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Journal of Bacteriology, January 2004, p. 98-103, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.98-103.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

NpdR, a Repressor Involved in 2,4,6-Trinitrophenol Degradation in Rhodococcus opacus HL PM-1

Dang P. Nga,1 Josef Altenbuchner,2 and Gesche S. Heiss1*

Institute of Microbiology,1 Institute of Industrial Genetics, University of Stuttgart, 70550 Stuttgart, Germany2

Received 12 June 2003/ Accepted 2 October 2003

Rhodococcus opacus HL PM-1 utilizes 2,4,6-trinitrophenol (picric acid) as a sole nitrogen source. The initial attack on picric acid occurs through two hydrogenation reactions. Hydride transferase II (encoded by npdI) and hydride transferase I (encoded by npdC) are responsible for the hydride transfers. Database searches with the npd genes have indicated the presence of a putative transcriptional regulator, npdR. Here, the npdR gene was expressed in Escherichia coli, and the protein was purified and shown to form a complex with intergenic regions between open reading frames A and B and between npdH and npdI within the npd gene cluster. A change in DNA-NpdR complex formation occurred in the presence of 2,4-dinitrophenol, picric acid, 2-chloro-4,6-dinitrophenol, and 2-methyl-4,6-dinitrophenol. By constructing a promoter-probe vector, we demonstrated that both intergenic regions caused the expression of reporter gene xylE. Hence, both of these regions contain promoters. A deletion mutant of R. opacus HL PM-1 was constructed in which part of npdR was deleted. The expression of npdI and npdC was induced by 2,4-dinitrophenol in the wild-type strain, while in the mutant these genes were constitutively expressed. Hence, NpdR is a repressor involved in picric acid degradation.

* Corresponding author. Mailing address: Institute of Microbiology, University of Stuttgart, Allmandring 31, 70550 Stuttgart, Germany. Phone: 49 711 685 5491. Fax: 49 711 685 5725. E-mail: gesche.heiss{at}po.uni-stuttgart.de.

Journal of Bacteriology, January 2004, p. 98-103, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.98-103.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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