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. 1997 Jun;63(6):2111–2116. doi: 10.1128/aem.63.6.2111-2116.1997

Molecular cloning and analysis of the ptsHI operon in Lactobacillus sake.

R Stentz 1, R Lauret 1, S D Ehrlich 1, F Morel-Deville 1, M Zagorec 1
PMCID: PMC168499  PMID: 9172326

Abstract

The ptsH and ptsI genes of Lactobacillus sake, encoding the general enzymes of the phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS), were cloned and sequenced. HPr (88 amino acids), encoded by ptsH, and enzyme I (574 amino acids), encoded by ptsI, are homologous to the corresponding known enzymes of other bacteria. Nucleotide sequence and mRNA analysis showed that the two genes are cotranscribed in a large transcript encoding both HPr and enzyme I. The transcription of ptsHI was shown to be independent of the carbon source. Four ptsI mutants were constructed by single-crossover recombination. For all mutants, growth on PTS carbohydrates was abolished. Surprisingly, the growth rates of mutants on ribose and arabinose, two carbohydrates which are not transported by the PTS, were accelerated. This unexpected phenotype suggests that the PTS negatively controls ribose and arabinose utilization in L. sake by a mechanism different from the regulation involving HPr described for other gram-positive bacteria.

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Selected References

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  1. Anba J., Bidnenko E., Hillier A., Ehrlich D., Chopin M. C. Characterization of the lactococcal abiD1 gene coding for phage abortive infection. J Bacteriol. 1995 Jul;177(13):3818–3823. doi: 10.1128/jb.177.13.3818-3823.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson D. G., McKay L. L. Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol. 1983 Sep;46(3):549–552. doi: 10.1128/aem.46.3.549-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Reuse H., Danchin A. The ptsH, ptsI, and crr genes of the Escherichia coli phosphoenolpyruvate-dependent phosphotransferase system: a complex operon with several modes of transcription. J Bacteriol. 1988 Sep;170(9):3827–3837. doi: 10.1128/jb.170.9.3827-3837.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Reuse H., Roy A., Danchin A. Analysis of the ptsH-ptsI-crr region in Escherichia coli K-12: nucleotide sequence of the ptsH gene. Gene. 1985;35(1-2):199–207. doi: 10.1016/0378-1119(85)90172-6. [DOI] [PubMed] [Google Scholar]
  6. Deutscher J., Reizer J., Fischer C., Galinier A., Saier M. H., Jr, Steinmetz M. Loss of protein kinase-catalyzed phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system, by mutation of the ptsH gene confers catabolite repression resistance to several catabolic genes of Bacillus subtilis. J Bacteriol. 1994 Jun;176(11):3336–3344. doi: 10.1128/jb.176.11.3336-3344.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deutscher J., Sauerwald H. Stimulation of dihydroxyacetone and glycerol kinase activity in Streptococcus faecalis by phosphoenolpyruvate-dependent phosphorylation catalyzed by enzyme I and HPr of the phosphotransferase system. J Bacteriol. 1986 Jun;166(3):829–836. doi: 10.1128/jb.166.3.829-836.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fujita Y., Miwa Y., Galinier A., Deutscher J. 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. 1995 Sep;17(5):953–960. doi: 10.1111/j.1365-2958.1995.mmi_17050953.x. [DOI] [PubMed] [Google Scholar]
  10. Gay P., Cordier P., Marquet M., Delobbe A. Carbohydrate metabolism and transport in Bacillus subtilis. A study of ctr mutations. Mol Gen Genet. 1973 Mar 19;121(4):355–368. doi: 10.1007/BF00433234. [DOI] [PubMed] [Google Scholar]
  11. Gonzy-Tréboul G., Karmazyn-Campelli C., Stragier P. Developmental regulation of transcription of the Bacillus subtilis ftsAZ operon. J Mol Biol. 1992 Apr 20;224(4):967–979. doi: 10.1016/0022-2836(92)90463-t. [DOI] [PubMed] [Google Scholar]
  12. Gonzy-Tréboul G., Zagorec M., Rain-Guion M. C., Steinmetz M. Phosphoenolpyruvate:sugar phosphotransferase system of Bacillus subtilis: nucleotide sequence of ptsX, ptsH and the 5'-end of ptsI and evidence for a ptsHI operon. Mol Microbiol. 1989 Jan;3(1):103–112. doi: 10.1111/j.1365-2958.1989.tb00109.x. [DOI] [PubMed] [Google Scholar]
  13. Gonzy-Tréboul G., de Waard J. H., Zagorec M., Postma P. W. The glucose permease of the phosphotransferase system of Bacillus subtilis: evidence for IIGlc and IIIGlc domains. Mol Microbiol. 1991 May;5(5):1241–1249. doi: 10.1111/j.1365-2958.1991.tb01898.x. [DOI] [PubMed] [Google Scholar]
  14. Kohlbrecher D., Eisermann R., Hengstenberg W. Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: molecular cloning and nucleotide sequence of the Staphylococcus carnosus ptsI gene and expression and complementation studies of the gene product. J Bacteriol. 1992 Apr;174(7):2208–2214. doi: 10.1128/jb.174.7.2208-2214.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lauret R., Morel-Deville F., Berthier F., Champomier-Verges M., Postma P., Ehrlich S. D., Zagorec M. Carbohydrate Utilization in Lactobacillus sake. Appl Environ Microbiol. 1996 Jun;62(6):1922–1927. doi: 10.1128/aem.62.6.1922-1927.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leloup L., Ehrlich S. D., Zagorec M., Morel-Deville F. Single-crossover integration in the Lactobacillus sake chromosome and insertional inactivation of the ptsI and lacL genes. Appl Environ Microbiol. 1997 Jun;63(6):2117–2123. doi: 10.1128/aem.63.6.2117-2123.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lopilato J., Bortner S., Beckwith J. Mutations in a new chromosomal gene of Escherichia coli K-12, pcnB, reduce plasmid copy number of pBR322 and its derivatives. Mol Gen Genet. 1986 Nov;205(2):285–290. doi: 10.1007/BF00430440. [DOI] [PubMed] [Google Scholar]
  18. Odermatt A., Solioz M. Two trans-acting metalloregulatory proteins controlling expression of the copper-ATPases of Enterococcus hirae. J Biol Chem. 1995 Mar 3;270(9):4349–4354. doi: 10.1074/jbc.270.9.4349. [DOI] [PubMed] [Google Scholar]
  19. Odermatt A., Suter H., Krapf R., Solioz M. Primary structure of two P-type ATPases involved in copper homeostasis in Enterococcus hirae. J Biol Chem. 1993 Jun 15;268(17):12775–12779. [PubMed] [Google Scholar]
  20. Postma P. W., Lengeler J. W., Jacobson G. R. Phosphoenolpyruvate:carbohydrate phosphotransferase systems of bacteria. Microbiol Rev. 1993 Sep;57(3):543–594. doi: 10.1128/mr.57.3.543-594.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reizer J., Hoischen C., Reizer A., Pham T. N., Saier M. H., Jr Sequence analyses and evolutionary relationships among the energy-coupling proteins Enzyme I and HPr of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. Protein Sci. 1993 Apr;2(4):506–521. doi: 10.1002/pro.5560020403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reizer J., Saier M. H., Jr, Deutscher J., Grenier F., Thompson J., Hengstenberg W. The phosphoenolpyruvate:sugar phosphotransferase system in gram-positive bacteria: properties, mechanism, and regulation. Crit Rev Microbiol. 1988;15(4):297–338. doi: 10.3109/10408418809104461. [DOI] [PubMed] [Google Scholar]
  23. Saffen D. W., Presper K. A., Doering T. L., Roseman S. Sugar transport by the bacterial phosphotransferase system. Molecular cloning and structural analysis of the Escherichia coli ptsH, ptsI, and crr genes. J Biol Chem. 1987 Nov 25;262(33):16241–16253. [PubMed] [Google Scholar]
  24. Saier M. H., Jr, Chauvaux S., Cook G. M., Deutscher J., Paulsen I. T., Reizer J., Ye J. J. Catabolite repression and inducer control in Gram-positive bacteria. Microbiology. 1996 Feb;142(Pt 2):217–230. doi: 10.1099/13500872-142-2-217. [DOI] [PubMed] [Google Scholar]
  25. Schnierow B. J., Yamada M., Saier M. H., Jr Partial nucleotide sequence of the pts operon in Salmonella typhimurium: comparative analyses in five bacterial genera. Mol Microbiol. 1989 Jan;3(1):113–118. doi: 10.1111/j.1365-2958.1989.tb00110.x. [DOI] [PubMed] [Google Scholar]
  26. Squires C., Squires C. L. The Clp proteins: proteolysis regulators or molecular chaperones? J Bacteriol. 1992 Feb;174(4):1081–1085. doi: 10.1128/jb.174.4.1081-1085.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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