Site-directed mutagenesis of Campylobacter concisus respiratory genes provides insight into the pathogen's growth requirements.

Stéphane L Benoit, Robert J Maier
Author Information
  1. Stéphane L Benoit: Department of Microbiology, University of Georgia, Athens, 30602, Georgia. stefbens@uga.edu.
  2. Robert J Maier: Department of Microbiology, University of Georgia, Athens, 30602, Georgia.

Abstract

Campylobacter concisus is an emerging human pathogen found throughout the entire human oral-gastrointestinal tract. The ability of C. concisus to colonize diverse niches of the human body indicates the pathogen is metabolically versatile. C. concisus is able to grow under both anaerobic conditions and microaerophilic conditions. Hydrogen (H) has been shown to enhance growth and may even be required. Analysis of several C. concisus genome sequences reveals the presence of two sets of genes encoding for distinct hydrogenases: a H-uptake-type ("Hyd") complex and a H-evolving hydrogenase ("Hyf"). Whole cells hydrogenase assays indicate that the former (H-uptake) activity is predominant in C. concisus, with activity among the highest we have found for pathogenic bacteria. Attempts to generate site-directed chromosomal mutants were partially successful, as we could disrupt hyfB, but not hydB, suggesting that H-uptake, but not H-evolving activity, is an essential respiratory pathway in C. concisus. Furthermore, the tetrathionate reductase ttrA gene was inactivated in various C. concisus genomospecies. Addition of tetrathionate to the medium resulted in a ten-fold increase in cell yield for the WT, while it had no effect on the ttrA mutant growth. To our knowledge, this is the first report of mutants in C. concisus.

References

  1. BMC Microbiol. 2011 Mar 15;11:53 [PMID: 21406111]
  2. Diagn Microbiol Infect Dis. 1986 Sep;5(3):269-72 [PMID: 3757477]
  3. PLoS One. 2014 Oct 10;9(10):e110187 [PMID: 25303479]
  4. J Biol Inorg Chem. 2004 Oct;9(7):791-9 [PMID: 15311335]
  5. FEMS Immunol Med Microbiol. 2011 Dec;63(3):387-96 [PMID: 22092566]
  6. N Engl J Med. 1969 Jul 17;281(3):122-7 [PMID: 5790483]
  7. Microbiology (Reading). 2015 Aug;161(8):1600-1612 [PMID: 26002953]
  8. Front Cell Infect Microbiol. 2012 Apr 02;2:45 [PMID: 22919636]
  9. J Clin Microbiol. 2010 Aug;48(8):2965-7 [PMID: 20519479]
  10. Eur J Gastroenterol Hepatol. 2005 Oct;17(10):1019-24 [PMID: 16148545]
  11. Mol Microbiol. 2000 Sep;37(5):1066-74 [PMID: 10972825]
  12. Nature. 2000 Feb 10;403(6770):665-8 [PMID: 10688204]
  13. Clin Infect Dis. 2007 Jul 1;45(1):29-38 [PMID: 17554697]
  14. mBio. 2014 Jan 21;5(1):e01016-13 [PMID: 24449753]
  15. Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):E3948-56 [PMID: 25157147]
  16. J Microbiol. 2010 Dec;48(6):778-83 [PMID: 21221934]
  17. J Bacteriol. 2003 Apr;185(8):2680-2 [PMID: 12670994]
  18. BMC Genomics. 2013 Aug 28;14:585 [PMID: 23984967]
  19. Int J Microbiol. 2014;2014:476047 [PMID: 25214843]
  20. Science. 2002 Nov 29;298(5599):1788-90 [PMID: 12459589]
  21. Nature. 2010 Sep 23;467(7314):426-9 [PMID: 20864996]
  22. Microbiology (Reading). 1997 Nov;143 ( Pt 11):3633-3647 [PMID: 9387241]
  23. Gut Pathog. 2016 Sep 22;8:44 [PMID: 27688814]
  24. Infect Immun. 2005 Sep;73(9):5311-8 [PMID: 16113246]
  25. Scand J Infect Dis. 2002;34(4):248-52 [PMID: 12064685]
  26. Front Microbiol. 2017 Dec 01;8:2391 [PMID: 29250055]
  27. Infect Immun. 2015 Jan;83(1):311-6 [PMID: 25368112]
  28. Biochemistry. 2000 Dec 26;39(51):16213-9 [PMID: 11123951]
  29. Microb Pathog. 2009 Jul;47(1):8-15 [PMID: 19397993]
  30. J Bacteriol. 2004 Jan;186(2):580-7 [PMID: 14702328]
  31. J Med Microbiol. 2016 Mar;65(3):219-226 [PMID: 26698172]
  32. FEMS Microbiol Lett. 1996 Jul 15;141(1):71-6 [PMID: 8764511]
  33. Mol Microbiol. 2018 May;108(4):379-396 [PMID: 29498770]
  34. J Clin Microbiol. 1989 Aug;27(8):1775-81 [PMID: 2768465]
  35. Biochem J. 2018 Apr 16;475(7):1353-1370 [PMID: 29555844]
  36. Microb Pathog. 2004 Mar;36(3):153-7 [PMID: 14726233]
  37. Trends Microbiol. 2008 Mar;16(3):107-14 [PMID: 18280160]
  38. Adv Microb Physiol. 2016;68:433-507 [PMID: 27134027]
  39. PLoS One. 2012;7(5):e38217 [PMID: 22666490]
  40. Front Cell Infect Microbiol. 2012 Feb 03;2:4 [PMID: 22919596]
  41. J Lab Clin Med. 1975 Apr;85(4):546-55 [PMID: 1120927]
  42. ISME J. 2013 Jul;7(7):1354-66 [PMID: 23466701]
  43. Gene. 1990 Sep 28;94(1):23-8 [PMID: 2227449]
  44. PLoS One. 2011;6(9):e25417 [PMID: 21966525]
  45. Infect Immun. 2004 Nov;72(11):6294-9 [PMID: 15501756]
  46. Sci Rep. 2016 Dec 02;6:38442 [PMID: 27910936]
  47. Appl Environ Microbiol. 2008 Apr;74(7):2095-102 [PMID: 18245237]
  48. Gut. 2017 Feb;66(2):235-249 [PMID: 26508508]
  49. J Bacteriol. 2009 Aug;191(16):5293-300 [PMID: 19525346]
  50. Appl Environ Microbiol. 2001 Oct;67(10):4657-61 [PMID: 11571169]
  51. Biochim Biophys Acta. 2009 Jun;1787(6):646-56 [PMID: 19171117]
  52. Gut Pathog. 2011 Oct 13;3:15 [PMID: 21992484]
  53. Mol Microbiol. 1990 Feb;4(2):231-43 [PMID: 2187144]
  54. Biochem Soc Trans. 2005 Feb;33(Pt 1):83-5 [PMID: 15667272]
  55. Microbiology (Reading). 2012 Aug;158(Pt 8):2204-2212 [PMID: 22628482]
  56. Eur J Biochem. 1992 May 15;206(1):93-102 [PMID: 1587288]
  57. EcoSal Plus. 2016 Oct;7(1): [PMID: 27735784]

MeSH Term

Campylobacter
Campylobacter Infections
Diarrhea
Gastroenteritis
Gastrointestinal Tract
Genome, Bacterial
Humans
Mutagenesis, Site-Directed
Mutation

Word Cloud

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