1887

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Volume 149, Issue 7

Phosphoenolpyruvate phosphotransferase system and -acetylglucosamine metabolism in Free

Abstract

, a bacterium of biotechnological interest due to its ability to produce mosquitocidal toxins, is unable to use sugars as carbon source. However, genes encoding HPr and EI proteins belonging to a PTS were cloned, sequenced and characterized. Both HPr and EI proteins were fully functional for phosphoenolpyruvate-dependent transphosphorylation in complementation assays using extracts from mutants for one of these proteins. HPr(His) was purified from wild-type and a Ser46/Gln mutant of , and used for phosphorylation experiments using extracts from either or as kinase source. The results showed that both phosphorylated forms, P-Ser46-HPr and P-His15-HPr, could be obtained. The findings also proved indirectly the existence of an HPr kinase activity in . The genetic structure of these genes has some unusual features, as they are co-transcribed with genes encoding metabolic enzymes related to -acetylglucosamine (GlcNAc) catabolism (, and an undetermined ). In fact, this bacterium was able to utilize this amino sugar as carbon and energy source, but a null mutant had lost this characteristic. Investigation of GlcNAc uptake and streptozotocin inhibition in both a wild-type and a null mutant strain led to the proposal that GlcNAc is transported and phosphorylated by an EII element of the PTS, as yet uncharacterized. In addition, GlcNAc-6-phosphate deacetylase and GlcN-6-phosphate deaminase activities were determined; both were induced in the presence of GlcNAc. These results, together with the authors' recent findings of the presence of a phosphofructokinase activity, are strongly indicative of a glycolytic pathway in . They also open new possibilities for genetic improvements in industrial applications.

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Keyword(s): FBP, fructose 1,6-bisphosphate , HPr, histidine-containing phosphocarrier protein of the PTS , HPrK/P, HPr kinase/phosphatase , PEP, phosphoenolpyruvate , PFK, phosphofructokinase , PTS, phosphoenolpyruvate : sugar phosphotransferase system and Stz, streptozotocin
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2003-07-01
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References

  1. Alexander B., Priest F. G. 1990; Numerical classification and identification of Bacillus sphaericus including some strains pathogenic for mosquito larvae. J Gen Microbiol 136:367–376
     [Google Scholar]
  2. Alice A. F. 2001 Sistema de fosfotransferasa dependiente del fosfoenolpiruvato ( PTS ) y represión catabólica en Bacillus subtilis PhD Thesis, University of Buenos Aires;
     [Google Scholar]
  3. Alice A. F., Perez-Martinez G., Sanchez-Rivas C. 2002; Existence of a true phosphofructokinase in Bacillus sphaericus : cloning and sequencing of the pfk gene. Appl Environ Microbiol 68:6410–6415
     [Google Scholar]
  4. Altschul S. F., Madden T. L., Schaeffer A. A., Zhang A., Miller W., Lipman D. J. 1997; Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3398–3402
     [Google Scholar]
  5. Ammer J., Brennenstuhl M., Schindler P., Höltje J. V., Zähner H. 1979; Phosphorylation of streptozotocin during uptake via the phosphoenolpyruvate : sugar phosphotransferase system in Escherichia coli . Antimicrob Agents Chemother 16:801–807
     [Google Scholar]
  6. Bates C. J., Pasternak C. A. 1965; Further studies on the regulation of amino sugar metabolism in Bacillus subtilis . Biochem J 96:147–154
     [Google Scholar]
  7. Baumann P., Clark M. A., Baumann L., Broadwell A. H. 1991; Bacillus sphaericus as a mosquito pathogen: properties of the organism and its toxins. Microbiol Rev 55:425–436
     [Google Scholar]
  8. Charles J. F., Nielsen-Le Roux C., Delécluse A. 1996; Bacillus sphaericus toxins: molecular biology and mode of action. Annu Rev Entomol 41:451–472
     [Google Scholar]
  9. Charrier V., Buckley E., Parsonage D., Galinier A., Darbon E., Jaquinod M., Forest E., Deutscher J., Clairbone A. 1997; Cloning and sequencing of two enterococcal glpK genes and regulation of the encoded glycerol kinases by phosphoenolpyruvate-dependent, phosphotransferase system-catalyzed phosphorylation of a single histidyl residue. J Biol Chem 272:14166–14174
     [Google Scholar]
  10. Clarke J. S., Pasternak C. A. 1962; The regulation of amino sugar metabolism in Bacillus subtilis . Biochem J 84:185–191
     [Google Scholar]
  11. Couch T. L. 2000; Industrial fermentation and formulation of entomopathogenic bacteria. In Entomopathogenic Bacteria: from Laboratory to Field Application pp  297–316 Edited by F J. Charles and others Dordrecht: Kluwer;
     [Google Scholar]
  12. deBarjac H., Veron M., Cosmao Dumanoir V. 1980; Caractérisation biochimique et sérologique de souches de Bacillus sphaericus pathogènes ou non pour les moustiques. Ann Microbiol 131B:191–201
     [Google Scholar]
  13. Deutscher J., Küster E., Bergstedt U., Charrier V., Hillen W. 1995; Protein kinase-dependent HPr/CcpA interaction links glycolytic activity to catabolite repression in Gram-positive bacteria. Mol Microbiol 15:1049–1053
     [Google Scholar]
  14. Dossonnet V., Monedero V., Zagorec M., Galinier A., Perez-Martinez G., Deutscher J. 2000; Phosphorylation of HPr by the bifunctional HPr kinase/P-Ser-HPr phosphatase from Lactobacillus casei controls catabolite repression and inducer exclusion but not inducer expulsion. J Bacteriol 182:2582–2590
     [Google Scholar]
  15. Fujita Y., Miwa Y. Galinier A., Deutscher J. 1995; Specific recognition of the Bacillus subtilis gnt cis -acting catabolite-responsive element by a protein complex formed between CcpA and seryl-phosphorylated HPr. Mol Microbiol 17:953–960
     [Google Scholar]
  16. Galinier A., Kravanja M., Engelmann R., Hengstenberg W., Kilhoffer M. C., Deutscher J., Hiech J. 1998; New protein kinase and protein phosphatase families mediate signal transduction in bacterial catabolite repression. Proc Natl Acad Sci U S A 95:1823–1828
     [Google Scholar]
  17. Gonzy-Tréboul G., Zagorec M., Rain-Guian M. C., Steinmetz M. 1989; Phosphoenolpyruvate : sugar phosphotransferase system of Bacillus subtilis : nucleotide sequence of ptsX , ptsH and the 5′ end of the ptsI and evidence for a ptsHI operon. Mol Microbiol 3:103–112
     [Google Scholar]
  18. Guérout-Fleury A. M., Shazand K., Frandsen N., Stragier P. 1995; Antibiotic-resistance cassettes for Bacillus subtilis . Gene 167:335–336
     [Google Scholar]
  19. Haldenwang W. G. 1995; The sigma factors of Bacillus subtilis . Microbiol Rev 59:1–30
     [Google Scholar]
  20. Hengstenberg W., Penberthy W., Hill K., Morse M. 1969; Phosphotransferase system of Staphylococcus aureus : its requirement for the accumulation and metabolism of galactosides. J Bacteriol 99:383–388
     [Google Scholar]
  21. Henkin T. M., Grundy F. J., Nicholson W. L., Chamblis G. H. 1991; Catabolite repression of α-amylase gene expression in Bacillus subtilis involves a trans-acting gene product homologous to the Escherichia coli LacI and GalR repressors. Mol Microbiol 5:575–584
     [Google Scholar]
  22. Himmelreich R., Hilbert H., Plagens H., Pirkl E., Li B. C., Herrmann R. 1996; Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae . Nucleic Acids Res 24:4420–4449
     [Google Scholar]
  23. Hoischen C., Dijkstra A., Rottem S., Reizer J., Saier M. H. Jr 1993; Presence of protein constituents of the Gram-positive bacterial phosphotransferase regulatory system in Acholeplasma laidlawii . J Bacteriol 175:6599–6604
     [Google Scholar]
  24. Iwamoto R., Imanaga Y. 1991; Direct evidence of the Entner–Doudoroff pathway operating in the metabolism of d-glucosamine in bacteria. J Biochem 109:66–69
     [Google Scholar]
  25. Jacobson G. R., Poy F., Lengeler J. W. 1990; Inhibition of Streptococcus mutans by the antibiotic streptozotocin: mechanisms of uptake and the selection of carbohydrate-negative mutants. Infect Immun 58:543–549
     [Google Scholar]
  26. Jault J. M., Fieulaine S., Nessler S., Gonzalo P., Di Pietro A., Deutscher J., Galinier A. 2000; The HPr kinase from Bacillus subtilis is a homo-oligomeric enzyme which exhibits strong positive cooperativity for nucleotide and fructose 1,6-bisphosphate binding. J Biol Chem 275:1773–1780
     [Google Scholar]
  27. Jones-Mortimer M. C., Kornberg H. L. 1980; Amino-sugar transport systems in Escherichia coli K12. J Gen Microbiol 117:369–376
     [Google Scholar]
  28. Kravanja M., Engelmann R., Dossonnet V. 7 other authors 1999; The hprK gene of Enterococcus faecalis encodes a novel bifunctional enzyme: the HPr kinase/phosphatase. Mol Microbiol 31:59–66
     [Google Scholar]
  29. Krych V. K., Johnson J. L., Yousten A. A. 1980; Deoxyribonucleic acid homologies among strains of Bacillus sphaericus . Int J Syst Bacteriol 30:476–482
     [Google Scholar]
  30. Lai X., Ingram L. O. 1995; Discovery of a ptsHI operon, which includes a third gene ( ptsT ), in the thermophile Bacillus stearothermophilus . Microbiology 141:1443–1449
     [Google Scholar]
  31. Landt O., Grunert H. P., Hanh U. 1990; A general method for rapid site-directed mutagenesis using the polymerase chain reaction. Gene 96:125–128
     [Google Scholar]
  32. Lengeler J. 1980; Analysis of the physiological effects of the antibiotic streptozotocin on Escherichia coli K12 and other sensitive bacteria. Arch Microbiol 128:196–203
     [Google Scholar]
  33. Ludwig H., Homuth G., Schmalisch M., Dyka F. M., Hecker M., Stülke J. 2001; Transcription of glycolytic genes and operons in Bacillus subtilis : evidence for the presence of multiple levels of control of the gapA operon. Mol Microbiol 41:409–422
     [Google Scholar]
  34. Martin-Verstraete I., Débarbouillé M., Klier A., Rapoport G. 1990; Levanase operon of Bacillus subtilis includes a fructose-specific phosphotransferase system regulating the expression of the operon. J Mol Biol 214:657–671
     [Google Scholar]
  35. Mijakovic I., Poncet S., Galinier A. 9 other authors 2002; Pyrophosphate-producing protein dephosphorylation by HPr kinase/phosphorylase: a relic of early life?. Proc Natl Acad Sci U S A 99:13442–13447
     [Google Scholar]
  36. Mitchell W., Reizer J., Herring C., Hoischen C., Saier M. H. Jr 1993; Identification of a phosphoenolpyruvate : fructose phosphotransferase system (fructose-1-phosphate forming) in Listeria monocytogenes . J Bacteriol 175:2758–2761
     [Google Scholar]
  37. Mobley H. T., Doyle R. J., Streips U. N., Langemeier S. O. 1982; Transport and incorporation of N -acetylglucosamine in Bacillus subtilis . J Bacteriol 150:8–15
     [Google Scholar]
  38. Peri K. G., Waygood E. B. 1988; Sequence of cloned Enzyme IINag of the phosphoenolpyruvate :  N -acetylglucosamine phosphotransferase system of Escherichia coli . Biochemistry 27:6054–6061
     [Google Scholar]
  39. Plumbridge J. 1989; Sequence of the nagBACD operon in Escherichia coli K12 and pattern of transcription within the nag regulon. Mol Microbiol 3:506–515
     [Google Scholar]
  40. Plumbridge J. 1995; Co-ordinated regulation of amino sugar biosynthesis and degradation: the NagC repressor acts as both an activator and a repressor for the transcription of the glmUS operon and requires two separated NagC binding sites. EMBO J 15:3958–3965
     [Google Scholar]
  41. Plumbridge J. 2001; Regulation of PTS gene expression by the homologous transcriptional regulators, Mlc and NagC, in Escherichia coli (or how two similar repressors can behave differently). J Mol Microbiol Biotechnol 3:371–380
     [Google Scholar]
  42. Porter A. G., Davidson E. W., Liu J. W. 1993; Mosquitocidal toxins of Bacilli and their genetic manipulation for effective biological control of mosquitoes. Microbiol Rev 57:838–861
     [Google Scholar]
  43. Postma P. W., Lengeler J. W., Jacobson G. R. 1993; Phosphoenolpyruvate : carbohydrate phosphotransferase systems of bacteria. Microbiol Rev 57:543–594
     [Google Scholar]
  44. Priest F. G. 2000; Biodiversity of the entomopathogenic, endospore-forming bacteria. In Entomopathogenic Bacteria: from Laboratory to Field Application pp  1–22 Edited by F J. Charles and others Dordrecht: Kluwer;
     [Google Scholar]
  45. Reizer J., Peterkofsky A., Romano A. H. 1988; Evidence for the presence of heat-stable protein (HPr) and ATP-dependent HPr kinase in heterofermentative lactobacilli lacking phosphoenolpyruvate : glycose phosphotransferase activity. Proc Natl Acad Sci U S A 85:2041–2045
     [Google Scholar]
  46. Reizer J., Hoischen C., Titgemeyer F., Rivolta C., Rabus R., Stülke J., Karamata D., Saier M. H. Jr, Hillen W. 1998; A novel protein kinase that controls carbon catabolite repression in bacteria. Mol Microbiol 27:1157–1169
     [Google Scholar]
  47. Reizer J., Bachem S., Reizer A., Arnaud M., Saier M. H. Jr, Stülke J. 1999a; Novel phosphotransferase system genes revealed by genome analysis – the complete complement of PTS proteins encoded within the genome of Bacillus subtilis . Microbiology 145:3419–3429
     [Google Scholar]
  48. Reizer J., Reizer A., Lagrou M. J., Folger K. R., Kendall Stover C., Saier M. H. Jr 1999b; Novel phosphotransferase systems revealed by bacterial genome analysis: the complete repertoire of pts genes in Pseudomonas aeruginosa . J Mol Microbiol Biotechnol 1:289–293
     [Google Scholar]
  49. Rippere K. E., Johnson J. L., Yousten A. A. 1997; DNA similarities among mosquito-pathogens and nonpathogenic strains of Bacillus sphaericus . Int J Syst Bacteriol 47:214–216
     [Google Scholar]
  50. Romano A., Saier M. H. Jr 1992; Evolution of bacterial phosphoenolpyruvate : sugar phosphotransferase system. I. Physiological and organismic considerations. In The Evolution of Metabolic Function pp  143–170 Edited by Mortlock R. P. Boca Raton, FL: CRC Press;
     [Google Scholar]
  51. Roossien F. F., Brink J., Robillard G. T. 1983; A simple procedure for the synthesis of [32P]phosphoenolpyruvate via the pyruvate kinase exchange reaction at equilibrium. Biochim Biophys Acta 760:185–187
     [Google Scholar]
  52. Russell B. L., Jelley S. A., Yousten A. A. 1989; Carbohydrate metabolism in the mosquito pathogen Bacillus sphaericus 2362. Appl Environ Microbiol 55:294–297
     [Google Scholar]
  53. Saier M. H. Jr, Chavaux S., Cook G. M., Deutscher J., Paulsen I. T., Reizer J., Ye J. J. 1996; Catabolite repression and inducer control in Gram-positive bacteria. Microbiology 142:217–230
     [Google Scholar]
  54. Sambrook J., Fritsh E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual Cold Spring Harbor NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  55. Steinhauer K., Jepp T., Hillen W., Stülke J. 2002; A novel mode of control of Mycoplasma pneumoniae HPr kinase/phosphatase activity reflects its parasitic lifestyle. Microbiology 148:3277–3284
     [Google Scholar]
  56. Stülke J., Hillen W. 1999; Carbon catabolite repression in bacteria. Curr Opin Microbiol 2:195–201
     [Google Scholar]
  57. Stülke J., Martin-Verstraete I., Zagorec M., Rose M., Klier A., Rapoport G. 1997; Induction of the Bacillus subtilis ptsGHI operon by glucose is controlled by a novel antiterminator, GlcT. Mol Microbiol 25:65–78
     [Google Scholar]
  58. Stülke J., Arnaud M., Rapoport G., Martin-Verstraete I. 1998; PRD – a protein domain involved in PTS-dependent induction and carbon catabolite repression of catabolic operons in bacteria. Mol Microbiol 28:865–874
     [Google Scholar]
  59. Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G. 1997; The CLUSTAL X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882
     [Google Scholar]
  60. Titgemeyer F., Hillen W. 2002; Global control of sugar metabolism: a gram-positive solution. Antonie Van Leeuwenhoek 82:59–71
     [Google Scholar]
  61. Titgemeyer F., Walkenhorst J., Reizer J., Stuiver M., Cui X., Saier M. H. Jr 1995; Identification and characterization of phosphoenolpyruvate : fructose phosphotransferase in three Streptomyces species. Microbiology 141:51–58
     [Google Scholar]
  62. Vogler A. P., Lengeler J. W. 1989; Analysis of the nag regulon from Escherichia coli K12 and Klebsiella pneumoniae and of its regulation. Mol Gen Genet 219:97–105
     [Google Scholar]
  63. Wagner A., Küster-Schöck E., Hillen W. 2000; Sugar uptake and carbon catabolite repression in Bacillus megaterium strains with inactivated ptsHI . J Mol Microbiol Biotechnol 2:587–592
     [Google Scholar]
  64. Wang F., Xiao X., Saito A., Schrempf H. 2002; Streptomyces olivaceoviridis possesses a phosphotransferase system that mediates specific, phosphoenolpyruvate-dependent uptake of N -acetylglucosamine. Mol Genet Genomics 268:344–351
     [Google Scholar]
  65. Zhu P. P., Herzberg O., Peterkofsky A. 1998; Topography of the interaction of HPr(Ser) kinase with HPr. Biochemistry 37:11762–11770
     [Google Scholar]
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Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
Microbiology 149, 1687 (2003); https://doi.org/10.1099/mic.0.26231-0
/content/journal/micro/10.1099/mic.0.26231-0
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