1887

Abstract

The phosphoenopyruvate:carbohydrate phosphotransferase system (PTS) enables – and other bacteria – to recognize and transport exogenous carbon sources for energy, including the six-carbon sugar alcohol, mannitol. The mannitol-specific PTS transporter is encoded by and its expression is expected to be regulated by the putative repressor encoded by the gene. Here, we show that overexpression inhibits growth in medium supplied with mannitol as the sole carbon source and represses MtlA-mediated biofilm formation. We demonstrate that when is grown in non-mannitol medium, knocking out leads to both increased MtlA protein and mRNA levels, with these increases being especially pronounced in non-glucose sugars. We propose that in non-mannitol, non-glucose growth conditions, MtlR is a major regulator of transcription. Surprisingly, with regard to expression, transcript and protein levels are highest in mannitol medium, conditions where expression should not be repressed. We further show that MtlR levels increase during growth of the bacteria and linger in cells switched from mannitol to non-mannitol medium. Our data suggests an expression paradigm for where MtlR acts as a transcriptional repressor responsible for calibrating MtlA levels during environmental transitions.

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2017-12-01
2024-03-29
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References

  1. Ali M, Lopez AL, You YA, Kim YE, Sah B et al. The global burden of cholera. Bull World Health Organ 2012; 90:209–218 [View Article][PubMed]
    [Google Scholar]
  2. Colwell RR. Infectious disease and environment: cholera as a paradigm for waterborne disease. Int Microbiol 2004; 7:285–289[PubMed]
    [Google Scholar]
  3. 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]
  4. Lutz C, Erken M, Noorian P, Sun S, McDougald D. Environmental reservoirs and mechanisms of persistence of Vibrio cholerae. Front Microbiol 2013; 4:375 [View Article][PubMed]
    [Google Scholar]
  5. Merrell DS, Hava DL, Camilli A. Identification of novel factors involved in colonization and acid tolerance of Vibrio cholerae. Mol Microbiol 2002; 43:1471–1491 [View Article][PubMed]
    [Google Scholar]
  6. Moorthy S, Watnick PI. Genetic evidence that the Vibrio cholerae monolayer is a distinct stage in biofilm development. Mol Microbiol 2004; 52:573–587 [View Article][PubMed]
    [Google Scholar]
  7. Schild S, Tamayo R, Nelson EJ, Qadri F, Calderwood SB et al. Genes induced late in infection increase fitness of Vibrio cholerae after release into the environment. Cell Host Microbe 2007; 2:264–277 [View Article][PubMed]
    [Google Scholar]
  8. Xu Q, Dziejman M, Mekalanos JJ. Determination of the transcriptome of Vibrio cholerae during intraintestinal growth and midexponential phase in vitro. Proc Natl Acad Sci USA 2003; 100:1286–1291 [View Article][PubMed]
    [Google Scholar]
  9. Deutscher J, Aké FM, Derkaoui M, Zébré AC, Cao TN et al. The bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system: regulation by protein phosphorylation and phosphorylation-dependent protein-protein interactions. Microbiol Mol Biol Rev 2014; 78:231–256 [View Article][PubMed]
    [Google Scholar]
  10. Houot L, Chang S, Pickering BS, Absalon C, Watnick PI. The phosphoenolpyruvate phosphotransferase system regulates Vibrio cholerae biofilm formation through multiple independent pathways. J Bacteriol 2010; 192:3055–3067 [View Article][PubMed]
    [Google Scholar]
  11. Houot L, Watnick PI. A novel role for enzyme I of the Vibrio cholerae phosphoenolpyruvate phosphotransferase system in regulation of growth in a biofilm. J Bacteriol 2008; 190:311–320 [View Article][PubMed]
    [Google Scholar]
  12. Moorthy S, Watnick PI. Identification of novel stage-specific genetic requirements through whole genome transcription profiling of Vibrio cholerae biofilm development. Mol Microbiol 2005; 57:1623–1635 [View Article][PubMed]
    [Google Scholar]
  13. Deutscher J, Francke C, Postma PW. How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria. Microbiol Mol Biol Rev 2006; 70:939–1031 [View Article][PubMed]
    [Google Scholar]
  14. Postma PW, Lengeler JW, Jacobson GR. Phosphoenolpyruvate:carbohydrate phosphotransferase systems of bacteria. Microbiol Rev 1993; 57:543–594[PubMed]
    [Google Scholar]
  15. Houot L, Chang S, Absalon C, Watnick PI. Vibrio cholerae phosphoenolpyruvate phosphotransferase system control of carbohydrate transport, biofilm formation, and colonization of the germfree mouse intestine. Infect Immun 2010; 78:1482–1494 [View Article][PubMed]
    [Google Scholar]
  16. Groisillier A, Shao Z, Michel G, Goulitquer S, Bonin P et al. Mannitol metabolism in brown algae involves a new phosphatase family. J Exp Bot 2014; 65:559–570 [View Article][PubMed]
    [Google Scholar]
  17. Iwamoto K, Shiraiwa Y. Salt-regulated mannitol metabolism in algae. Mar Biotechnol 2005; 7:407–415 [View Article][PubMed]
    [Google Scholar]
  18. Oddo E, Saiano F, Alonzo G, Bellini E. An investigation of the seasonal pattern of mannitol content in deciduous and evergreen species of the Oleaceae growing in northern Sicily. Ann Bot 2002; 90:239–243 [View Article][PubMed]
    [Google Scholar]
  19. Rousvoal S, Groisillier A, Dittami SM, Michel G, Boyen C et al. Mannitol-1-phosphate dehydrogenase activity in Ectocarpus siliculosus, a key role for mannitol synthesis in brown algae. Planta 2011; 233:261–273 [View Article][PubMed]
    [Google Scholar]
  20. Rambhatla P, Kumar S, Floyd JT, Varela MF. Molecular cloning and characterization of mannitol-1-phosphate dehydrogenase from Vibrio cholerae. J Microbiol Biotechnol 2011; 21:914–920 [View Article][PubMed]
    [Google Scholar]
  21. Joyet P, Derkaoui M, Bouraoui H, Deutscher J. PTS-mediated regulation of the transcription activator MtlR from different species: surprising differences despite strong sequence conservation. J Mol Microbiol Biotechnol 2015; 25:94–105 [View Article][PubMed]
    [Google Scholar]
  22. Watanabe S, Hamano M, Kakeshita H, Bunai K, Tojo S et al. Mannitol-1-phosphate dehydrogenase (MtlD) is required for mannitol and glucitol assimilation in Bacillus subtilis: possible cooperation of mtl and gut operons. J Bacteriol 2003; 185:4816–4824 [View Article][PubMed]
    [Google Scholar]
  23. Kumar S, Smith KP, Floyd JL, Varela MF. Cloning and molecular analysis of a mannitol operon of phosphoenolpyruvate-dependent phosphotransferase (PTS) type from Vibrio cholerae O395. Arch Microbiol 2011; 193:201–208 [View Article][PubMed]
    [Google Scholar]
  24. Liu JM, Livny J, Lawrence MS, Kimball MD, Waldor MK et al. Experimental discovery of sRNAs in Vibrio cholerae by direct cloning, 5S/tRNA depletion and parallel sequencing. Nucleic Acids Res 2009; 37:e46 [View Article][PubMed]
    [Google Scholar]
  25. Ymele-Leki P, Houot L, Watnick PI. Mannitol and the mannitol-specific enzyme IIB subunit activate Vibrio cholerae biofilm formation. Appl Environ Microbiol 2013; 79:4675–4683 [View Article][PubMed]
    [Google Scholar]
  26. Zhou YY, Zhang HZ, Liang WL, Zhang LJ, Zhu J et al. Plasticity of regulation of mannitol phosphotransferase system operon by CRP-cAMP complex in Vibrio cholerae. Biomed Environ Sci 2013; 26:831–840 [View Article][PubMed]
    [Google Scholar]
  27. Chang H, Replogle JM, Vather N, Tsao-Wu M, Mistry R et al. A cis-regulatory antisense RNA represses translation in Vibrio cholerae through extensive complementarity and proximity to the target locus. RNA Biol 2015; 12:136–148 [View Article][PubMed]
    [Google Scholar]
  28. Mustachio LM, Aksit S, Mistry RH, Scheffler R, Yamada A et al. The Vibrio cholerae mannitol transporter is regulated posttranscriptionally by the MtlS small regulatory RNA. J Bacteriol 2012; 194:598–606 [View Article][PubMed]
    [Google Scholar]
  29. Figge RM, Ramseier TM, Saier MH. The mannitol repressor (MtlR) of Escherichia coli. J Bacteriol 1994; 176:840–847 [View Article][PubMed]
    [Google Scholar]
  30. Wang HY, Yan MY, Zhao YW, Kan B. [Transcriptional repressor gene - - mtlR of mannitol PTS operon in Vibrio cholerae]. Wei Sheng Wu Xue Bao 2007; 47:522–525[PubMed]
    [Google Scholar]
  31. Tan K, Clancy S, Borovilos M, Zhou M, Hörer S et al. The mannitol operon repressor MtlR belongs to a new class of transcription regulators in bacteria. J Biol Chem 2009; 284:36670–36679 [View Article][PubMed]
    [Google Scholar]
  32. Herrington DA, Hall RH, Losonsky G, Mekalanos JJ, Taylor RK et al. Toxin, toxin-coregulated pili, and the toxR regulon are essential for Vibrio cholerae pathogenesis in humans. J Exp Med 1988; 168:1487–1492 [View Article][PubMed]
    [Google Scholar]
  33. Donnenberg MS, Kaper JB. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun 1991; 59:4310–4317[PubMed]
    [Google Scholar]
  34. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 2009; 6:343–345 [View Article][PubMed]
    [Google Scholar]
  35. Lee SH, Angelichio MJ, Mekalanos JJ, Camilli A. Nucleotide sequence and spatiotemporal expression of the Vibrio cholerae vieSAB genes during infection. J Bacteriol 1998; 180:2298–2305[PubMed]
    [Google Scholar]
  36. Papenfort K, Förstner KU, Cong JP, Sharma CM, Bassler BL. Differential RNA-seq of Vibrio cholerae identifies the VqmR small RNA as a regulator of biofilm formation. Proc Natl Acad Sci USA 2015; 112:E766E775 [View Article][PubMed]
    [Google Scholar]
  37. Zhang D, Manos J, Ma X, Belas R, Karaolis DK. Transcriptional analysis and operon structure of the tagA-orf2-orf3-mop-tagD region on the Vibrio pathogenicity island in epidemic V. cholerae. FEMS Microbiol Lett 2004; 235:199–207 [View Article][PubMed]
    [Google Scholar]
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