This Article Full Text 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 Urata, M. Articles by Nojiri, H. Search for Related Content PubMed PubMed Citation Articles by Urata, M. Articles by Nojiri, H.

 Previous Article  |  Next Article 

Journal of Bacteriology, October 2004, p. 6815-6823, Vol. 186, No. 20
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.20.6815-6823.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Transcriptional Regulation of the ant Operon, Encoding Two-Component Anthranilate 1,2-Dioxygenase, on the Carbazole-Degradative Plasmid pCAR1 of Pseudomonas resinovorans Strain CA10

Masaaki Urata, Masatoshi Miyakoshi, Satoshi Kai, Kana Maeda, Hiroshi Habe, Toshio Omori, Hisakazu Yamane, and Hideaki Nojiri^*

Biotechnology Research Center, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, Japan

Received 29 June 2004/ Accepted 19 July 2004

The carbazole-degradative plasmid pCAR1 of Pseudomonas resinovorans strain CA10 has two gene clusters, carAaAaBaBbCAcAdDFE and antABC, which are involved in the conversions of carbazole to anthranilate and anthranilate to catechol, respectively. We proved that the antABC gene cluster, encoding two-component anthranilate 1,2-dioxygenase, constitutes a single transcriptional unit through Northern hybridization and reverse transcription-PCR (RT-PCR) analyses. The transcription start point of antA was mapped at 53 bp upstream point of its translation start point, and the –10 and –35 boxes were homologous to conserved ⁷⁰ recognition sequence. Hence the promoter of the ant operon was designated P_ant. 5' Deletion analyses using luciferase as a reporter showed that the region up to at least 70 bp from the transcription start point of antA was necessary for the activation of P_ant. Luciferase expression from P_ant was induced by anthranilate itself, but not by catechol. Two probable AraC/XylS-type regulatory genes found on pCAR1, open reading frame 22 (ORF22) and ORF23, are tandemly located 3.2 kb upstream of the antA gene. We revealed that the product of ORF23, designated AntR, is indispensable for the stimulation of P_ant in Pseudomonas putida cells. Northern hybridization and RT-PCR analyses revealed that another copy of P_ant, which is thought to be translocated about 2.1 kb upstream of the carAa gene as a consequence of the transposition of ISPre1, actually drives transcription of the carAa gene in the presence of anthranilate, indicating that both ant and car operons are simultaneously regulated by AntR.

Present address: Department of Industrial Chemistry, Shibaura Institute of Technology, Shibaura, Minato-ku, Tokyo 108-8548, Japan.

This article has been cited by other articles:

• Oglesby, A. G., Farrow, J. M. III, Lee, J.-H., Tomaras, A. P., Greenberg, E. P., Pesci, E. C., Vasil, M. L. (2008). The Influence of Iron on Pseudomonas aeruginosa Physiology: A REGULATORY LINK BETWEEN IRON AND QUORUM SENSING. J. Biol. Chem. 283: 15558-15567 [Abstract] [Full Text]  
• Miyakoshi, M., Shintani, M., Terabayashi, T., Kai, S., Yamane, H., Nojiri, H. (2007). Transcriptome Analysis of Pseudomonas putida KT2440 Harboring the Completely Sequenced IncP-7 Plasmid pCAR1. J. Bacteriol. 189: 6849-6860 [Abstract] [Full Text]  
• Shintani, M., Urata, M., Inoue, K., Eto, K., Habe, H., Omori, T., Yamane, H., Nojiri, H. (2007). The Sphingomonas Plasmid pCAR3 Is Involved in Complete Mineralization of Carbazole. J. Bacteriol. 189: 2007-2020 [Abstract] [Full Text]  
• Shintani, M., Yano, H., Habe, H., Omori, T., Yamane, H., Tsuda, M., Nojiri, H. (2006). Characterization of the Replication, Maintenance, and Transfer Features of the IncP-7 Plasmid pCAR1, Which Carries Genes Involved in Carbazole and Dioxin Degradation.. Appl. Environ. Microbiol. 72: 3206-3216 [Abstract] [Full Text]  
• Inoue, K., Habe, H., Yamane, H., Nojiri, H. (2006). Characterization of Novel Carbazole Catabolism Genes from Gram-Positive Carbazole Degrader Nocardioides aromaticivorans IC177.. Appl. Environ. Microbiol. 72: 3321-3329 [Abstract] [Full Text]  
• Kang, M.-S., Besser, T. E., Call, D. R. (2006). Variability in the Region Downstream of the blaCMY-2 {beta}-Lactamase Gene in Escherichia coli and Salmonella enterica Plasmids.. Antimicrob. Agents Chemother. 50: 1590-1593 [Abstract] [Full Text]  
• Miyakoshi, M., Urata, M., Habe, H., Omori, T., Yamane, H., Nojiri, H. (2006). Differentiation of Carbazole Catabolic Operons by Replacement of the Regulated Promoter via Transposition of an Insertion Sequence. J. Biol. Chem. 281: 8450-8457 [Abstract] [Full Text]  
• Carl, B., Fetzner, S. (2005). Transcriptional Activation of Quinoline Degradation Operons of Pseudomonas putida 86 by the AraC/XylS-Type Regulator OxoS and Cross-Regulation of the PqorM Promoter by XylS. Appl. Environ. Microbiol. 71: 8618-8626 [Abstract] [Full Text]