1887

Abstract

The phosphoenolpyruvate-dependent phosphotransferase system (PTS) is widely found among Gram-positive bacteria. It is the major source of carbohydrate transport in the dental pathogen . The transported carbohydrates are fermented to produce large amounts of lactic acid which initiates dental caries. The authors have isolated the gene for the mannitol-specific Enzyme II (EII) component of the PTS, , and the adjacent gene, which is located in the same operon. The gene is located between and the genes and . The nucleotide sequence of the and loci has been determined. The deduced gene product of consists of 589 amino acids with a molecular mass of 620 kDa. It exhibits similarity with the gene products from other organisms. However, the similarity between these proteins is generally restricted to the 470 amino-terminal residues of the protein. This region would correspond to the EIICB domains of the PTS. The authors have previously shown that the gene product exhibits 766% similarity to the carboxyl-terminal 143 amino acids of the product and that the gene encodes the EIIA domain of the PTS. Thus, the genes that encode the EIICB and the EIIA domains are separated by approximately 2250 bp. In many organisms, all of the EII domains may be fused together to form one molecule. The fact that these domains are separated by this distance in supports the hypothesis that various functional domains of the PTS have been rearranged during evolution. The sequence of the 119 carboxyl-terminal amino acids of the gene product also displays homology to the carboxyl-terminal end of the EIIB domain of various mannitol PTSs. Thus, this domain may have been duplicated in during evolution of the operon. The gene is located in the same operon structure as but these loci are separated by an intragenic space. The precise 5′ end of the locus cannot be determined either by transcription–translation assays or based upon nucleotide sequence analysis because of the apparent lack of a ribosome-binding site preceding the gene. The deduced gene product, which consists of approximately 650 amino acids with a molecular mass of 753 kDa, exhibits limited similarity to several potential transcriptional regulators. However, the exact function of this locus is currently unknown.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-7-1565
2000-07-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/7/1461565a.html?itemId=/content/journal/micro/10.1099/00221287-146-7-1565&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
    [Google Scholar]
  2. Boyd D. A., Cvitkovitch D. G., Hamilton I. R. 1994; Sequence and expression of the genes for HPr (ptsH) and enzyme I (ptsI) of the phosphoenolpyruvate-dependent phosphotransferase transport system from Streptococcus mutans. Infect Immun 62:1156–1165
    [Google Scholar]
  3. Bullock W. O., Fernandez J. M., Short J. M. 1987; XL1-Blue: a high efficiency plasmid transforming recA Escherichia coli strain with beta-galactosidase selection. Biotechniques 5:376–378
    [Google Scholar]
  4. Chen E. Y., Seeberg P. H. 1985; Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA 4:165–170 [CrossRef]
    [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395 [CrossRef]
    [Google Scholar]
  6. Dodd I. B., Egan J. B. 1990; Improved detection of helix–turn–helix DNA-binding motifs in protein sequences. Nucleic Acids Res 18:5019–5026 [CrossRef]
    [Google Scholar]
  7. Drucker D. B., Melville T. H. 1968; Fermentation end-products of cariogenic and non-cariogenic streptococci. Arch Oral Biol 13:565–570 [CrossRef]
    [Google Scholar]
  8. Fischer R., Hengstenberg W. 1992; Mannitol-specific Enzyme II of the phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus carnosus. Sequence and expression in Escherichia coli and structural comparison with the Enzyme IImannitol of Escherichia coli. Eur J Biochem 204:963–969 [CrossRef]
    [Google Scholar]
  9. Fischer R., Eisermann R., Reiche B., Hengstenberg W. 1989; Cloning, sequencing and overexpression of the mannitol-specific Enzyme-III-encoding gene of Staphylococcus carnosus. Gene 82:249–257 [CrossRef]
    [Google Scholar]
  10. Fischer R., von Strandmann R. P., Hengstenberg W. 1991; Mannitol-specific phosphoenolpyruvate-dependent phosphotransferase system of Enterococcus faecalis: molecular cloning and nucleotide sequences of the Enzyme IIIMtl gene and the mannitol-1-phosphate dehydrogenase gene, expression in Escherichia coli, and comparison of the gene products with similar enzymes. J Bacteriol 173:3709–3715
    [Google Scholar]
  11. Henikoff S. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359 [CrossRef]
    [Google Scholar]
  12. Henstra S. A., Tolner B., ten Hoeve Duurkens R. H., Konings W. N., Robillard G. T. 1996; Cloning, expression, and isolation of the mannitol transport protein from the thermophilic bacterium Bacillus stearothermophilus. J Bacteriol 178:5586–5591
    [Google Scholar]
  13. Henstra S. A., Tuinhof M., Duurkens R. H., Robillard G. T. 1999; The Bacillus stearothermophilus mannitol regulator, MtlR, of the phosphotransferase system. A DNA-binding protein, regulated by HPr and IICBmtl-dependent phosphorylation. J Biol Chem 274:4754–4763 [CrossRef]
    [Google Scholar]
  14. Hiratsuka K., Wang B., Sato Y., Kuramitsu H. 1998; Regulation of sucrose-6-phosphate hydrolase activity in Streptococcus mutans: characterization of the scrR gene. Infect Immun 66:3736–3743
    [Google Scholar]
  15. Honeyman A. L., Curtiss R. III 1992; Isolation, characterization, and nucleotide sequence of the Streptococcus mutans mannitol-phosphate dehydrogenase gene and the mannitol-specific factor III gene of the phosphoenolpyruvate phosphotransferase system. Infect Immun 60:3369–3375
    [Google Scholar]
  16. Honeyman A. L., Curtiss R. III 1993; Isolation, characterization, and nucleotide sequence of the Streptococcus mutans lactose-specific enzyme II (lacE) gene of the PTS and the phospho-β-galactosidase (lacG) gene. J Gen Microbiol 139:2685–2694 [CrossRef]
    [Google Scholar]
  17. Kunst F., Ogasawara N., Moszer I.148 other authors 1997; The complete genome sequence of the gram-positive bacterium Bacillus subtilis. Nature 390:249–256 [CrossRef]
    [Google Scholar]
  18. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
    [Google Scholar]
  19. Lee C. A., Saier M. H. Jr 1983; Mannitol-specific Enzyme II of the bacterial phosphotransferase system. III. The nucleotide sequence of the permease gene. J Biol Chem 258:10761–10767
    [Google Scholar]
  20. Macrina F. L., Evans R. P., Tobian J. A., Hartley D. L., Clewell D. B., Jones K. R. 1983; Novel shuttle plasmid vehicles for Escherichia–Streptococcus transgeneric cloning. Gene 25:145–150 [CrossRef]
    [Google Scholar]
  21. Manoil C., Beckwith J. 1985; TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A 82:8129–8133 [CrossRef]
    [Google Scholar]
  22. Moran C. P. Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. 1982; Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet 186:339–346 [CrossRef]
    [Google Scholar]
  23. Murchison H. H., Barrett J. F., Cardineau G. A., Curtiss R. III 1986; Transformation of Streptococcus mutans with chromosomal and shuttle plasmid (pYA629) DNAs. Infect Immun 54:273–282
    [Google Scholar]
  24. Nag D. K., Huang H. V., Berg D. E. 1988; Bidirectional chain-termination nucleotide sequencing: transposon Tn5seq1 as a mobile source of primer sites. Gene 64:135–145 [CrossRef]
    [Google Scholar]
  25. Perez-Casal J., Caparon M. G., Scott J. R. 1991; Mry, a trans-acting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems. J Bacteriol 173:2617–2624
    [Google Scholar]
  26. Postma P. W., Lengeler J. W. 1985; Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. Microbiol Rev 49:232–269
    [Google Scholar]
  27. Reiche B., Frank R., Deutscher J., Meyer N., Hengstenberg W. 1988; Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: purification and characterization of the mannitol-specific Enzyme IIImtl of Staphylococcus aureus and Staphylococcus carnosus and homology with the Enzyme IImtl of Escherichia coli. Biochem 27:6512–6516 [CrossRef]
    [Google Scholar]
  28. van de Rijn I., Kessler R. E. 1980; Growth characteristics of group A streptococci in a new chemically defined medium. Infect Immun 27:444–448
    [Google Scholar]
  29. Rosey E. L., Stewart G. C. 1992; Nucleotide and deduced amino acid sequences of the lacR, lacABCD, and lacFE genes encoding the repressor, tagatose 6-phosphate gene cluster, and sugar-specific phosphotransferase system components of the lactose operon of Streptococcus mutans. J Bacteriol 174:6159–6170
    [Google Scholar]
  30. 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]
  31. Saier M. H. Jr, Yamada M., Erni B.7 other authors 1988; Sugar permeases of the bacterial phosphoenolpyruvate-dependent phosphotransferase system: sequence comparisons. FASEB J 2:199–208
    [Google Scholar]
  32. Sambrook J., Fritsch E. F., Maniatis T. 1989 In Molecular Cloning: a Laboratory Manual, 2nd edn. pp. 140–141 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  33. Sato Y., Yamamoto Y., Suzuki R., Kizaki H., Kuramitsu H. K. 1991; Construction of scrA::lacZ gene fusions to investigate regulation of the sucrose PTS of Streptococcus mutans. FEMS Microbiol Lett 63:339–345
    [Google Scholar]
  34. Schachtele C. F., Mayo J. A. 1973; Phosphoenolpyruvate-dependent glucose transport in oral streptococci. J Dental Res 52:1209–1215 [CrossRef]
    [Google Scholar]
  35. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517 [CrossRef]
    [Google Scholar]
  36. Tabor S., Richardson C. C. 1987; DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A 84:4767–4771 [CrossRef]
    [Google Scholar]
  37. Takami H., Nakasone K., Ogasawara N.7 other authors 1999; Sequencing of three lambda clones from the genome of alkaliphilic Bacillus sp. strain C-125. Extremophiles 3:29–34 [CrossRef]
    [Google Scholar]
  38. Vellanoweth R. L., Rabinowitz J. C. 1992; The influence of ribosome-binding-site elements on translational efficiency in Bacillus subtilis and Escherichia coli in vivo. Mol Microbiol 6:1105–1114 [CrossRef]
    [Google Scholar]
  39. Yamane K., Kumano M., Kurita K. 1996; The 25°–36° region of the Bacillus subtilis chromosome: determination of the sequence of a 146 kb segment and identification of 113 genes. Microbiology 142:3047–3056 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-7-1565
Loading
/content/journal/micro/10.1099/00221287-146-7-1565
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