Previous Article | Next Article ![]()
Applied and Environmental Microbiology, September 1999, p. 4057-4063, Vol. 65, No. 9
Dipartimento di Genetica e di Biologia dei
Microrganismi, Università degli Studi di Milano, 20133 Milan,
Italy
Received 28 April 1999/Accepted 12 July 1999
Toluene-o-xylene monooxygenase is an enzymatic
complex, encoded by the touABCDEF genes, responsible for
the early stages of toluene and o-xylene degradation in
Pseudomonas stutzeri OX1. In order to identify the loci
involved in the transcriptional regulation of the tou gene
cluster, deletion analysis and complementation studies were carried out
with Pseudomonas putida PaW340 as a heterologous host
harboring pFB1112, a plasmid that allowed regulated expression, inducible by toluene and o-xylene and their corresponding
phenols, of the toluene-o-xylene monooxygenase. A locus
encoding a positive regulator, designated touR, was mapped
downstream from the tou gene cluster. TouR was found to be
similar to transcriptional activators of aromatic compound catabolic
pathways belonging to the NtrC family and, in particular, to DmpR (83%
similarity), which controls phenol catabolism. By using a
touA-C2,3O fusion reporter system and by primer extension
analysis, a TouR cognate promoter (PToMO) was
mapped, which showed the typical
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Identification of the Pseudomonas
stutzeri OX1 Toluene-o-Xylene Monooxygenase Regulatory
Gene (touR) and of Its Cognate Promoter
24 TGGC,
12 TTGC sequences
characteristic of
54-dependent promoters and putative
upstream activating sequences. By using the reporter system described,
we found that TouR responds to mono- and dimethylphenols, but not the
corresponding methylbenzenes. In this respect, the regulation of the
P. stutzeri system differs from that of other toluene or
xylene catabolic systems, in which the hydrocarbons themselves function
as effectors. Northern analyses indicated low transcription levels of
tou structural genes in the absence of inducers. Basal
toluene-o-xylene monooxygenase activity may thus transform
these compounds to phenols, which then trigger the TouR-mediated response.
*
Corresponding author. Mailing address: Dip. di Genetica
e di Biologia dei Microrganismi, Via Celoria, 26, 20133 Milan, Italy. Phone: (39) 02.266.05.227. Fax: (39) 02.266.45.51. E-mail:
Paola.Barbieri{at}unimi.it.
This article has been cited by other articles:
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»