1887

Abstract

Purpose. The gastrointestinal tract is home to thousands of commensal bacterial species. Therefore, competition for nutrients is paramount for successful bacterial pathogen invasion of intestinal ecosystems. The human pathogen Vibrio cholerae, the causative agent of the severe diarrhoeal disease, cholera, is able to colonize the small intestine, which is protected by mucus. However, it is unclear which metabolic pathways or nutrients V. cholerae utilizes during intestinal colonization and growth.

Methodology. In this study, we investigated the effect of various metabolic key genes, including those involved in the gluconeogenesis pathway, on V. cholerae physiology and in vivo colonization.

Results. We found that gluconeogenesis is important for infant mouse colonization. Growth assays showed that mutations in the key components of gluconeogenesis pathway, PpsA and PckA, lead to a growth defect in a minimal medium supplemented with mucin as a carbon source. Furthermore, the ppsA/pckA mutants colonized poorly in the adult mouse intestine, particularly when more gut commensal flora are present.

Conclusion. Gluconeogenesis biosynthesis is important for the successful colonization of V. cholerae in a niche that is full of competing microbiota.

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2018-09-24
2024-12-05
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References

  1. Sack DA, Sack RB, Nair GB, Siddique AK. Cholera. The Lancet 2004; 363:223–233 [View Article]
    [Google Scholar]
  2. Wang H, Naseer N, Chen Y, Zhu AY, Kuai X et al. OxyR2 modulates OxyR1 activity and Vibrio cholerae oxidative stress response. Infect Immun 2017; 85:e00929-16 [View Article][PubMed]
    [Google Scholar]
  3. Krukonis ES, Dirita VJ. From motility to virulence: sensing and responding to environmental signals in Vibrio cholerae. Curr Opin Microbiol 2003; 6:186–190 [View Article][PubMed]
    [Google Scholar]
  4. Reidl J, Klose KE. Vibrio cholerae and cholera: out of the water and into the host. FEMS Microbiol Rev 2002; 26:125–139 [View Article][PubMed]
    [Google Scholar]
  5. Freter R. Probiotics the Scientific Basis 1992 pp. 111–144
    [Google Scholar]
  6. Conway T, Cohen PS. Commensal and pathogenic Escherichia coli metabolism in the gut. Microbiol Spectr 2015; 3: [View Article][PubMed]
    [Google Scholar]
  7. 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][PubMed]
    [Google Scholar]
  8. Freter R, Brickner H, Botney M, Cleven D, Aranki A. Mechanisms that control bacterial populations in continuous-flow culture models of mouse large intestinal flora. Infect Immun 1983; 39:676–685[PubMed]
    [Google Scholar]
  9. Zhao W, Caro F, Robins W, Mekalanos JJ. Antagonism toward the intestinal microbiota and its effect on Vibrio cholerae virulence. Science 2018; 359:210–213 [View Article][PubMed]
    [Google Scholar]
  10. Hsiao A, Ahmed AM, Subramanian S, Griffin NW, Drewry LL et al. Members of the human gut microbiota involved in recovery from Vibrio cholerae infection. Nature 2014; 515:423–426 [View Article][PubMed]
    [Google Scholar]
  11. Sauer U, Eikmanns BJ. The PEP-pyruvate-oxaloacetate node as the switch point for carbon flux distribution in bacteria. FEMS Microbiol Rev 2005; 29:765–794 [View Article][PubMed]
    [Google Scholar]
  12. Niersbach M, Kreuzaler F, Geerse RH, Postma PW, Hirsch HJ. Cloning and nucleotide sequence of the Escherichia coli K-12 ppsA gene, encoding PEP synthase. Mol Gen Genet 1992; 231:332–336[PubMed]
    [Google Scholar]
  13. Medina V, Pontarollo R, Glaeske D, Tabel H, Goldie H. Sequence of the pckA gene of Escherichia coli K-12: relevance to genetic and allosteric regulation and homology of E. coli phosphoenolpyruvate carboxykinase with the enzymes from Trypanosoma brucei and Saccharomyces cerevisiae. J Bacteriol 1990; 172:7151–7156 [View Article][PubMed]
    [Google Scholar]
  14. Goldie AH, Sanwal BD. Genetic and physiological characterization of Escherichia coli mutants deficient in phosphoenolpyruvate carboxykinase activity. J Bacteriol 1980; 141:1115–1121[PubMed]
    [Google Scholar]
  15. Joelsson A, Liu Z, Zhu J. Genetic and phenotypic diversity of quorum-sensing systems in clinical and environmental isolates of Vibrio cholerae. Infect Immun 2006; 74:1141–1147 [View Article][PubMed]
    [Google Scholar]
  16. Cameron DE, Urbach JM, Mekalanos JJ. A defined transposon mutant library and its use in identifying motility genes in Vibrio cholerae. Proc Natl Acad Sci USA 2008; 105:8736–8741 [View Article][PubMed]
    [Google Scholar]
  17. Metcalf WW, Jiang W, Daniels LL, Kim SK, Haldimann A et al. Conditionally replicative and conjugative plasmids carrying lacZ alpha for cloning, mutagenesis, and allele replacement in bacteria. Plasmid 1996; 35:1–13 [View Article][PubMed]
    [Google Scholar]
  18. Liu Z, Yang M, Peterfreund GL, Tsou AM, Selamoglu N et al. Vibrio cholerae anaerobic induction of virulence gene expression is controlled by thiol-based switches of virulence regulator AphB. Proc Natl Acad Sci USA 2011; 108:810–815 [View Article][PubMed]
    [Google Scholar]
  19. Hammer BK, Bassler BL. Regulatory small RNAs circumvent the conventional quorum sensing pathway in pandemic Vibrio cholerae. Proc Natl Acad Sci USA 2007; 104:11145–11149 [View Article][PubMed]
    [Google Scholar]
  20. Gardel CL, Mekalanos JJ. Alterations in Vibrio cholerae motility phenotypes correlate with changes in virulence factor expression. Infect Immun 1996; 64:2246–2255[PubMed]
    [Google Scholar]
  21. Liu Z, Wang H, Zhou Z, Sheng Y, Naseer N et al. Thiol-based switch mechanism of virulence regulator AphB modulates oxidative stress response in Vibrio cholerae. Mol Microbiol 2016; 102:939–949 [View Article][PubMed]
    [Google Scholar]
  22. Ni J, Shen TD, Chen EZ, Bittinger K, Bailey A et al. A role for bacterial urease in gut dysbiosis and Crohn's disease. Sci Transl Med 2017; 9:eaah6888 [View Article][PubMed]
    [Google Scholar]
  23. Miller JH. Experiments in Molecular Genetics Cold Spring Harbor Laboratory; 1972 pp. 431–433
    [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][PubMed]
    [Google Scholar]
  25. Amit-Romach E, Sklan D, Uni Z. Microflora ecology of the chicken intestine using 16S ribosomal DNA primers. Poult Sci 2004; 83:1093–1098 [View Article][PubMed]
    [Google Scholar]
  26. Sheng Y, Fan F, Jensen O, Zhong Z, Kan B et al. Dual zinc transporter systems in Vibrio cholerae promote competitive advantages over gut microbiome. Infect Immun 2015; 83:3902–3908 [View Article][PubMed]
    [Google Scholar]
  27. Ott SJ, Musfeldt M, Ullmann U, Hampe J, Schreiber S. Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora. J Clin Microbiol 2004; 42:2566–2572 [View Article][PubMed]
    [Google Scholar]
  28. Iwanaga M, Yamamoto K, Higa N, Ichinose Y, Nakasone N et al. Culture conditions for stimulating cholera toxin production by Vibrio cholerae O1 El Tor. Microbiol Immunol 1986; 30:1075–1083 [View Article][PubMed]
    [Google Scholar]
  29. Ferreyra JA, Wu KJ, Hryckowian AJ, Bouley DM, Weimer BC et al. Gut microbiota-produced succinate promotes C. difficile infection after antibiotic treatment or motility disturbance. Cell Host Microbe 2014; 16:770–777 [View Article][PubMed]
    [Google Scholar]
  30. Curtis MM, Hu Z, Klimko C, Narayanan S, Deberardinis R et al. The gut commensal Bacteroides thetaiotaomicron exacerbates enteric infection through modification of the metabolic landscape. Cell Host Microbe 2014; 16:759–769 [View Article][PubMed]
    [Google Scholar]
  31. Ng KM, 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 [View Article][PubMed]
    [Google Scholar]
  32. Goldie AH, Sanwal BD. Genetic and physiological characterization of Escherichia coli mutants deficient in phosphoenolpyruvate carboxykinase activity. J Bacteriol 1980; 141:1115–1121[PubMed]
    [Google Scholar]
  33. Miranda RL, Conway T, Leatham MP, Chang DE, Norris WE et al. Glycolytic and gluconeogenic growth of Escherichia coli O157:H7 (EDL933) and E. coli K-12 (MG1655) in the mouse intestine. Infect Immun 2004; 72:1666–1676 [View Article][PubMed]
    [Google Scholar]
  34. Liu Z, Wang H, Zhou Z, Naseer N, Xiang F et al. Differential thiol-based switches jump-start Vibrio cholerae pathogenesis. Cell Rep 2016; 14:347–354 [View Article][PubMed]
    [Google Scholar]
  35. Johansson ME, Sjövall H, Hansson GC. The gastrointestinal mucus system in health and disease. Nat Rev Gastroenterol Hepatol 2013; 10:352–361 [View Article][PubMed]
    [Google Scholar]
  36. Robbe C, Capon C, Coddeville B, Michalski JC. Structural diversity and specific distribution of O-glycans in normal human mucins along the intestinal tract. Biochem J 2004; 384:307–316 [View Article][PubMed]
    [Google Scholar]
  37. 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 USA 1999; 96:4586–4591 [View Article][PubMed]
    [Google Scholar]
  38. Chang DE, Smalley DJ, Tucker DL, Leatham MP, Norris WE et al. Carbon nutrition of Escherichia coli in the mouse intestine. Proc Natl Acad Sci USA 2004; 101:7427–7432 [View Article][PubMed]
    [Google Scholar]
  39. Conway T, Krogfelt KA, Cohen PS. The life of commensal Escherichia coli in the mammalian intestine. EcoSal Plus 2004; 1: [View Article][PubMed]
    [Google Scholar]
  40. Freter R, Brickner H, Fekete J, Vickerman MM, Carey KE. Survival and implantation of Escherichia coli in the intestinal tract. Infect Immun 1983; 39:686–703[PubMed]
    [Google Scholar]
  41. Tchawa Yimga M, Leatham MP, Allen JH, Laux DC, Conway T et al. Role of gluconeogenesis and the tricarboxylic acid cycle in the virulence of Salmonella enterica serovar Typhimurium in BALB/c mice. Infect Immun 2006; 74:1130–1140 [View Article][PubMed]
    [Google Scholar]
  42. Alteri CJ, Mobley HL. Escherichia coli physiology and metabolism dictates adaptation to diverse host microenvironments. Curr Opin Microbiol 2012; 15:3–9 [View Article][PubMed]
    [Google Scholar]
  43. Rohmer L, Hocquet D, Miller SI. Are pathogenic bacteria just looking for food? metabolism and microbial pathogenesis. Trends Microbiol 2011; 19:341–348 [View Article][PubMed]
    [Google Scholar]
  44. Waligora EA, Fisher CR, Hanovice NJ, Rodou A, Wyckoff EE et al. Role of intracellular carbon metabolism pathways in Shigella flexneri virulence. Infect Immun 2014; 82:2746–2755 [View Article][PubMed]
    [Google Scholar]
  45. Miller CP, Bohnhoff M. Changes in the mouse's enteric microflora associated with enhanced susceptibility to salmonella infection following streptomycin treatment. J Infect Dis 1963; 113:59–66 [View Article][PubMed]
    [Google Scholar]
  46. Spiga L, Winter MG, Furtado de Carvalho T, Zhu W, Hughes ER et al. An oxidative central metabolism enables Salmonella to utilize microbiota-derived succinate. Cell Host Microbe 2017; 22:291.e6–301.e6 [View Article][PubMed]
    [Google Scholar]
  47. Curtis MM, Hu Z, Klimko C, Narayanan S, Deberardinis R et al. The gut commensal Bacteroides thetaiotaomicron exacerbates enteric infection through modification of the metabolic landscape. Cell Host Microbe 2014; 16:759–769 [View Article][PubMed]
    [Google Scholar]
  48. Lehninger AL, Nelson DL, Cox MM. Lehninger Principles of Biochemistry, 5th ed. New York: W.H. Freeman; 2008
    [Google Scholar]
  49. Yildiz FH, Visick KL. Vibrio biofilms: so much the same yet so different. Trends Microbiol 2009; 17:109–118 [View Article][PubMed]
    [Google Scholar]
  50. Katzianer DS, Wang H, Carey RM, Zhu J. "Quorum non-sensing": social cheating and deception in Vibrio cholerae. Appl Environ Microbiol 2015; 81:3856–3862 [View Article][PubMed]
    [Google Scholar]
  51. Zhu J, Mekalanos JJ. Quorum sensing-dependent biofilms enhance colonization in Vibrio cholerae. Dev Cell 2003; 5:647–656 [View Article][PubMed]
    [Google Scholar]
  52. Chao YP, Patnaik R, Roof WD, Young RF, Liao JC. Control of gluconeogenic growth by pps and pck in Escherichia coli. J Bacteriol 1993; 175:6939–6944 [View Article][PubMed]
    [Google Scholar]
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