1887

Abstract

The main causes of microbial death after heat exposure are not well understood. Here, it is shown that the heat-shock protein ClpP plays a major role in heat-induced growth arrest in . A mutant lacking the ClpP protease was more sensitive to the inhibitory effects of heat, salt and oxidative stress than the isogenic wild-type strain. During growth arrest, this mutant displayed important modifications of its total protein content, including a decreased level of essential metabolic enzymes such as the alcohol dehydrogenase. Analysis of protein carbonylation demonstrated that the ClpP protease plays a role in preventing accelerated protein oxidation. Higher levels of oxidized DnaK, a key modulator of the heat-shock regulon, were observed in the ClpP mutant and these were increased following heat shock. Accumulation of oxidized/inactivated DnaK might explain why the ClpP mutant was unable to properly synthesize DNA and proteins, and why it exhibited an aberrant cell morphology. Even though ClpP plays a minor role in the virulence of in a murine infection model, the data presented here point to the importance of ClpP in oxidative stress defence in preventing heat-induced cell alterations.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.25783-0
2003-02-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/2/mic149407.html?itemId=/content/journal/micro/10.1099/mic.0.25783-0&mimeType=html&fmt=ahah

References

  1. Biswas I, Gruss A, Ehrlich S. D., Maguin E. 1993; High-efficiency gene inactivation and replacement system for gram-positive bacteria. J Bacteriol 175:3628–3635
    [Google Scholar]
  2. Bochner B. R, Lee P. C, Wilson S. W, Cutler C. W., Ames B. N. 1984; AppppA and related adenylated nucleotides are synthesized as a consequence of oxidation stress. Cell 37:225–232
    [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli . J Mol Biol 41:459–472
    [Google Scholar]
  4. Celli J., Trieu-Cuot P. 1998; Circularization of Tn 916 is required for expression of the transposon-encoded transfer functions: characterization of long tetracycline-inducible transcripts reading through the attachment site. Mol Microbiol 28:103–118
    [Google Scholar]
  5. Chastanet A, Prudhomme M, Claverys J. P., Msadek T. 2001; Regulation of Streptococcus pneumoniae clp genes and their role in competence development and stress survival. J Bacteriol 183:7295–7307
    [Google Scholar]
  6. Cruz-Rodz A., Gilmore M. S. 1990; High efficiency introduction of plasmid DNA into glycine treated Enterococcus faecalis by electroporation. Mol Gen Genet 224:152–154
    [Google Scholar]
  7. Davidson J. F, Whyte B, Bissinger P. H., Schiestl R. H. 1996; Oxidative stress is involved in heat-induced cell death in Saccharomyces cerevisiae . Proc Natl Acad Sci U S A 93:5116–5121
    [Google Scholar]
  8. Derre I, Rapoport G., Msadek T. 1999; CtsR, a novel regulator of stress and heat shock response, controls clp and molecular chaperone gene expression in gram-positive bacteria. Mol Microbiol 31:117–131
    [Google Scholar]
  9. Dukan S., Nystrom T. 1998; Bacterial senescence: stasis results in increased and differential oxidation of cytoplasmic proteins leading to developmental induction of the heat shock regulon. Genes Dev 12:3431–3441
    [Google Scholar]
  10. Dukan S., Nystrom T. 1999; Oxidative stress defense and deterioration of growth-arrested Escherichia coli cells. J Biol Chem 274:26027–26032
    [Google Scholar]
  11. Frehel C., Leduc M. 1987; Cytochemical localization of lipopolysaccharides during peptidoglycan degradation of Escherichia coli cells. J Bacteriol 169:210–217
    [Google Scholar]
  12. Gaillot O, Poyart C, Berche P., Trieu-Cuot P. 1997; Molecular characterization and expression analysis of the superoxide dismutase gene from Streptococcus agalactiae . Gene 204:213–218
    [Google Scholar]
  13. 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]
  14. Gaillot O, Bregenholt S, Jaubert F, Di Santo J. P., Berche P. 2001; Stress-induced ClpP serine protease of Listeria monocytogenes is essential for induction of listeriolysin O-dependent protective immunity. Infect Immun 69:4938–4943
    [Google Scholar]
  15. Georgopoulos C, Liberek K, Zylicz M., Ang D. 1994; Properties of the heat shock proteins of Escherichia coli and the autoregulation of the heat shock response. In The Biology of Heat Shock Proteins and Molecular Chaperones pp 209–249 Edited by Morimoto R. I., Tissières A., Georgopoulos C. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  16. Gerth U, Kruger E, Derre I, Msadek T., Hecker M. 1998; Stress induction of the Bacillus subtilis clpP gene encoding a homologue of the proteolytic component of the Clp protease and the involvement of ClpP and ClpX in stress tolerance. Mol Microbiol 28:787–802
    [Google Scholar]
  17. Glaser P, Rusniok C, Buchrieser C. 9 other authors 2002; Genome sequence of Streptococcus agalactiae , a pathogen causing invasive neonatal disease. Mol Microbiol 45:1499–1513
    [Google Scholar]
  18. Gottesman S. 1996; Proteases and their targets in Escherichia coli . Annu Rev Genet 30:465–506
    [Google Scholar]
  19. Gottesman S, Roche E, Zhou Y., Sauer R. T. 1998; The ClpXP and ClpAP proteases degrade proteins with carboxy-terminal peptide tails added by the SsrA-tagging system. Genes Dev 12:1338–1347
    [Google Scholar]
  20. Jenal U., Fuchs T. 1998; An essential protease involved in bacterial cell-cycle control. EMBO J 17:5658–5669
    [Google Scholar]
  21. Jones A. L, Knoll K. M., Rubens C. E. 2000; Identification of Streptococcus agalactiae virulence genes in the neonatal rat sepsis model using signature-tagged mutagenesis. Mol Microbiol 37:1444–1455
    [Google Scholar]
  22. Kim S. W, Choi I. H, Kim S. N, Kim Y. H, Pyo S. N., Rhee D. K. 1998; Molecular cloning, expression, and characterization of dnaK in Streptococcus pneumoniae . FEMS Microbiol Lett 161:217–224
    [Google Scholar]
  23. Lehninger A. L. 1982 Principles of Biochemistry New York: Worth;
    [Google Scholar]
  24. Lodish H, Berk A, Zipursky S. L, Matsudaira P, Baltimore D., Darnell J. E. 2000 Molecular Cell Biology, 4th edn. New York: W. H. Freeman;
    [Google Scholar]
  25. Maurizi M. R, Clark W. P, Kim S. H., Gottesman S. 1990; ClpP represents a unique family of serine proteases. J Biol Chem 265:12546–12552
    [Google Scholar]
  26. Morimoto R. I. 1998; Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes Dev 12:3788–3796
    [Google Scholar]
  27. Morimoto R. I, Tissieres I., Georgopoulos C. 1994; Progress and perspectives on the biology of heat shock proteins and molecular chaperones. In The Biology of Heat Shock Proteins and Molecular Chaperones pp 1–30 Edited by Morimoto R. I., Tissieres I., Georgopoulos C. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Nair S, Frehel C, Nguyen L, Escuyer V., Berche P. 1999; ClpE, a novel member of the HSP100 family, is involved in cell division and virulence of Listeria monocytogenes . Mol Microbiol 31:185–196
    [Google Scholar]
  29. Nair S, Milohanic E., Berche P. 2000a; ClpC ATPase is required for cell adhesion and invasion of Listeria monocytogenes . Infect Immun 68:7061–7068
    [Google Scholar]
  30. Nair S, Derre I, Msadek T, Gaillot O., Berche P. 2000b; CtsR controls class III heat shock gene expression in the human pathogen Listeria monocytogenes . Mol Microbiol 35:800–811
    [Google Scholar]
  31. Newton A. 1972; Role of transcription in the temporary control of development in Caulobacter crescentus . Proc Natl Acad Sci U S A 69:447–451
    [Google Scholar]
  32. Poyart C., Trieu-Cuot P. 1997; A broad-host-range mobilizable shuttle vector for the construction of transcriptional fusions to β-galactosidase in Gram-positive bacteria. FEMS Microbiol Lett 156:193–198
    [Google Scholar]
  33. Poyart C, Lamy M.-C, Boumaila C, Fiedler F., Trieu-Cuot P. 2001a; Regulation of d-alanyl-lipoteichoic acid biosynthesis in Streptococcus agalactiae involves a novel two-component regulatory system. J Bacteriol 183:6324–6334
    [Google Scholar]
  34. Poyart C, Pellegrini E, Gaillot O, Boumaila C, Baptista M., Trieu-Cuot P. 2001b; Contribution of Mn-cofactored superoxide dismutase (SodA) to the virulence of Streptococcus agalactiae . Infect Immun 69:5098–5106
    [Google Scholar]
  35. Poyart-Salmeron C, Trieu-Cuot P, Carlier C, MacGowan A, McLauchlin J., Courvalin P. 1992; Genetic basis of tetracycline resistance in clinical isolates of Listeria monocytogenes . Antimicrob Agents Chemother 36:463–466
    [Google Scholar]
  36. Robertson G. T, Ng W. L, Foley J, Gilmour R., Winkler M. E. 2002; Global transcriptional analysis of clpP mutations of type 2 Streptococcus pneumoniae and their effects on physiology and virulence. J Bacteriol 184:3508–3520
    [Google Scholar]
  37. Rouquette C, de Chastellier C, Nair S., Berche P. 1998; The ClpC ATPase of Listeria monocytogenes is a general stress protein required for virulence and promoting early bacterial escape from the phagosome of macrophages. Mol Microbiol 27:1235–1245
    [Google Scholar]
  38. 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]
  39. Schirmer E. C, Glover J. R, Singer M. A., Lindquist S. 1996; HSP100/Clp proteins: a common mechanism explains diverse functions. Trends Biochem Sci 21:289–296
    [Google Scholar]
  40. Schuchat A. 1998; Epidemiology of group B streptococcal disease in the United States: shifting paradigms. Clinical Microbiol Rev 11:497–513
    [Google Scholar]
  41. Squires C., Squires C. L. 1992; The Clp proteins: proteolysis regulators or molecular chaperones?. J Bacteriol 174:1081–1085
    [Google Scholar]
  42. Stadtman E. R. 1992; Protein oxidation and aging. Science 257:1220–1224
    [Google Scholar]
  43. Tamarit J, Cabiscol E., Ros J. 1998; Identification of the major oxidatively damaged proteins in Escherichia coli cells exposed to oxidative stress. J Biol Chem 273:3027–3032
    [Google Scholar]
  44. Tamura G. S, Kuypers J. M, Smith S, Raff H., Rubens C. E. 1994; Adherence of group B streptococci to cultured epithelial cells: roles of environmental factors and bacterial surface components. Infect Immun 62:2450–2458
    [Google Scholar]
  45. Thomas C., Smith C. 1987; Incompatibility group P plasmids: genetics, evolution, and use in genetic manipulation. Annu Rev Microbiol 41:77–101
    [Google Scholar]
  46. Wang J, Hartling J. A., Flanagan J. M. 1997; The structure of ClpP at 2·3 Å resolution suggests a model for ATP-dependent proteolysis. Cell 91:447–456
    [Google Scholar]
  47. Winzeler E., Shapiro L. 1995; Use of flow cytometry to identify a Caulobacter 4·5S RNA temperature-sensitive mutant defective in the cell cycle. J Mol Biol 251:346–365
    [Google Scholar]
  48. Woo K. M, Chung W. J, Ha D. B, Goldberg A. L., Chung C. H. 1989; Protease Ti from Escherichia coli requires ATP hydrolysis for protean breakdown but not for hydrolysis of small peptides. J Biol Chem 264:2088–2091
    [Google Scholar]
  49. Yanisch-Perron C, Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
  50. Zhou Y, Gottesman S, Hoskins J. R, Maurizi M. R., Wickner S. 2001; The RssB response regulator directly targets σS for degradation by ClpXP. Genes Dev 15:627–637
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.25783-0
Loading
/content/journal/micro/10.1099/mic.0.25783-0
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