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J. Biol. Chem., Vol. 276, Issue 12, 9478-9485, March 23, 2001
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From the Department of Pharmacology, School of Medicine, Wayne
State University, Detroit, Michigan 48201
We have previously demonstrated that sequential
activation of the bacterial ilvIH-leuO-leuABCD gene
cluster involves a promoter-relay mechanism. In the current study, we
show that the final activation of the leuABCD operon is
through a transcriptional derepression mechanism. The
leuABCD operon is transcriptionally repressed by the
presence of a 318-base pair AT-rich upstream element. LeuO is required
for derepressing the repressed leuABCD operon. Deletion analysis of the repressive effect of the 318-bp element has led to the
identification of a 72-bp AT-rich (78% A+T) DNA sequence element, AT4,
which is capable of silencing a number of unrelated promoters in
addition to the leuABCD promoter. AT4-mediated gene silencing is orientation-independent and occurs within a distance of
300 base pairs. Furthermore, an increased gene-silencing effect was
observed with a tandemly repeated AT4 dimer. The possible mechanism of
AT4-mediated gene silencing in bacteria is discussed.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF106956 and AF106955.
To whom correspondence should be addressed: Dept. of Pharmacology,
Wayne State University, School of Medicine, 540 E. Canfield Ave.,
Detroit, MI 48201. Tel.: 313-577-1584; Fax: 313-577-6739; E-mail:
haiwu@med.wayne.edu.
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