This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Yuan, G. Articles by Wong, S. L. Search for Related Content PubMed PubMed Citation Articles by Yuan, G. Articles by Wong, S. L.

 Previous Article  |  Next Article 

J. Bacteriol., Nov 1995, 6462-6468, Vol 177, No. 22
Copyright © 1995, American Society for Microbiology

Isolation and characterization of Bacillus subtilis groE regulatory mutants: evidence for orf39 in the dnaK operon as a repressor gene in regulating the expression of both groE and dnaK

G Yuan and SL Wong
Department of Biological Sciences, University of Calgary, Alberta, Canada.

An inverted repeat sequence known as CIRCE (controlling inverted repeat of chaperone expression) in the Bacillus subtilis groE operon has been suggested to function as an operator. To identify the regulatory gene directly or indirectly involved in CIRCE-mediated heat-inducible groE expression, B. subtilis WBG2, carrying an integrated groE-bgaB transcription fusion in the amyE locus, was mutagenized. Dark blue colonies formed at 37 degrees C represent mutants which constitutively produce BgaB (a thermostable beta-galactosidase) at high levels. Seven mutants (WBG101 to WBG107) were selected for further characterization. They all overproduced BgaB, GroEL, and DnaK simultaneously at 37 degrees C. These mutants could be restored to normal by introducing a plasmid carrying a functional copy of orf39, the first gene in the B. subtilis dnaK operon. Genomic sequencing of these mutants demonstrated that they all carried a single mutation in orf39. These mutations can be divided into three groups: (i) Gly-307 to Asp, (ii) Ser-122 to Phe, and (iii) Gly-63 to Glu. By using a binary vector system in E. coli, production of ORF39 was found to negatively regulate the expression of groE-bgaB in a CIRCE-specific manner. Under the heat shock condition, the negative regulation mediated by ORF39 was abolished. Mobility shift of the CIRCE-containing probe was also observed with the crude extract prepared from the E. coli strain that overproduced ORF39. Therefore, ORF39 is the negative regulatory factor which regulates both groE and dnaK expression in B. subtilis. It is likely to function as a CIRCE- specific repressor.

This article has been cited by other articles:

• Wolff, S., Otto, A., Albrecht, D., Zeng, J. S., Buttner, K., Gluckmann, M., Hecker, M., Becher, D. (2006). Gel-free and Gel-based Proteomics in Bacillus subtilis: A Comparative Study. Mol. Cell. Proteomics 5: 1183-1192 [Abstract] [Full Text]  
• Ventura, M., Zink, R., Fitzgerald, G. F., van Sinderen, D. (2005). Gene Structure and Transcriptional Organization of the dnaK Operon of Bifidobacterium breve UCC 2003 and Application of the Operon in Bifidobacterial Tracing. Appl. Environ. Microbiol. 71: 487-500 [Abstract] [Full Text]  
• Kock, H., Gerth, U., Hecker, M. (2004). The ClpP Peptidase Is the Major Determinant of Bulk Protein Turnover in Bacillus subtilis. J. Bacteriol. 186: 5856-5864 [Abstract] [Full Text]  
• Mader, U., Hennig, S., Hecker, M., Homuth, G. (2004). Transcriptional Organization and Posttranscriptional Regulation of the Bacillus subtilis Branched-Chain Amino Acid Biosynthesis Genes. J. Bacteriol. 186: 2240-2252 [Abstract] [Full Text]  
• Chastanet, A., Derre, I., Nair, S., Msadek, T. (2004). clpB, a Novel Member of the Listeria monocytogenes CtsR Regulon, Is Involved in Virulence but Not in General Stress Tolerance. J. Bacteriol. 186: 1165-1174 [Abstract] [Full Text]  
• Hitomi, M., Nishimura, H., Tsujimoto, Y., Matsui, H., 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 [Abstract] [Full Text]  
• Wilson, A. C., Tan, M. (2002). Functional Analysis of the Heat Shock Regulator HrcA of Chlamydia trachomatis. J. Bacteriol. 184: 6566-6571 [Abstract] [Full Text]  
• Darmon, E., Noone, D., Masson, A., Bron, S., Kuipers, O. P., Devine, K. M., Dijl, J. M. v. (2002). A Novel Class of Heat and Secretion Stress-Responsive Genes Is Controlled by the Autoregulated CssRS Two-Component System of Bacillus subtilis. J. Bacteriol. 184: 5661-5671 [Abstract] [Full Text]  
• Wu, S.-C., Yeung, J. C., Duan, Y., Ye, R., Szarka, S. J., Habibi, H. R., Wong, S.-L. (2002). Functional Production and Characterization of a Fibrin-Specific Single-Chain Antibody Fragment from Bacillus subtilis: Effects of Molecular Chaperones and a Wall-Bound Protease on Antibody Fragment Production. Appl. Environ. Microbiol. 68: 3261-3269 [Abstract] [Full Text]  
• 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 [Abstract] [Full Text]  
• Zuber, U., Drzewiecki, K., Hecker, M. (2001). Putative Sigma Factor SigI (YkoZ) of Bacillus subtilis Is Induced by Heat Shock. J. Bacteriol. 183: 1472-1475 [Abstract] [Full Text]  
• Melly, E., Setlow, P. (2001). Heat Shock Proteins Do Not Influence Wet Heat Resistance of Bacillus subtilis Spores. J. Bacteriol. 183: 779-784 [Abstract] [Full Text]  
• Watanabe, K., Yamamoto, T., Suzuki, Y. (2001). Renaturation of Bacillus thermoglucosidasius HrcA Repressor by DNA and Thermostability of the HrcA-DNA Complex In Vitro. J. Bacteriol. 183: 155-161 [Abstract] [Full Text]  
• Movahedi, S., Waites, W. (2000). A Two-Dimensional Protein Gel Electrophoresis Study of the Heat Stress Response of Bacillus subtilis Cells during Sporulation. J. Bacteriol. 182: 4758-4763 [Abstract] [Full Text]  
• Moch, C., Schrögel, O., Allmansberger, R. (2000). Transcription of the nfrA-ywcH Operon from Bacillus subtilis Is Specifically Induced in Response to Heat. J. Bacteriol. 182: 4384-4393 [Abstract] [Full Text]  
• Minder, A. C., Fischer, H.-M., Hennecke, H., Narberhaus, F. (2000). Role of HrcA and CIRCE in the Heat Shock Regulatory Network of Bradyrhizobium japonicum. J. Bacteriol. 182: 14-22 [Abstract] [Full Text]  
• Nakahigashi, K., Ron, E. Z., Yanagi, H., Yura, T. (1999). Differential and Independent Roles of a sigma 32 Homolog (RpoH) and an HrcA Repressor in the Heat Shock Response of Agrobacterium tumefaciens. J. Bacteriol. 181: 7509-7515 [Abstract] [Full Text]  
• Macario, A. J. L., Lange, M., Ahring, B. K., De Macario, E. C. (1999). Stress Genes and Proteins in the Archaea. Microbiol. Mol. Biol. Rev. 63: 923-967 [Abstract] [Full Text]  
• Walker, D. C., Girgis, H. S., Klaenhammer, T. R. (1999). The groESL Chaperone Operon of Lactobacillus johnsonii. Appl. Environ. Microbiol. 65: 3033-3041 [Abstract] [Full Text]  
• Thies, F. L., Karch, H., Hartung, H.-P., Giegerich, G. (1999). Cloning and Expression of the dnaK Gene of Campylobacter jejuni and Antigenicity of Heat Shock Protein 70. Infect. Immun. 67: 1194-1200 [Abstract] [Full Text]  
• Krüger, E., Hecker, M. (1998). The First Gene of the Bacillus subtilis clpC Operon, ctsR, Encodes a Negative Regulator of Its Own Operon and Other Class III Heat Shock Genes. J. Bacteriol. 180: 6681-6688 [Abstract] [Full Text]  
• Bonorino, C., Nardi, N. B., Zhang, X., Wysocki, L. J. (1998). Characteristics of the Strong Antibody Response to Mycobacterial Hsp70: A Primary, T Cell-Dependent IgG Response with no Evidence of Natural Priming or {gamma}{delta} T Cell Involvement. J. Immunol. 161: 5210-5216 [Abstract] [Full Text]  
• Grandvalet, C., Rapoport, G., Mazodier, P. (1998). hrcA, Encoding the Repressor of the groEL Genes in Streptomyces albus G, Is Associated with a Second dnaJ Gene. J. Bacteriol. 180: 5129-5134 [Abstract] [Full Text]  
• Koch, B., Kilstrup, M., Vogensen, F. K., Hammer, K. (1998). Induced Levels of Heat Shock Proteins in a dnaK Mutant of Lactococcus lactis. J. Bacteriol. 180: 3873-3881 [Abstract] [Full Text]  
• Wu, S.-C., Ye, R., Wu, X.-C., Ng, S.-C., Wong, S.-L. (1998). Enhanced Secretory Production of a Single-Chain Antibody Fragment from Bacillus subtilis by Coproduction of Molecular Chaperones. J. Bacteriol. 180: 2830-2835 [Abstract] [Full Text]  
• Mogk, A., Völker, A., Engelmann, S., Hecker, M., Schumann, W., Völker, U. (1998). Nonnative Proteins Induce Expression of the Bacillus subtilis CIRCE Regulon. J. Bacteriol. 180: 2895-2900 [Abstract] [Full Text]  
• Baldini, R. L., Avedissian, M., Gomes, S. L. (1998). The CIRCE Element and Its Putative Repressor Control Cell Cycle Expression of the Caulobacter crescentus groESL Operon. J. Bacteriol. 180: 1632-1641 [Abstract] [Full Text]  
• Scharf, C., Riethdorf, S., Ernst, H., Engelmann, S., Völker, U., Hecker, M. (1998). Thioredoxin Is an Essential Protein Induced by Multiple Stresses in Bacillus subtilis. J. Bacteriol. 180: 1869-1877 [Abstract] [Full Text]  
• Nakamoto, H., Tanaka, N., Ishikawa, N. (2001). A Novel Heat Shock Protein Plays an Important Role in Thermal Stress Management in Cyanobacteria. J. Biol. Chem. 276: 25088-25095 [Abstract] [Full Text]  
• Servant, P., Grandvalet, C., Mazodier, P. (2000). The RheA repressor is the thermosensor of the HSP18 heat shock response in Streptomyces albus. Proc. Natl. Acad. Sci. USA 97: 3538-3543 [Abstract] [Full Text]