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Volume 148, Issue 3

The gene encoding the novel general stress protein Gsp62 Free

Abstract

The general stress protein Gsp62 was purified using two-dimensional gel electrophoresis and its 25 N-terminal amino acid sequence determined. Analysis of the corresponding gene revealed that the product is a 172 aa protein. Transcriptional analysis of gave evidence for a monocistronic mRNA, the synthesis of which was induced at the onset of stationary phase and in response to heat shock, acid pH, detergents (i.e. SDS or bile salts), ethanol, tert-butyl hydroperoxide, sodium chloride and, to a lesser extent, hydrogen peroxide. 5′ rapid amplification of cDNA ends by PCR experiments showed that transcription initiates 30 nt upstream of the ATG start codon. Although expression was induced in response to various stresses, its disruption had no significant effect on the cell survival after each individual stress. Two-dimensional protein gels from wild-type and mutant cells revealed no pleiotropic effect of the mutation on protein synthesis. Transcriptional fusions with the β-galactosidase genes showed that an inverted repeat located upstream of the promoter is required for transcriptional induction by environmental stresses but not by entrance into stationary phase. Two distinct mechanisms responding to different signals are thus involved in induction.

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Keyword(s): Enterococcus faecalis , general stress protein , Gsp and tBOOH, tert-butyl hydroperoxide; 2-D, two-dimensional; IR, inverted repeat; RACE, rapid amplification of cDNA ends
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2002-03-01
2024-04-19
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References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [CrossRef]
     [Google Scholar]
  2. Bardonnet N., Blanco C. 1992; ‘uidA -antibiotic-resistance cassettes for insertion mutagenesis, gene fusions and genetic constructions. FEMS Microbiol Lett 93:243–247
     [Google Scholar]
  3. Becker L. A., Cetin M. S., Hutkins R. W., Benson A. K. 1998; Identification of a gene encoding the alternative sigma factor σB from Listeria monocytogenes and its role in osmotolerance. J Bacteriol 180:4547–4554
     [Google Scholar]
  4. Bolotin A., Manger S., Malarme K., Ehrlich S. D., Sorokin A. 1999; Low-redundancy sequencing of the entire Lactococcus lactis IL1403 genome. Antonie Leeuwenhoek 76:27–76 [CrossRef]
     [Google Scholar]
  5. Boutibonnes P., Giard J. C., Hartke A., Thammavongs B., Auffray Y. 1993; Characterization of the heat shock response in Enterococcus faecalis. . Antonie Leeuwenhoek 64:47–55 [CrossRef]
     [Google Scholar]
  6. Dower J. W., Miller J. F., Ragsdale C. W. 1988; High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145 [CrossRef]
     [Google Scholar]
  7. Flahaut S., Benachour A., Giard J. C., Boutibonnes P., Auffray Y. 1996a; Defense against lethal treatments and de novo protein synthesis induced by NaCl in Enterococcus faecalis ATCC 19433. Arch Microbiol 165:317–324 [CrossRef]
     [Google Scholar]
  8. Flahaut S., Boutibonnes P., Auffray Y., Frère J. 1996b; Comparison of the bile salts and sodium dodecyl sulfate stress responses in Enterococcus faecalis. . Appl Environ Microbiol 62:2416–2420
     [Google Scholar]
  9. Flahaut S., Hartke A., Giard J. C., Benachour A., Boutibonnes P., Auffray Y. 1996c; Relationship between stress response toward bile salts, acid and heat treatment in Enterococcus faecalis . FEMS Microbiol Lett 138:49–54 [CrossRef]
     [Google Scholar]
  10. Flahaut S., Boutibonnes P., Auffray Y. 1997a; Les Enterocoques dans l’environnement proche de l’homme. Can J Microbiol 43:699–708 [CrossRef]
     [Google Scholar]
  11. Flahaut S., Boutibonnes P., Auffray Y., Frère J. 1997b; Relationship between the thermotolerance and the increase of DnaK and GroEL synthesis in Enterococcus faecalis ATCC19433. J Basic Microbiol 37:251–258 [CrossRef]
     [Google Scholar]
  12. Flahaut S., Hartke A., Giard J. C., Auffray Y. 1997c; Alkaline stress response in Enterococcus faecalis : adaptation, cross-protection, and changes in protein synthesis. Appl Environ Microbiol 63:812–814
     [Google Scholar]
  13. Flahaut S., Laplace J. M., Auffray Y., Frère J. 1998; The oxidative stress response in Enterococcus faecalis : relationship between H2O2 tolerance and H2O2 stress proteins. Lett Appl Microbiol 26:259–264 [CrossRef]
     [Google Scholar]
  14. Frère J., Benachour A., Novel M., Novel G. 1993; Identification of the theta-type minimal replicon of the Lactococcus lactis subsp. lactis CNRZ270 lactose protease plasmid pUCL22. Curr Microbiol 27:97–102 [CrossRef]
     [Google Scholar]
  15. Gaillot O., Pellegrini E., Bregenholt S., Nair S., Berche P. 2000; The ClpP serine protease is essential for the intracellular parasitism and virulence of Listeria monocytogenes . Mol Microbiol 35:1286–1294
     [Google Scholar]
  16. Giard J. C., Hartke A., Flahaut S., Benachour A., Boutibonnes P., Auffray Y. 1997; Glucose starvation response in Enterococcus faecalis JH2-2: survival and proteins analysis. Res Microbiol 148:27–35 [CrossRef]
     [Google Scholar]
  17. Giard J. C., Capiaux H., Auffray Y., Hartke A., Rincé A. 2000; Inactivation of the stress- and starvation-inducible gls24 operon has a pleiotropic effect on cell morphology, stress sensitivity and gene expression in Enterococcus faecalis . J Bacteriol 182:4512–4520 [CrossRef]
     [Google Scholar]
  18. Giard J. C., Laplace J. M., Pichereau V., Benachour A., Leboeuf C., Flahaut S., Auffray Y., Hartke A., Rincé A. 2001; The stress proteome of Enterococcus faecalis . Electrophoresis 22:2947–2954 [CrossRef]
     [Google Scholar]
  19. Hecker M., Völker U. 1998; Non-specific general and multiple stress resistance of growth-restricted Bacillus subtilis cells by the expression of the σB regulon. Mol Microbiol 29:1129–1136 [CrossRef]
     [Google Scholar]
  20. Hecker M., Schumann W., Völker U. 1996; Heat-shock and general stress response in Bacillus subtilis. Mol Microbiol 19. 417–428 [CrossRef]
  21. Holo H., Nes I. F. 1995; Transformation of Lactococcus by electroporation. Methods Mol Biol 47:195–199
     [Google Scholar]
  22. Israelsen H., Madsen S. M., Vrang A., Hansen E. B., Johansen E. 1995; Cloning and partial characterization of regulated promoters from Lactococcus lactis Tn 917 - lacZ integrants with the new promoter probe vector, pAK80. Appl Environ Microbiol 61:2540–2547
     [Google Scholar]
  23. Jett B. D., Huycke M. M., Gilmore M. S. 1994; Virulence of enterococci. Clin Microbiol Rev 7:462–478
     [Google Scholar]
  24. Jones D., Shattock P. M. 1960; The location of the group antigen of group D Streptococcus . J Gen Microbiol 23:335–343 [CrossRef]
     [Google Scholar]
  25. Laplace J. M., Boutibonnes P., Auffray Y. 1996; Unusual resistance and acquired tolerance to cadmium chloride in Enterococcus faecalis. J Basic Microbiol 36:311–317 [CrossRef]
     [Google Scholar]
  26. Miller J. M. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  27. Mundt J. O. 1986; Enterococci. In Bergey’s Manual of Systematic Bacteriology pp 1063–1065 Edited by Sneath P. H. A. Mair N. S., Sharpe M. E., Holt J. G. Baltimore: Williams & Wilkins;
     [Google Scholar]
  28. Rincé A., Flahaut S., Auffray Y. 2000; Identification of general stress genes in Enterococcus faecalis . Int J Food Microbiol 55:87–91 [CrossRef]
     [Google Scholar]
  29. Rincé A., Giard J. C., Pichereau V., Flahaut S., Auffray Y. 2001; Identification and characterization of gsp65 , an organic hydroperoxide resistance ( ohr ) gene encoding a general stress protein in Enterococcus faecalis . J Bacteriol 183:1482–1488 [CrossRef]
     [Google Scholar]
  30. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  31. Terzaghi B. E., Sandine W. E. 1975; Improved medium for lactic streptococci and their bacteriophages. Appl Environ Microbiol 29:807–813
     [Google Scholar]
  32. Wiedmann M., Arvik T. J., Boor K. J. 1998; General stress transcription factor σB and its role in acid tolerance and virulence of Listeria monocytogenes . J Bacteriol 180:3650–3656
     [Google Scholar]
  33. Wu S., Tomasz A., de Lencastre H. 1996; Sigma-B, a putative operon encoding alternate sigma factor of Staphylococcus aureus RNA polymerase: molecular cloning and DNA sequencing. J Bacteriol 178:6036–6042
     [Google Scholar]
  34. Yagi Y., Clewell D. B. 1980; Recombination-deficient mutant of Streptococcus faecalis . J Bacteriol 143:966–970
     [Google Scholar]
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The Enterococcus faecalis gene encoding the novel general stress protein Gsp62
Microbiology 148, 703 (2002); https://doi.org/10.1099/00221287-148-3-703
/content/journal/micro/10.1099/00221287-148-3-703
/content/journal/micro/10.1099/00221287-148-3-703
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