J Bacteriol, February 1998, p. 855-861, Vol. 180, No. 4
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina 27858-4354
Received 25 July 1997/Accepted 6 December 1997
The alc gene cluster of Bordetella
pertussis includes three genes, alcA,
alcB, and alcC, which are involved in alcaligin
siderophore biosynthesis in response to iron starvation. The production
of AlcA, AlcB, and AlcC in Bordetella cells and the
transcriptional organization of alcA, alcB, and
alcC were investigated by using a set of three
alc'-'lacZ gene fusion constructs that were contiguous with
the known promoter upstream of alcA and extended to fusion junctions within each alc cistron. All three
alc'-'lacZ fusions exhibited iron-repressible reporter gene
expression which was abolished by deletion of the 105-bp
alcA promoter-operator region. In an immunoblot analysis
using a monoclonal antibody specific for
-galactosidase, the
AlcA-LacZ, AlcB-LacZ, and AlcC-LacZ hybrid proteins were detected in
Bordetella cells grown under iron-depleted conditions. A
B. pertussis mutant in which the 105-bp alcA
promoter-operator region was deleted by allelic exchange was unable to
produce detectable levels of siderophore. Hybridization analysis using
gene-specific probes showed that alc-specific transcript
levels in the mutant were negligible compared with those of the
wild-type parent. These results confirm that alcA,
alcB, and alcC are cotranscribed from an
iron-regulated control region immediately upstream of alcA. Transcript analysis using hybridization probes representing regions downstream of alcC demonstrated that alc
transcription extends approximately 3.6 kb further downstream from the
alcC coding region, suggesting the cotranscription of
additional, uncharacterized alcaligin system genes.
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