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J Bacteriol, May 1998, p. 2579-2582, Vol. 180, No. 9
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Coordinated Repression In Vitro of the Divergent
fepA-fes Promoters of Escherichia coli by the
Iron Uptake Regulation (Fur) Protein
Lucía
Escolar,
José
Pérez-Martín, and
Víctor
de
Lorenzo*
Centro Nacional de Biotecnología,
CSIC, 28049 Madrid, Spain
Received 21 November 1997/Accepted 27 February 1998
The mechanism involved in transcriptional repression of the
fepA-fes divergent promoters of Escherichia
coli by the Fur (ferric uptake regulation) protein has been
examined in vitro. This DNA region includes a suboptimal and
single Fur-binding site with two divergent and overlapped
35/
10
hexamers. Comparison of transcription patterns generated with
runoff experiments in either the presence or the absence of heparin
showed that access of the RNA polymerase to the principal
35/
10
hexamers was fully prevented by Fur-Mn2+ bound to its
target site within the divergent promoter region. Neither RNA
polymerase bound to the fes and fepA promoters
could be displaced by Fur-Mn2+, nor could the bound
repressor be outcompeted by an excess of the enzyme. However, the
repressor blocked reinitiation as soon as the polymerase moved away
from the fes promoter during transcription. The spatial
distribution of regulatory elements within the DNA region allowed the
simultaneous binding of the RNA polymerase to the fes and
fepA promoters and their coordinate regulation regardless
of their different transcriptional activities. Comparisons with other
iron-regulated systems support a general mechanism for Fur-controlled
promoters that implies a direct competition between the polymerase and
the regulator for overlapping target sites in the DNA.
*
Corresponding author. Mailing address: Centro Nacional
de Biotecnología, CSIC, Campus de Cantoblanco, 28049 Madrid,
Spain. Phone: (341) 585-4536. Fax: (341) 585-4506. E-mail:
vdlorenzo{at}cnb.uam.es.
J Bacteriol, May 1998, p. 2579-2582, Vol. 180, No. 9
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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