1887

Abstract

Δ Δ Δ glucose strains that lack the glucose phosphotransferase system (PTS) and the mannose PTS as well as glucokinase have been widely used by researchers studying the PTS. In this study we show that both fast- and slow-growing spontaneous glucose revertants can be readily obtained from Δ Δ Δ glucose strains. All of the fast-growing revertants either altered the -acetylglucosamine PTS or caused its overproduction by inactivating the NagC repressor protein, which regulates the -acetylglucosamine PTS, and these revertants could utilize either glucose or -acetylglucosamine as a sole carbon source. When a Δ deletion, which abolishes the -acetylglucosamine PTS, was introduced into the Δ Δ Δ glucose strains, fast-growing revertants could no longer be isolated. Based on our results and other studies, it is clear that the -acetylglucosamine PTS is the most easily adaptable PTS for transporting and phosphorylating glucose, other than the glucose PTS and mannose PTS, which are the primary glucose transport systems. While the slow-growing glucoserevertants were not characterized, they were likely mutations that other researchers have observed before and affect other PTSs or sugar kinases.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000596
2018-02-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/2/163.html?itemId=/content/journal/micro/10.1099/mic.0.000596&mimeType=html&fmt=ahah

References

  1. Guan L, Kaback HR. Glucose/sugar transport in bacteria. In Lennarz WJ, Lane MD. (editors) Encyclopedia of Biological Chemistry, 2nd ed. Oxford: Elsevier; 2013 pp. 387–390 [Crossref]
    [Google Scholar]
  2. Curtis SJ, Epstein W. Phosphorylation of d-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase. J Bacteriol 1975; 122:1189–1199[PubMed]
    [Google Scholar]
  3. Hunter IS, Kornberg HL. Glucose transport of Escherichia coli growing in glucose-limited continuous culture. Biochem J 1979; 178:97–101 [View Article][PubMed]
    [Google Scholar]
  4. Postma PW, Lengeler JW, Jacobson GR. Phosphoenolpyruvate:carbohydrate phosphotransferase systems. In Neidhardt FC. (editor) Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd ed. Washington, DC: ASM Press; 1996 pp. 1149–1174
    [Google Scholar]
  5. Tchieu JH, Norris V, Edwards JS, Saier MH. The complete phosphotransferase system in Escherichia coli . J Mol Microbiol Biotechnol 2001; 3:329–346[PubMed]
    [Google Scholar]
  6. Lengeler JW. PTS 50: past, present and future, or diauxie revisited. J Mol Microbiol Biotechnol 2015; 25:79–93 [View Article][PubMed]
    [Google Scholar]
  7. Plumbridge JA. Sequence of the nagBACD operon in Escherichia coli K12 and pattern of transcription within the nag regulon. Mol Microbiol 1989; 3:505–515 [View Article][PubMed]
    [Google Scholar]
  8. Plumbridge J, Kolb A. DNA loop formation between Nag repressor molecules bound to its two operator sites is necessary for repression of the nag regulon of Escherichia coli in vivo . Mol Microbiol 1993; 10:973–981 [View Article][PubMed]
    [Google Scholar]
  9. Bertani G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli . J Bacteriol 1951; 62:293–300[PubMed]
    [Google Scholar]
  10. Miller JH. Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1972
    [Google Scholar]
  11. Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2006; 2:1–11 [View Article][PubMed]
    [Google Scholar]
  12. Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 2000; 97:6640–6645 [View Article][PubMed]
    [Google Scholar]
  13. 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]
  14. Yu D, Ellis HM, Lee EC, Jenkins NA, Copeland NG et al. An efficient recombination system for chromosome engineering in Escherichia coli . Proc Natl Acad Sci USA 2000; 97:5978–5983 [View Article][PubMed]
    [Google Scholar]
  15. Singer M, Baker TA, Schnitzler G, Deischel SM, Goel M et al. A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli . Microbiol Rev 1989; 53:1–24[PubMed]
    [Google Scholar]
  16. Guyer MS, Reed RR, Steitz JA, Low KB. Identification of a sex-factor-affinity site in E. coli as gamma delta. Cold Spring Harb Symp Quant Biol 1981; 45:135–140 [View Article][PubMed]
    [Google Scholar]
  17. Casadaban MJ. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol 1976; 104:541–555 [View Article][PubMed]
    [Google Scholar]
  18. Nichols BP, Shafiq O, Meiners V. Sequence analysis of Tn10 insertion sites in a collection of Escherichia coli strains used for genetic mapping and strain construction. J Bacteriol 1998; 180:6408–6411[PubMed]
    [Google Scholar]
  19. Pennetier C, Domínguez-Ramírez L, Plumbridge J. Different regions of Mlc and NagC, homologous transcriptional repressors controlling expression of the glucose and N-acetylglucosamine phosphotransferase systems in Escherichia coli, are required for inducer signal recognition. Mol Microbiol 2008; 67:364–377 [View Article][PubMed]
    [Google Scholar]
  20. Begley GS, Warner KA, Arents JC, Postma PW, Jacobson GR. Isolation and characterization of a mutation that alters the substrate specificity of the Escherichia coli glucose permease. J Bacteriol 1996; 178:940–942 [View Article][PubMed]
    [Google Scholar]
  21. Oh H, Park Y, Park C. A mutated PtsG, the glucose transporter, allows uptake of d-ribose. J Biol Chem 1999; 274:14006–14011 [View Article][PubMed]
    [Google Scholar]
  22. Notley-Mcrobb L, Ferenci T. Substrate specificity and signal transduction pathways in the glucose-specific enzyme II (EII(Glc)) component of the Escherichia coli phosphotransferase system. J Bacteriol 2000; 182:4437–4442 [View Article][PubMed]
    [Google Scholar]
  23. Zeppenfeld T, Larisch C, Lengeler JW, Jahreis K. Glucose transporter mutants of Escherichia coli K-12 with changes in substrate recognition of IICB(Glc) and induction behavior of the ptsG gene. J Bacteriol 2000; 182:4443–4452 [View Article][PubMed]
    [Google Scholar]
  24. Cao Y, Jin X, Levin EJ, Huang H, Zong Y et al. Crystal structure of a phosphorylation-coupled saccharide transporter. Nature 2011; 473:50–54 [View Article][PubMed]
    [Google Scholar]
  25. Mccoy JG, Levin EJ, Zhou M. Structural insight into the PTS sugar transporter EIIC. Biochim Biophys Acta 2015; 1850:577–585 [View Article][PubMed]
    [Google Scholar]
  26. Raberg M, Peplinski K, Heiss S, Ehrenreich A, Voigt B et al. Proteomic and transcriptomic elucidation of the mutant ralstonia eutropha G+1 with regard to glucose utilization. Appl Environ Microbiol 2011; 77:2058–2070 [View Article][PubMed]
    [Google Scholar]
  27. Orita I, Iwazawa R, Nakamura S, Fukui T. Identification of mutation points in Cupriavidus necator NCIMB 11599 and genetic reconstitution of glucose-utilization ability in wild strain H16 for polyhydroxyalkanoate production. J Biosci Bioeng 2012; 113:63–69 [View Article][PubMed]
    [Google Scholar]
  28. Schnetz K, Sutrina SL, Saier MH, Rak B. Identification of catalytic residues in the beta-glucoside permease of Escherichia coli by site-specific mutagenesis and demonstration of interdomain cross-reactivity between the β-glucoside and glucose systems. J Biol Chem 1990; 265:13464–13471[PubMed]
    [Google Scholar]
  29. Reidl J, Boos W. The malX malY operon of Escherichia coli encodes a novel enzyme II of the phosphotransferase system recognizing glucose and maltose and an enzyme abolishing the endogenous induction of the maltose system. J Bacteriol 1991; 173:4862–4876 [View Article][PubMed]
    [Google Scholar]
  30. Hummel U, Nuoffer C, Zanolari B, Erni B. A functional protein hybrid between the glucose transporter and the N-acetylglucosamine transporter of Escherichia coli . Protein Sci 1992; 1:356–362 [View Article][PubMed]
    [Google Scholar]
  31. Steinsiek S, Bettenbrock K. Glucose transport in Escherichia coli mutant strains with defects in sugar transport systems. J Bacteriol 2012; 194:5897–5908 [View Article][PubMed]
    [Google Scholar]
  32. Miller BG, Raines RT. Identifying latent enzyme activities: substrate ambiguity within modern bacterial sugar kinases. Biochemistry 2004; 43:6387–6392 [View Article][PubMed]
    [Google Scholar]
  33. Miller BG, Raines RT. Reconstitution of a defunct glycolytic pathway via recruitment of ambiguous sugar kinases. Biochemistry 2005; 44:10776–10783 [View Article][PubMed]
    [Google Scholar]
  34. Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V et al. The complete genome sequence of Escherichia coli K-12. Science 1997; 277:1453–1462 [View Article][PubMed]
    [Google Scholar]
  35. Schiefner A, Gerber K, Seitz S, Welte W, Diederichs K et al. The crystal structure of Mlc, a global regulator of sugar metabolism in Escherichia coli . J Biol Chem 2005; 280:29073–29079 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000596
Loading
/content/journal/micro/10.1099/mic.0.000596
Loading

Data & Media loading...

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