1887

Abstract

Enterohaemorrhagic (EHEC) produces Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2). Although and were found within the late operons of the Stx-encoding phages (Stx-phages), could mainly be transcribed from the promoter ( ), which represents the functional operator-binding site (Fur box) for the transcriptional regulator Fur (ferric uptake regulator), upstream of . In this study, we found that the production of Stx1 by EHEC was affected by oxygen concentration. Increased Stx1 production in the presence of oxygen is dependent on Fur, which is an Fe-responsive transcription factor. The intracellular Fe pool was lower under microaerobic conditions than under anaerobic conditions, suggesting that lower Fe availability drove the formation of less Fe-Fur, less DNA binding to the region, and an increase in Stx1 production.

Funding
This study was supported by the:
  • Japan Agency for Medical Research and Development (Award JP18jk021007)
    • Principle Award Recipient: TakeshiShimizu
  • Japan Society for the Promotion of Science London (Award 20K07474)
    • Principle Award Recipient: TakeshiShimizu
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.001122
2021-12-24
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/167/12/mic001122.html?itemId=/content/journal/micro/10.1099/mic.0.001122&mimeType=html&fmt=ahah

References

  1. Jackson MP, Neill RJ, O’Brien AD, Holmes RK, Newland JW. Nucleotide sequence analysis and comparison of the structural genes for Shiga-like toxin I and Shiga-like toxin II encoded by bacteriophages from Escherichia coli 933. FEMS Microbiol Lett 1987; 44:109–114 [View Article]
    [Google Scholar]
  2. Tesh VL, O’Brien AD. The pathogenic mechanisms of Shiga toxin and the Shiga-like toxins. Mol Microbiol 1991; 5:1817–1822 [View Article] [PubMed]
    [Google Scholar]
  3. Neely MN, Friedman DI. Functional and genetic analysis of regulatory regions of coliphage H-19B: location of shiga-like toxin and lysis genes suggest a role for phage functions in toxin release. Mol Microbiol 1998; 28:1255–1267 [View Article] [PubMed]
    [Google Scholar]
  4. Plunkett G, Durfee TJ, Blattner FR. Sequence of Shiga toxin 2 phage 933W from Escherichia coli O157:H7: Shiga toxin as a phage late-gene product. J Bacteriol 1999; 181:1767–1778 [View Article] [PubMed]
    [Google Scholar]
  5. Calderwood SB, Auclair F, Donohue-Rolfe A, Keusch GT, Mekalanos JJ. Nucleotide sequence of the Shiga-like toxin genes of Escherichia coli. Proc Natl Acad Sci U S A 1987; 84:4364–4368 [View Article] [PubMed]
    [Google Scholar]
  6. Calderwood SB, Mekalanos JJ. Iron regulation of Shiga-like toxin expression in Escherichia coli is mediated by the fur locus. J Bacteriol 1987; 169:4759–4764 [View Article] [PubMed]
    [Google Scholar]
  7. Head SC, Petric M, Richardson S, Roscoe M, Karmali MA. Purification and characterization of verocytotoxin 2. FEMS Microbiol Lett 1988; 51:211–215 [View Article]
    [Google Scholar]
  8. De Grandis S, Ginsberg J, Toone M, Climie S, Friesen J et al. Nucleotide sequence and promoter mapping of the Escherichia coli Shiga-like toxin operon of bacteriophage H-19B. J Bacteriol 1987; 169:4313–4319 [View Article] [PubMed]
    [Google Scholar]
  9. Hull AE, Acheson DW, Echeverria P, Donohue-Rolfe A, Keusch GT. Mitomycin immunoblot colony assay for detection of Shiga-like toxin-producing Escherichia coli in fecal samples: comparison with DNA probes. J Clin Microbiol 1993; 31:1167–1172 [View Article] [PubMed]
    [Google Scholar]
  10. Shimizu T, Ohta Y, Noda M. Shiga toxin 2 is specifically released from bacterial cells by two different mechanisms. Infect Immun 2009; 77:2813–2823 [View Article] [PubMed]
    [Google Scholar]
  11. Ichimura K, Shimizu T, Matsumoto A, Hirai S, Yokoyama E et al. Nitric oxide-enhanced Shiga toxin production was regulated by Fur and RecA in enterohemorrhagic Escherichia coli O157. Microbiologyopen 2017; 6: [View Article] [PubMed]
    [Google Scholar]
  12. Porcheron G, Dozois CM. Interplay between iron homeostasis and virulence: Fur and RyhB as major regulators of bacterial pathogenicity. Vet Microbiol 2015; 179:2–14 [View Article] [PubMed]
    [Google Scholar]
  13. Hantke K. Iron and metal regulation in bacteria. Curr Opin Microbiol 2001; 4:172–177 [View Article] [PubMed]
    [Google Scholar]
  14. Beauchene NA, Myers KS, Chung D, Park DM, Weisnicht AM et al. Impact of anaerobiosis on expression of the iron-responsive Fur and RyhB regulons. mBio 2015; 6:e01947–15 [View Article] [PubMed]
    [Google Scholar]
  15. Wagner PL, Waldor MK. Bacteriophage control of bacterial virulence. Infect Immun 2002; 70:3985–3993 [View Article] [PubMed]
    [Google Scholar]
  16. Vareille M, de Sablet T, Hindré T, Martin C, Gobert AP. Nitric oxide inhibits Shiga-toxin synthesis by enterohemorrhagic Escherichia coli. Proc Natl Acad Sci U S A 2007; 104:10199–10204 [View Article] [PubMed]
    [Google Scholar]
  17. Albenberg L, Esipova TV, Judge CP, Bittinger K, Chen J et al. Correlation between intraluminal oxygen gradient and radial partitioning of intestinal microbiota. Gastroenterology 2014; 147:1055–1063 [View Article]
    [Google Scholar]
  18. Marteyn B, West NP, Browning DF, Cole JA, Shaw JG et al. Modulation of Shigella virulence in response to available oxygen in vivo. Nature 2010; 465:355–358 [View Article] [PubMed]
    [Google Scholar]
  19. Marteyn B, Scorza FB, Sansonetti PJ, Tang C. Breathing life into pathogens: the influence of oxygen on bacterial virulence and host responses in the gastrointestinal tract. Cell Microbiol 2011; 13:171–176 [View Article] [PubMed]
    [Google Scholar]
  20. Gunsalus RP, Park SJ. Aerobic-anaerobic gene regulation in Escherichia coli: control by the ArcAB and Fnr regulons. Res Microbiol 1994; 145:437–450 [View Article] [PubMed]
    [Google Scholar]
  21. Sawers G. The aerobic/anaerobic interface. Curr Opin Microbiol 1999; 2:181–187 [View Article] [PubMed]
    [Google Scholar]
  22. Beauchene NA, Mettert EL, Moore LJ, Keleş S, Willey ER et al. O2 availability impacts iron homeostasis in Escherichia coli. Proc Natl Acad Sci U S A 2017; 114:12261–12266 [View Article] [PubMed]
    [Google Scholar]
  23. Kaplan J, Ward DM. The essential nature of iron usage and regulation. Curr Biol 2013; 23:R642–6 [View Article] [PubMed]
    [Google Scholar]
  24. Shimizu T, Kawakami S, Sato T, Sasaki T, Higashide M et al. The serine 31 residue of the B subunit of Shiga toxin 2 is essential for secretion in enterohemorrhagic Escherichia coli. Infect Immun 2007; 75:2189–2200 [View Article] [PubMed]
    [Google Scholar]
  25. Noda M, Yutsudo T, Nakabayashi N, Hirayama T, Takeda Y. Purification and some properties of Shiga-like toxin from Escherichia coli O157:H7 that is immunologically identical to Shiga toxin. Microb Pathog 1987; 2:339–349 [View Article] [PubMed]
    [Google Scholar]
  26. Imlay JA, Linn S. Bimodal pattern of killing of DNA-repair-defective or anoxically grown Escherichia coli by hydrogen peroxide. J Bacteriol 1986; 166:519–527 [View Article] [PubMed]
    [Google Scholar]
  27. Shimizu T, Matsumoto A, Noda M. Cooperative roles of nitric oxide-metabolizing enzymes to counteract nitrosative stress in enterohemorrhagic Escherichia coli. Infect Immun 2019; 87:e00334–00319 [View Article] [PubMed]
    [Google Scholar]
  28. Hayashi T, Makino K, Ohnishi M, Kurokawa K, Ishii K et al. Complete genome sequence of enterohemorrhagic Escherichia coli O157:H7 and genomic comparison with a laboratory strain K-12. DNA Res 2001; 8:11–22 [View Article] [PubMed]
    [Google Scholar]
  29. Shimizu T, Tsutsuki H, Matsumoto A, Nakaya H, Noda M. The nitric oxide reductase of enterohaemorrhagic Escherichia coli plays an important role for the survival within macrophages. Mol Microbiol 2012; 85:492–512 [View Article] [PubMed]
    [Google Scholar]
  30. Spiro S, Guest JR. FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev 1990; 6:399–428 [View Article] [PubMed]
    [Google Scholar]
  31. Calderwood SB, Mekalanos JJ. Confirmation of the Fur operator site by insertion of a synthetic oligonucleotide into an operon fusion plasmid. J Bacteriol 1988; 170:1015–1017 [View Article] [PubMed]
    [Google Scholar]
  32. Lau CKY, Krewulak KD, Vogel HJ. Bacterial ferrous iron transport: the Feo system. FEMS Microbiol Rev 2016; 40:273–298 [View Article] [PubMed]
    [Google Scholar]
  33. He G, Shankar RA, Chzhan M, Samouilov A, Kuppusamy P et al. Noninvasive measurement of anatomic structure and intraluminal oxygenation in the gastrointestinal tract of living mice with spatial and spectral EPR imaging. Proc Natl Acad Sci U S A 1999; 96:4586–4591 [View Article] [PubMed]
    [Google Scholar]
  34. Robinson CM, Sinclair JF, Smith MJ, O’Brien AD. Shiga toxin of enterohemorrhagic Escherichia coli type O157:H7 promotes intestinal colonization. Proc Natl Acad Sci U S A 2006; 103:9667–9672 [View Article] [PubMed]
    [Google Scholar]
  35. Schüller S, Phillips AD. Microaerobic conditions enhance type III secretion and adherence of enterohaemorrhagic Escherichia coli to polarized human intestinal epithelial cells. Environ Microbiol 2010; 12:2426–2435 [View Article] [PubMed]
    [Google Scholar]
  36. Carlson-Banning Km, Sperandio V. Catabolite and oxygen regulation of enterohemorrhagic Escherichia coli virulence. mbio 2016; 7:e01852-16 [View Article] [PubMed]
    [Google Scholar]
  37. Shimizu T, Ichimura K, Noda M. The surface sensor NlpE of enterohemorrhagic Escherichia coli contributes to regulation of the type III secretion system and flagella by the Cpx response to adhesion. Infect Immun 2016; 84:537–549 [View Article] [PubMed]
    [Google Scholar]
  38. Mahajan A, Currie CG, Mackie S, Tree J, McAteer S et al. An investigation of the expression and adhesin function of H7 flagella in the interaction of Escherichia coli O157 : H7 with bovine intestinal epithelium. Cell Microbiol 2009; 11:121–137 [View Article] [PubMed]
    [Google Scholar]
  39. Fink RC, Evans MR, Porwollik S, Vazquez-Torres A, Jones-Carson J et al. FNR is a global regulator of virulence and anaerobic metabolism in Salmonella enterica serovar Typhimurium (ATCC 14028s). J Bacteriol 2007; 189:2262–2273 [View Article] [PubMed]
    [Google Scholar]
  40. Delany I, Spohn G, Pacheco A-BF, Ieva R, Alaimo C et al. Autoregulation of Helicobacter pylori Fur revealed by functional analysis of the iron-binding site. Mol Microbiol 2002; 46:1107–1122 [View Article] [PubMed]
    [Google Scholar]
  41. De Lorenzo V, Herrero M, Giovannini F, Neilands JB. Fur (ferric uptake regulation) protein and CAP (catabolite-activator protein) modulate transcription of fur gene in Escherichia coli. Eur J Biochem 1988; 173:537–546 [View Article] [PubMed]
    [Google Scholar]
  42. Hernández JA, Muro-Pastor AM, Flores E, Bes MT, Peleato ML et al. Identification of a furA cis antisense RNA in the cyanobacterium Anabaena sp. PCC 7120. J Mol Biol 2006; 355:325–334 [View Article] [PubMed]
    [Google Scholar]
  43. Sala C, Forti F, Di Florio E, Canneva F, Milano A et al. Mycobacterium tuberculosis FurA autoregulates its own expression. J Bacteriol 2003; 185:5357–5362 [View Article] [PubMed]
    [Google Scholar]
  44. Boulette ML, Payne SM. Anaerobic regulation of Shigella flexneri virulence: ArcA regulates Fur and iron acquisition genes. J Bacteriol 2007; 189:6957–6967 [View Article] [PubMed]
    [Google Scholar]
  45. Runyen-Janecky LJ, Reeves SA, Gonzales EG, Payne SM. Contribution of the Shigella flexneri Sit, Iuc, and Feo iron acquisition systems to iron acquisition in vitro and in cultured cells. Infect Immun 2003; 71:1919–1928 [View Article] [PubMed]
    [Google Scholar]
  46. Sabri M, Léveillé S, Dozois CM. A SitABCD homologue from an avian pathogenic Escherichia coli strain mediates transport of iron and manganese and resistance to hydrogen peroxide. Microbiology (Reading) 2006; 152:745–758 [View Article] [PubMed]
    [Google Scholar]
  47. Becker S, Holighaus G, Gabrielczyk T, Unden G. O2 as the regulatory signal for FNR-dependent gene regulation in Escherichia coli. J Bacteriol 1996; 178:4515–4521 [View Article] [PubMed]
    [Google Scholar]
  48. Fu HA, Iuchi S, Lin EC. The requirement of ArcA and Fnr for peak expression of the cyd operon in Escherichia coli under microaerobic conditions. Mol Gen Genet 1991; 226:209–213 [View Article] [PubMed]
    [Google Scholar]
  49. Perna NT, Plunkett G 3rd, Burland V, Mau B, Glasner JD et al. Genome sequence of enterohaemorrhagic Escherichia coli O157:H7. Nature 2001; 409:529–533 [View Article] [PubMed]
    [Google Scholar]
  50. Elliott SJ, Kaper JB. Role of type 1 fimbriae in EPEC infections. Microb Pathog 1997; 23:113–118 [View Article] [PubMed]
    [Google Scholar]
  51. Shimizu T, Ohta Y, Tsutsuki H, Noda M. Construction of a novel bioluminescent reporter system for investigating Shiga toxin expression of enterohemorrhagic Escherichia coli. Gene 2011; 478:1–10 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.001122
Loading
/content/journal/micro/10.1099/mic.0.001122
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