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 McFall, S. M. Articles by Chakrabarty, A. M. Search for Related Content PubMed PubMed Citation Articles by McFall, S. M. Articles by Chakrabarty, A. M.

 Previous Article  |  Next Article 

J. Bacteriol., 06 1997, 3655-3663, Vol 179, No. 11
Copyright © 1997, American Society for Microbiology

2-chloromuconate and ClcR-mediated activation of the clcABD operon: in vitro transcriptional and DNase I footprint analyses

SM McFall, MR Parsek and AM Chakrabarty
Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago 60612, USA.

In Pseudomonas putida, the plasmid-borne clcABD operon encodes enzymes involved in 3-chlorocatechol degradation. Previous studies have demonstrated that these enzymes are induced when P. putida is grown in the presence of 3-chlorobenzoate, which is converted to 3- chlorocatechol, and that ClcR, a LysR-type regulator, is required for this induction. The clcABD operon is believed to have evolved from the chromosomal catBCA operon, which encodes enzymes that utilize catechol and is regulated by CatR. The inducer for the catBCA operon is an intermediate of the catechol pathway, cis,cis-muconate. In this study, we demonstrate by the use of in vitro transcription assays and lacZ transcription fusions in vivo that the analogous intermediate of the 3- chlorocatechol pathway, 2-chloromuconate, is the inducer of the clcABD operon. The DNase I footprints of ClcR with and without 2- chloromuconate were also determined. An extended region of the promoter from -79 to -25 was occupied in the absence of inducer, but the -35 region was unprotected. When 2-chloromuconate was added to the binding assays, the footprint contracted approximately 4 bp at the proximal end of the promoter, and the -35 region was contacted. It is interesting to note that CatR actually extends its footprint 14 bp on the catBCA promoter in response to its inducer. Although CatR and ClcR change their nucleotide protection patterns in different manners when exposed to their respective inducers, their final footprints resemble each other. Therefore, it is possible that their transcriptional activation mechanisms may be evolutionarily conserved.

This article has been cited by other articles:

• Dubbs, P., Dubbs, J. M., Tabita, F. R. (2004). Effector-Mediated Interaction of CbbRI and CbbRII Regulators with Target Sequences in Rhodobacter capsulatus. J. Bacteriol. 186: 8026-8035 [Abstract] [Full Text]  
• Tropel, D., van der Meer, J. R. (2004). Bacterial Transcriptional Regulators for Degradation Pathways of Aromatic Compounds. Microbiol. Mol. Biol. Rev. 68: 474-500 [Abstract] [Full Text]  
• Corbella, M. E., Puyet, A. (2003). Real-Time Reverse Transcription-PCR Analysis of Expression of Halobenzoate and Salicylate Catabolism-Associated Operons in Two Strains of Pseudomonas aeruginosa. Appl. Environ. Microbiol. 69: 2269-2275 [Abstract] [Full Text]  
• Perez-Pantoja, D., Ledger, T., Pieper, D. H., Gonzalez, B. (2003). Efficient Turnover of Chlorocatechols Is Essential for Growth of Ralstonia eutropha JMP134(pJP4) in 3-Chlorobenzoic Acid. J. Bacteriol. 185: 1534-1542 [Abstract] [Full Text]  
• Bundy, B. M., Collier, L. S., Hoover, T. R., Neidle, E. L. (2002). Synergistic transcriptional activation by one regulatory protein in response to two metabolites. Proc. Natl. Acad. Sci. USA 99: 7693-7698 [Abstract] [Full Text]  
• Rapp, P. (2001). Multiphasic Kinetics of Transformation of 1,2,4-Trichlorobenzene at Nano- and Micromolar Concentrations by Burkholderia sp. Strain PS14. Appl. Environ. Microbiol. 67: 3496-3500 [Abstract] [Full Text]  
• Klemba, M., Jakobs, B., Wittich, R.-M., Pieper, D. (2000). Chromosomal Integration of tcb Chlorocatechol Degradation Pathway Genes as a Means of Expanding the Growth Substrate Range of Bacteria To Include Haloaromatics. Appl. Environ. Microbiol. 66: 3255-3261 [Abstract] [Full Text]  
• Arai, H., Ohishi, T., Chang, M. Y., Kudo, T. (2000). Arrangement and regulation of the genes for meta-pathway enzymes required for degradation of phenol in Comamonas testosteroni TA441. Microbiology 146: 1707-1715 [Abstract] [Full Text]  
• Ogawa, N., McFall, S. M., Klem, T. J., Miyashita, K., Chakrabarty, A. M. (1999). Transcriptional Activation of the Chlorocatechol Degradative Genes of Ralstonia eutropha NH9. J. Bacteriol. 181: 6697-6705 [Abstract] [Full Text]  
• Zago, A., Chugani, S., Chakrabarty, A. M. (1999). Cloning and Characterization of Polyphosphate Kinase and Exopolyphosphatase Genes from Pseudomonas aeruginosa 8830. Appl. Environ. Microbiol. 65: 2065-2071 [Abstract] [Full Text]  
• Potrawfke, T., Timmis, K. N., Wittich, R.-M. (1998). Degradation of 1,2,3,4-Tetrachlorobenzene by Pseudomonas chlororaphis RW71. Appl. Environ. Microbiol. 64: 3798-3806 [Abstract] [Full Text]