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Journal of Bacteriology, December 1998, p. 6784-6788, Vol. 180, No. 24
Department of Microbiology, University of
Colorado Health Sciences Center, Denver, Colorado
80262,1 and
Department of
Microbiology and Immunology, Texas Tech University Health Sciences
Center, Lubbock, Texas 740302
Received 18 March 1998/Accepted 1 October 1998
We previously identified a novel regulator of the exotoxin A gene
(toxA) in Pseudomonas aeruginosa, PtxR, that
belongs to the LysR family of prokaryotic regulatory proteins.
Preliminary data also suggest that PtxR affects the expression of
siderophores in P. aeruginosa. Because toxA
expression and siderophore production in this organism are coordinately
regulated by the ferric uptake regulator (Fur) and the Fur-regulated
alternative sigma factor PvdS, regulation of ptxR itself in
the context of these regulators was examined. RNase protection analyses
of ptxR transcription revealed that there are two
independent transcription initiation sites (T1 and T2). While
transcription from the promoter of T1 is constitutive throughout the
growth cycle of PAO1, transcription from the second promoter (P2) is
negatively affected by iron. Transcription from the P2 promoter is
constitutive in a fur mutant under microaerobic conditions
but still iron regulated during aerobic growth. High concentrations
(>100 nM) of the ferric uptake regulatory protein (Fur) failed to bind
to either of the promoter regions of ptxR in either gel
mobility shift assays or DNase I footprint experiments. These results
indicate that Fur indirectly regulates the iron-dependent expression of
ptxR. Iron-regulated transcription of ptxR from
the P2 promoter, but not constitutive expression from the P1 promoter,
was dependent on the Fur-regulated alternative sigma factor gene
pvdS, even under aerobic conditions. Consequently, there
are two levels of iron-regulated expression of ptxR. The
iron-regulated expression of ptxR under microaerobic conditions from the P2 promoter of ptxR is mediated
indirectly by Fur through the iron-regulated expression of
pvdS. In contrast, pvdS-mediated iron
regulation of ptxR under aerobic conditions is Fur independent.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Fur-Regulated Gene Encoding the Alternative Sigma Factor PvdS
Is Required for Iron-Dependent Expression of the LysR-Type Regulator
PtxR in Pseudomonas aeruginosa
*
Corresponding author. Mailing address: Department of
Microbiology, Box B175, University of Colorado Health Sciences Center, 4200 E. Ninth Ave., Denver, CO 80262. Phone: (303) 315-8627. Fax: (303)
315-6785. E-mail: Mike.Vasil{at}uchsc.edu.
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