1887

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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-3-703
2002-03-01
2020-08-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/3/1480703a.html?itemId=/content/journal/micro/10.1099/00221287-148-3-703&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J.. 1990; Basic local alignment search tool. J Mol Biol215:403–410[CrossRef]
    [Google Scholar]
  2. Bardonnet N., Blanco C.. 1992; ‘uidA -antibiotic-resistance cassettes for insertion mutagenesis, gene fusions and genetic constructions. FEMS Microbiol Lett93:243–247
    [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 Bacteriol180:4547–4554
    [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 Leeuwenhoek76:27–76[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 Leeuwenhoek64:47–55[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 Res16:6127–6145[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 Microbiol165:317–324[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 Microbiol62:2416–2420
    [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 Lett138:49–54[CrossRef]
    [Google Scholar]
  10. Flahaut S., Boutibonnes P., Auffray Y.. 1997a; Les Enterocoques dans l’environnement proche de l’homme. Can J Microbiol43:699–708[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 Microbiol37:251–258[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 Microbiol63:812–814
    [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 Microbiol26:259–264[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 Microbiol27:97–102[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 Microbiol35:1286–1294
    [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 Microbiol148:27–35[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 Bacteriol182:4512–4520[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 . Electrophoresis22:2947–2954[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 Microbiol29:1129–1136[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 Biol47:195–199
    [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 Microbiol61:2540–2547
    [Google Scholar]
  23. Jett B. D., Huycke M. M., Gilmore M. S.. 1994; Virulence of enterococci. Clin Microbiol Rev7:462–478
    [Google Scholar]
  24. Jones D., Shattock P. M.. 1960; The location of the group antigen of group D Streptococcus . J Gen Microbiol23:335–343[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 Microbiol36:311–317[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 pp1063–1065 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 Microbiol55:87–91[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 Bacteriol183:1482–1488[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 Microbiol29:807–813
    [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 Bacteriol180:3650–3656
    [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 Bacteriol178:6036–6042
    [Google Scholar]
  34. Yagi Y., Clewell D. B.. 1980; Recombination-deficient mutant of Streptococcus faecalis . J Bacteriol143:966–970
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-3-703
Loading
/content/journal/micro/10.1099/00221287-148-3-703
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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