrhoeagenic cholera. Although the environment differs drastically, thrives in both of these conditions aptly and chitinases play a vital role in their persistence and nutrient acquisition. Chitinases also play a role in pathogenesis. Chitinases and its downstream chitin utilization genes are regulated by sensor histidine kinase ChiS, which also plays a significant role in pathogenesis. Recent exploration suggests that CytR, a transcription factor of the LacI family in also regulates chitinase secretion in environmental conditions. Since chitinases and chitinase regulator ChiS is involved in pathogenesis, CytR might also play a significant role in pathogenicity. However, the role of CytR in pathogenesis is yet to be known. This study explores the regulation of CytR on the activation of ChiS in the presence of mucin and its role in pathogenesis. Therefore, we created a CytR isogenic mutant strain of (CytR¯) and found considerably less β-hexosaminidase enzyme production, which is an indicator of ChiS activity. The CytR¯ strain greatly reduced the expression of chitinases and in mucin-supplemented media. Electron microscopy showed that the CytR¯ strain was aflagellate. The expression of flagellar-synthesis regulatory genes , and class III flagellar-synthesis genes were reduced in the CytR¯ strain. The isogenic CytR mutant showed less growth compared to the wild-type in mucin-supplemented media as well as demonstrated highly retarded motility and reduced mucin-layer penetration. The CytR mutant revealed decreased adherence to the HT-29 cell line. In animal models, reduced fluid accumulation and colonization were observed during infection with the CytR¯ strain due to reduced expression of , and . Collectively these data suggest that CytR plays an important role in pathogenesis.

Funding
This study was supported by the:
  • Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) (Award OUP 3-2)
    • Principle Award Recipient: Nabendu Sekhar Chatterjee
  • Indian Council of Medical Research (Award 3/1/3/JRF-2015/HRD-LS/106/60046/56, Dated- 10 September 2015)
    • Principle Award Recipient: Suman Das
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000949
2020-11-04
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/166/12/1136.html?itemId=/content/journal/micro/10.1099/mic.0.000949&mimeType=html&fmt=ahah

References

  1. Kaper JB, Morris JG, Levine MM. Cholera. Clin Microbiol Rev 1995; 8:48–86 [View Article]
    [Google Scholar]
  2. Morris JG. Cholera—Modern pandemic disease of ancient lineage. Emerg Infect Dis 2011; 17:2099–2104 [View Article]
    [Google Scholar]
  3. Mutreja A, Kim DW, Thomson NR, Connor TR, Lee JH et al. Evidence for several waves of global transmission in the seventh cholera pandemic. Nature 2011; 477:462–465 [View Article]
    [Google Scholar]
  4. Huq A, Small EB, West PA, Huq MI, Rahman R et al. Ecological relationships between Vibrio cholerae and planktonic crustacean copepods. Appl Environ Microbiol 1983; 45:275–283 [View Article]
    [Google Scholar]
  5. Meibom KL, Li XB, Nielsen AT, Wu CY, Roseman S et al. The Vibrio cholerae chitin utilization program. Proc Natl Acad Sci U S A 2004; 101:2524–2529 [View Article][PubMed]
    [Google Scholar]
  6. Orikoshi H, Nakayama S, Miyamoto K, Hanato C, Yasuda M et al. Roles of four chitinases (chia, ChiB, chiC, and ChiD) in the chitin degradation system of marine bacterium Alteromonas sp. strain O-7. Appl Environ Microbiol 2005; 71:1811–1815 [View Article]
    [Google Scholar]
  7. Harel J, Fairbrother J, Forget C, Desautels C, Moore J. Virulence factors associated with F165-positive Escherichia coli strains isolated from piglets and calves. Vet Microbiol 1993; 38:139–155 [View Article]
    [Google Scholar]
  8. Reddi G, Pruss K, Cottingham KL, Taylor RK, Almagro-Moreno S. Catabolism of mucus components influences motility of Vibrio cholerae in the presence of environmental reservoirs. PLoS One 2018; 13:1–16 [View Article]
    [Google Scholar]
  9. Tran HT, Barnich N, Mizoguchi E. Potential role of chitinases and chitin-binding proteins in host-microbial interactions during the development of intestinal inflammation. Histol Histopathol 2011; 26:1453–1464
    [Google Scholar]
  10. Frederiksen RF, Paspaliari DK, Larsen T, Storgaard BG, Larsen MH et al. Bacterial chitinases and chitin-binding proteins as virulence factors. Microbiology 2013; 159:833–847 [View Article]
    [Google Scholar]
  11. Mondal M, Nag D, Koley H, Saha DR, Chatterjee NS. The Vibrio cholerae extracellular chitinase ChiA2 is important for survival and pathogenesis in the host intestine. PLoS One 2014; 9:e103119 [View Article]
    [Google Scholar]
  12. Chourashi R, Mondal M, Sinha R, Debnath A, Das S et al. Role of a sensor histidine kinase ChiS of Vibrio cholerae in pathogenesis. Int J Med Microbiol 2016; 306:657–665 [View Article]
    [Google Scholar]
  13. Watve SS, Thomas J, Hammer BK. CytR is a global positive regulator of competence, type VI secretion, and chitinases in Vibrio cholerae. PLoS One 2015; 10:e0138834 [View Article]
    [Google Scholar]
  14. Price MN, Dehal PS, Arkin AP. Orthologous transcription factors in bacteria have different functions and regulate different genes. PLoS Comput Biol 2007; 3:e175–50 [View Article]
    [Google Scholar]
  15. Sernova NV, Gelfand MS. Comparative genomics of CytR, an unusual member of the lacI family of transcription factors. PLoS One 2012; 7:e44194–16 [View Article][PubMed]
    [Google Scholar]
  16. Hammer-jespersen K, Nygaard P. Multiple regulation of nucleoside catabolizing enzymes in Escherichia coli; 1976; 5549–55
  17. Antonova ES, Bernardy EE, Hammer BK. Natural competence in Vibrio cholerae is controlled by a nucleoside scavenging response that requires CytR-dependent anti-activation. Mol Microbiol 2012; 86:1215–1231 [View Article]
    [Google Scholar]
  18. Valentin‐Hansen P, Søgaard‐Andersen L, Pedersen H. A flexible partnership: The CytR anti-activator and the cAMP-CRF activator protein, comrades in transcription control. Mol Microbiol 1996; 20:461–466 [View Article]
    [Google Scholar]
  19. Silva AJ, Benitez JA. Vibrio cholerae biofilms and cholera pathogenesis. PLoS Negl Trop Dis 2016; 10:e0004330–25 [View Article]
    [Google Scholar]
  20. Conner JG, Teschler JK, Jones CJ, Yildiz FH. VMBF-0015-2015_Vibrio review.; 20161–32
  21. Nelson EJ, Harris JB, Glenn Morris J, Calderwood SB, Camilli A. Cholera transmission: the host, pathogen and bacteriophage dynamic. Nat Rev Microbiol 2009; 7:693–702 [View Article]
    [Google Scholar]
  22. Philippe N, Alcaraz JP, Coursange E, Geiselmann J, Schneider D. Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria. Plasmid 2004; 51:246–255 [View Article]
    [Google Scholar]
  23. Skorupski K, Taylor RK. Positive selection vectors for allelic exchange. Gene 1996; 169:47–52 [View Article]
    [Google Scholar]
  24. Liu Z, Wang Y, Liu S, Sheng Y, Rueggeberg KG et al. Vibrio cholerae represses polysaccharide synthesis to promote motility in mucosa. Infect Immun 2015; 83:1114–1121 [View Article]
    [Google Scholar]
  25. Herrera CM, Crofts AA, Henderson JC, Pingali SC, Davies BW et al. Virulence through endotoxin modification; 2014; 51–13
  26. Bansil R, Turner BS. Mucin structure, aggregation, physiological functions and biomedical applications. Curr Opin Colloid Interface Sci 2006; 11:164–170 [View Article]
    [Google Scholar]
  27. Vercruysse M, Köhrer C, Davies BW, Arnold MFF, Mekalanos JJ et al. The highly conserved bacterial RNase YbeY is essential in Vibrio cholerae, playing a critical role in virulence, stress regulation, and RNA processing. PLoS Pathog 2014; 10:e1004175 [View Article]
    [Google Scholar]
  28. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001; 25:402–408 [View Article]
    [Google Scholar]
  29. Li X, Roseman S. The chitinolytic cascade in vibrios is regulated by chitin oligosaccharides and a two-component chitin catabolic sensor/kinase. Proc Natl Acad Sci U S A 2004; 101:627–631 [View Article]
    [Google Scholar]
  30. Miyashiro T, Hsiao A, Liu Z, Tsou A, Zhu J et al. Mucosal penetration primes Vibrio cholerae for host colonization by repressing quorum sensing. Proc Natl Acad Sci 2008; 105:9769–9774
    [Google Scholar]
  31. Yeung ATY, Parayno A, Hancock REW. Mucin promotes rapid surface motility in; 2012; 31–12
  32. Zheng J, Shin OS, Cameron DE, Mekalanos JJ. Quorum sensing and a global regulator TsrA control expression of type VI secretion and virulence in Vibrio cholerae . Proc Natl Acad Sci U S A 2010; 107:21128–21133 [View Article]
    [Google Scholar]
  33. Koley H, Mitra R, Basu A, Mukhopadhyay AK, Saha PK et al. Response of wild-type mutants of Vibrio cholerae O1 possessing different combinations of virulence genes in the ligated rabbit ileal loop and in Ussing chambers: evidence for the presence of additional secretogen. J Med Microbiol 1999; 48:51–57 [View Article]
    [Google Scholar]
  34. Holmgren J. Comparison of the tissue receptors for Vibrio cholerae and Escherichia coli enterotoxins by means of gangliosides and natural cholera toxoid. Infect Immun 1973; 8:851–859 [View Article]
    [Google Scholar]
  35. De K, Ramamurthy T, Faruque SM, Yamasaki S, Takeda Y et al. Molecular characterisation of rough strains of Vibrio cholerae isolated from diarrhoeal cases in India and their comparison to smooth strains. FEMS Microbiol Lett 2004; 232:23–30 [View Article]
    [Google Scholar]
  36. Patra T, Koley H, Ramamurthy T, Ghose AC, Nandy RK. The Entner-Doudoroff pathway is obligatory for gluconate utilization and contributes to the pathogenicity of Vibrio cholerae . J Bacteriol 2012; 194:3377–3385 [View Article]
    [Google Scholar]
  37. Bjork S, Breimer ME, Hansson GC, Karlsson K, Leffler H. Structures of blood group glycosphingolipids of human small intestine. J Biol Chem 1987; 262:6758–6765
    [Google Scholar]
  38. Chen H, Chang CC, Mau WJ, Yen LS. Evaluation of N-acetylchitooligosaccharides as the main carbon sources for the growth of intestinal bacteria. FEMS Microbiol Lett 2002; 209:51–54 [View Article]
    [Google Scholar]
  39. Alemka A, Corcionivoschi N, Bourke B. Defense and adaptation: the complex inter-relationship between Campylobacter jejuni and mucus. Front Cell Infect Microbiol 2012; 2:15 [View Article]
    [Google Scholar]
  40. Fabich AJ, Jones SA, Chowdhury FZ, Cernosek A, Anderson A et al. Comparison of carbon nutrition for pathogenic and commensal Escherichia coli strains in the mouse intestine. Infect Immun 2008; 76:1143–1152 [View Article]
    [Google Scholar]
  41. KM N, Ferreyra JA, Higginbottom SK, Lynch JB, Kashyap PC et al. Microbiota-liberated host sugars facilitate post-antibiotic expansion of enteric pathogens. Nature 2013; 502:96–99
    [Google Scholar]
  42. Ghosh S, Rao KH, Sengupta M, Bhattacharya SK, Datta A. Two gene clusters co-ordinate for a functional N-acetylglucosamine catabolic pathway in Vibrio cholerae . Mol Microbiol 2011; 80:1549–1560 [View Article]
    [Google Scholar]
  43. Naseem S, Konopka JB. N-acetylglucosamine regulates virulence properties in microbial pathogens. PLoS Pathog 2015; 11:e1004947–11 [View Article]
    [Google Scholar]
  44. Echazarreta MA, Klose KE. Vibrio flagellar synthesis. Front Cell Infect Microbiol 2019; 9:1–11 [View Article]
    [Google Scholar]
  45. Klose KE, Mekalanos JJ. Distinct roles of an alternative sigma factor during both free‐swimming and colonizing phases of the Vibrio cholerae pathogenic cycle. Mol Microbiol 1998; 28:501–520 [View Article]
    [Google Scholar]
  46. Prouty MG, Correa NE, Klose KE. The novel sigma54- and sigma28-dependent flagellar gene transcription hierarchy of Vibrio cholerae. Mol Microbiol 2001; 39:1595–1609 [View Article]
    [Google Scholar]
  47. Syed KA, Beyhan S, Correa N, Queen J, Liu J et al. The Vibrio cholerae flagellar regulatory hierarchy controls expression of virulence factors. J Bacteriol 2009; 191:6555–6570 [View Article]
    [Google Scholar]
  48. Correa NE, Lauriano CM, McGee R, Klose KE. Phosphorylation of the flagellar regulatory protein FlrC is necessary for Vibrio cholerae motility and enhanced colonization. Mol Microbiol 2000; 35:743–755 [View Article]
    [Google Scholar]
  49. Krebs SJ, Taylor RK. Protection and attachment of Vibrio cholerae mediated by the toxin-coregulated pilus in the infant mouse model. J Bacteriol 2011; 193:5260–5270 [View Article]
    [Google Scholar]
  50. Bhowmick R, Ghosal A, Das B, Koley H, Saha DR et al. Intestinal adherence of Vibrio cholerae involves a coordinated interaction between colonization factor GbpA and mucin. Infect Immun 2008; 76:4968–4977 [View Article]
    [Google Scholar]
  51. Matson JS, Withey JH, DiRita VJ. Regulatory networks controlling Vibrio cholerae virulence gene expression. Infect Immun 2007; 75:5542–5549 [View Article]
    [Google Scholar]
  52. Childers BM, Klose KE. Regulation of virulence in Vibrio cholerae : the ToxR regulon. Future Microbiol 2007; 2:335–344 [View Article]
    [Google Scholar]
  53. Kovacikova G, Lin W, Skorupski K. Vibrio cholerae AphA uses a novel mechanism for virulence gene activation that involves interaction with the LysR-type regulator AphB at the tcpPH promoter. Mol Microbiol 2004; 53:129–142 [View Article]
    [Google Scholar]
  54. Skorupski K, Taylor RK. Cyclic AMP and its receptor protein negatively regulate the coordinate expression of cholera toxin and toxin-coregulated pilus in Vibrio cholerae. Proc Natl Acad Sci U S A 1997; 94:265–270 [View Article]
    [Google Scholar]
  55. Taylor RK, Miller VL, Furlong DB, Mekalanos JJ. Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin. Proc Natl Acad Sci U S A 1987; 84:2833–2837 [View Article]
    [Google Scholar]
  56. Ritchie JM, Rui H, Bronson RT, Waldor MK. Back to the future: studying cholera pathogenesis using infant rabbits. MBio 2010; 1:1–13 [View Article]
    [Google Scholar]
  57. Kim SY, Thanh XTT, Jeong K, Kim SB, Pan SO et al. Contribution of six flagellin genes to the flagellum biogenesis of Vibrio vulnificus and in vivo invasion. Infect Immun 2014; 82:29–42 [View Article]
    [Google Scholar]
  58. Meron D, Efrony R, Johnson WR, Schaefer AL, Morris PJ et al. Role of flagella in virulence of the coral pathogen Vibrio coralliilyticus. Appl Environ Microbiol 2009; 75:5704–5707 [View Article]
    [Google Scholar]
  59. McGee K, Hörstedt P, Milton DL. Identification and characterization of additional flagellin genes from Vibrio anguillarum. J Bacteriol 1996; 178:5188–5198 [View Article]
    [Google Scholar]
  60. Ormonde P, Hörstedt P, O'Toole R, Milton DL. Role of motility in adherence to and invasion of a fish cell line by Vibrio anguillarum. J Bacteriol 2000; 182:2326–2328 [View Article]
    [Google Scholar]
  61. Lee SH, Butler SM, Camilli A. Selection for in vivo regulators of bacterial virulence. Proc Natl Acad Sci USA 2001; 98:6889–6894 [View Article]
    [Google Scholar]
  62. Matsumoto H, Muroi H, Umehara M, Yoshitake Y, Tsuyumu S. S. Peh production flagellum synthesis, and virulence reduced in Erwinia carotovora subsp. carotovora by mutation in a homologue of cytR. Mol Plant-Microbe Interact 2003; 16:389–397 [View Article]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000949
Loading
/content/journal/micro/10.1099/mic.0.000949
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
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