1887

Abstract

The discovery of a operon in XL-65-6 coupled with our previous report of a operon (Lai Ingram, 175, 6441-6450, 1993) demonstrates that this thermophilic organism contains all of the genes required for cellobiose uptake by the phosphoenolpyruvate-dependent phosphotransferase system (PTS). Genes encoding the two general PTS proteins, HPr () and enzyme l (), were cloned and sequenced. These form an operon which includes a third small gene () of unknown function (encoded product 18428). Both and were expressed at high levels from a single plasmid in and complemented corresponding host mutations. Although the translated sequences for these genes were similar to homologues from Gram-positive mesophiles (64-77% identity), the gene products were unusual in having a higher predicted pi and fewer negatively charged amino acid residues. Enzyme I also contained more alanine and leucine than mesophilic counterparts. Interestingly, inhibited the growth of mutants at 37°C. No such inhibition was observed during incubation at a lower temperature (30°C) or in DH5, which is wild-type for . The predicted translation product from contained a high proportion of basic amino acids (27%) and had a high predicted pl (pH 117), properties similar to bacterial histone-like proteins, but did not exhibit homology to any sequences in the current database. Regions upstream and downstream from the operon contain genes with homology to and (wall-associated protein), respectively.

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1995-06-01
2021-05-18
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References

  1. Benson D., Lipman D.J., Ostell J. 1993; GenBank.. Nucleic Acids Res 21:2963–2965
    [Google Scholar]
  2. Chen Y., Fairbrother W.J., Wright P.E. 1993; Threedimensional structures of the central regulatory proteins of the bacterial phosphotransferase system, HPr and Enzyme IIAglc.. J Cell Biochem 51:75–82
    [Google Scholar]
  3. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Res 12:387–395
    [Google Scholar]
  4. Drlica K., Rouviere-Yaniv J. 1987; Histonelike proteins of bacteria.. Microbiol Rev 51:301–319
    [Google Scholar]
  5. Foster S.J. 1993; Molecular analysis of three major wall-associated proteins of Bacillus subtilis 168: evidence for processing of the product of a gene encoding a 258 kDa precursor, two-domain ligand-binding protein.. Mol Microbiol 8:299–310
    [Google Scholar]
  6. Fox C.F., Wilson G. 1968; The role of a phosphoenolpyruvate-dependent kinase system in beta-glucoside catabolism in Escherichia coli.. Proc Natl Acad Sci USA 59988–995
    [Google Scholar]
  7. Fraenkel D.G., Falcoz-Kelly F., Horecker B.L. 1964; The utilization of glucose 6-phosphate by glucokinaseless and wild-type strains of Escherichia coli.. Proc Natl Acad Sci USA 521207–1213
    [Google Scholar]
  8. Gonzy-Tréboul G., Zagorec M., Rain-Guion M.-C., Steinmetz M. 1989; Phosphoenolpyruvate: sugar phosphotransferase system of Bacillus subtilis: nucleotide sequence of ptsX,ptsH and the 5̓-end of ptsI and evidence for a ptsHI operon.. Mol Microbiol 3:103–112
    [Google Scholar]
  9. Lai X., Ingram L.O. 1993; Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearo-thermophilus XL-65-6 and functional expression in Escherichia coli.. J Bacteriol 175:6441–6450
    [Google Scholar]
  10. Lévy S., De Reuse H., Danchin A. 1989; Antisense expression at the ptsH-ptsI locus of Escherichia coli.. FEMS Microbiol Lett 57:35–38
    [Google Scholar]
  11. Miller C.G. 1987; Protein degradation and proteolytic modification.. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp. 680–691 Neidhardt F.C., Ingraham J.L., Low K.B., Magasanik B., Schaechter M., Umbarger H.E. Edited by Washington, DC:: American Society for Microbiology.;
    [Google Scholar]
  12. Moran C.P. Jr 1993; RNA polymerase and transcription factors.. In Bacillus subtilis and Other Gram-Positive Bacteria: Biochemistry, Physiology, and Molecular Genetics pp. 653–667 Sonenshein A.L., Hoch J.A., Losick R. Edited by Washington, DC:: American Society for Microbiology.;
    [Google Scholar]
  13. Mozhaev V.V., Martinek K. 1984; Structure-stability relationships in proteins: new approaches to stabilizing enzymes.. Enzyme Microb Technol 6:50–59
    [Google Scholar]
  14. Murphy N., McConnell D.J., Cantwell B.A. 1984; The DNA sequence of the gene and genetic control sites for the excreted Bacillus subtilis enzyme β-glucanase.. Nucleic Acids Res 12:5355–5367
    [Google Scholar]
  15. Nochur S.V., Jacobson G.R., Roberts M.F., Demain A.L. 1992; Mode of sugar phosphorylation in Clostridium thermocellum.. Appl Biochem Biotechnol 33:33–41
    [Google Scholar]
  16. Parker L.L., Hall B.G. 1990; Characterization and nucleotide sequence of the cryptic cel operon of Escherichia coli K-12.. Genetics 124:455–471
    [Google Scholar]
  17. Patni N.J., Alexander J.K. 1971; Catabolism of fructose and mannitol in Clostridium thermocellum: presence of phosphoenolpyruvate: fructose phosphotransferase, fructose 1-phosphate kinase, phosphoenolpyruvate: mannitol phosphotransferase, and mannitol 1-phosphate dehydrogenase in cell extracts.. J Bacteriol 105:226–231
    [Google Scholar]
  18. Postma P.W., Lengeler J.W., Jacobson G.R. 1993; Phos-phoenolpyruvate : carbohydrate phosphotransferase system of bacteria.. Microbiol Rev 57:543–594
    [Google Scholar]
  19. Reizer J., Hoischen C., Reizer A., Saier M.H. 1993; Sequence analysis and evolutionary relationships among the energy coupling proteins enzyme I and HPr of the bacterial phosphoenolpyruvate : sugar phosphotransferase system.. Protein Sci 2:506–521
    [Google Scholar]
  20. Saier M.H. Jr Reizer J. 1992; Proposed uniform nomenclature for the proteins and protein domains of the bacterial phosphoenolpyruvate: sugar phosphotransferase system.. J Bacteriol 174:1433–1438
    [Google Scholar]
  21. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Eaboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  22. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA 745463–5467
    [Google Scholar]
  23. Schägger H., von Jagow G. 1987; Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.. Anal Biochem 166:368–379
    [Google Scholar]
  24. Titgemeyer F. 1986 Genetische Untersuchungen ɀum Sucrose-Stoffmchsel bei Klebsiella pneumoniae. Diploma thesis Universität Osnabrück, Germany.:
    [Google Scholar]
  25. Van Charldorp R., Van Kimmenade A.M., Van Knippenberg P.H. 1981; Sequence and secondary structure of the colicin fragment of Bacillus stearothermophilus 16S ribosomal RNA.. Nucleic Acids Res 9:4909–4917
    [Google Scholar]
  26. Zagorec M., Postma P.W. 1992; Cloning and nucleotide sequence of the ptsG gene of Bacillus subtilis.. Mol & Gen Genet 234:325–328
    [Google Scholar]
  27. Zhu P.-P., Reizer J., Reizer A., Peterkofsky A. 1993; Unique monocistronic operon (ptsH) in Mycoplasma capricolum encoding the phosphocarrier protein, HPr, of the phosphoenolpyruvate: sugar phosphotransferase system.. J Biol Chem 268:26531–26540
    [Google Scholar]
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