Browse > Article

Differential Regulation of the Genes of the Streptococcus pneumoniae dnaK Operon by Ca++  

Kim, Seung-Whan (Department of Pharmacology, University of Ulsan College of Medicine)
Bae, Yong-Goo (College of Pharmacy, Sungkyunkwan University)
Pyo, Suhk-Neung (College of Pharmacy, Sungkyunkwan University)
Rhee, Dong-Kwon (College of Pharmacy, Sungkyunkwan University)
Publication Information
Molecules and Cells / v.23, no.2, 2007 , pp. 239-245 More about this Journal
Abstract
DnaK is a major antigen in Streptococcus pneumoniae, and is induced by a minor shift in temperature (30 to $37^{\circ}C$) but not by ethanol shock. Although HrcA in the presence of $Ca^{{+}{+}}$ represses the expression of both groEL and hrcA, the control of transcription of the dnaK operon is not completely understood. In this study, the dnaK operon of S. pneumoniae (5' hrcA-grpE-dnaK-dnaJ) was cloned and analyzed. It contains large intergenic regions in grpE/dnaK and dnaK/dnaJ. Pulse labeling with [$^{35}S$]-methionine and immunoblot analyses revealed the presence of higher levels of DnaK than of HrcA even in the presence of $Ca^{{+}{+}}$ after heat shock suggesting that $Ca^{{+}{+}}$ differentially regulates the heat shock responses of hrcA and dnaK. By blocking de novo mRNA synthesis with rifampin it was shown that neither the hrcA nor the groEL transcripts were stabilized by heat shock even though dnaK transcripts were stabilized. We conclude that S. pneumoniae uses fine regulation of the transcription of the individual genes of the tetracistronic dnaK operon to cope with the various stresses experienced during infections.
Keywords
Calcium; HrcA; dnaK Operon; Streptococcus pneumoniae;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
Times Cited By Web Of Science : 2  (Related Records In Web of Science)
연도 인용수 순위
1 Choi, I. H, Shim, J. H., Kim, S. W., Kim, S. N., Pyo, S. N., et al. (1999) Limited stress response in Streptococcus pneumoniae. Microbiol. Immunol. 43, 807−812
2 Dalton, T. L. and Scott, J. R. (2004) CovS inactivates CovR and is required for growth under conditions of general stress in Streptococcus pyogenes. J. Bacteriol. 186, 3928−3937
3 Ferretti, J. J., McShan, W. M., Ajdic, D., Savic, D. J., Savic, G., et al. (2001) Complete genome sequence of an M1 strain of Streptococcus pyogenes. Proc. Natl. Acad. Sci. USA 98, 4658− 4663
4 Hamel, J., Martin, D., and Brodeur, B. B. (1997) Heat shock response of Streptococcus pneumoniae: identification of immunoreactive stress proteins. Microb. Pathog. 23, 11−21
5 Tettelin, H., Nelson, K. E., Paulsen, I. T., Eisen, J. A., Read, T. D., et al. (2001) Complete genome sequence of a virulent isolate of Streptococcus pneumoniae. Science 293, 498−506. J. Bacteriol. 175, 1637−1644
6 Kim, S. N., Kim, S. W., Pyo, S. N., and Rhee, D. K. (2001) Molecular cloning and characterization of groESL operon in Streptococcus pneumoniae. Mol. Cells 11, 360−368
7 Narberhaus, F. (1999) Negative regulation of bacterial heat shock genes. Mol. Microbiol. 31, 1−8
8 Jayaraman, G. C., Penders, J. E., and Burne, R. A. (1997) Transcriptional analysis of the Streptococcus mutans hrcA, grpE and dnaK genes and regulation of expression in response to heat shock and environmental acidification. Mol. Microbiol. 25, 329−341
9 Kim, S. W., Choi, I. H., Kim, S. N., Kim, Y. H., Pyo, S. N., et al. (1998) Cloning, expression, and characterization of dnaK in Streptococcus pneumoniae. FEMS Microbiol. Lett. 161, 217− 224
10 Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680−685
11 Sabelnikov, A. G., Greenberg, B., and Lacks, S. A. (1995) An extended −10 promoter alone directs transcription of the DpnII operon of Streptococcus pneumoniae. J. Mol. Biol. 250, 144−155
12 Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
13 Hitomi, M., Nishimura, H., Tsujimoto, Y., Matsui, H., and Watanabe, K. (2003) Identification of a helix-turn-helix motif of Bacillus thermoglucosidasius HrcA essential for binding to the CIRCE element and thermostability of the HrcA-CIRCE complex, indicating a role as a thermosensor. J. Bacteriol. 185, 381−385
14 Paton, J. C., Andrew, P. W., Boulnois, G. J., and Mitchell, T. J. (1993) Molecular analysis of the pathogenicity of Streptococcus pneumoniae: the role of pneumococcal proteins. Annu. Rev. Microbiol. 47, 89−115
15 Homuth, G., Masuda, S., Mogk, A., Kobayashi, Y., and Schumann, W. (1997) The dnaK operon of Bacillus subtilis is heptacistronic. J. Bacteriol. 179, 1153−1164
16 Kwon, H. Y., Kim, S. W., Choi, M. H., Ogunniyi, A. D., Paton, J. C., et al. (2003) Effect of heat shock and mutations in ClpL and ClpP on virulence gene expression in Streptococcus pneumoniae. Infect. Immun. 71, 3757−3765
17 Morimoto, R. I., Tissieres, A., and Georgopoulos, C. (1994) The Biology of Heat Shock Proteins and Molecular Chaperones, Cold Spring Harbor Laboratory Press, Cold Spring Harbor
18 Hoskins, J., Alborn, W. E. Jr, Arnold, J., Blaszczak, L. C., Burgett, S., et al. (2001) Genome of the bacterium Streptococcus pneumoniae strain R6. J. Bacteriol. 183, 5709−5717
19 Homuth, G., Mogk, A., and Schumann, W. (1999) Posttranscriptional regulation of the Bacillus subtilis dnaK operon. Mol. Microbiol. 32, 1183−1197
20 Kwon, H. Y., Kim, S. N., Pyo, S. N., and Rhee, D. K. (2005) $Ca^{2+}$-dependent expression of the CIRCE regulon in Streptococcus pneumoniae. Mol. Microbiol. 55, 456−468
21 Schulz, A. and Schumann, W. (1996) hrcA, the first gene of the Bacillus subtilis dnaK operon encodes a negative regulator of class I heat shock genes. J. Bacteriol. 178, 1088−1093
22 Neidhardt, F. C. and VanBogelen, R. A. (1987) Heat shock response; in E. coli and Salmonella typhimurium: Cellular and Molecular Biology, Neidhardt, F. C., Ingraham, J. L., Low, K. B., Magasanik, B., Schaechter, M., and Umbarger, H. E. (eds.), pp. 1334−1345, ASM Press, Washington, D.C
23 Mogk, A., Homuth, G., Scholz, C., Kim, L., Schmid, F. X., et al. (1997) The GroE chaperonin machine is a major modulator of the CIRCE heat shock regulon of Bacillus subtilis. EMBO J. 16, 4579−4590
24 Rhee, D. K. and Morrison, D. A. (1988) Genetic transformation in Streptococcus pneumoniae: Molecular cloning and characterization of recP, a gene required for genetic recombination. J. Bacteriol. 170, 630−637
25 Lemos, J. A., Chen, Y. Y., and Burne, R. A. (2001) Genetic and physiologic analysis of the groE operon and role of the HrcA repressor in stress gene regulation and acid tolerance in Streptococcus mutans. J. Bacteriol. 183, 6074−6084
26 Gottesman, S., Wickner, S., and Maurizi, M. R. (1997) Protein quality control: triage by chaperones and proteases. Genes Dev. 11, 815–823