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Originally published In Press as doi:10.1074/jbc.M010501200 on December 19, 2000

J. Biol. Chem., Vol. 276, Issue 12, 9478-9485, March 23, 2001
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A 72-Base Pair AT-rich DNA Sequence Element Functions as a Bacterial Gene Silencer*

Chien-Chung Chen, Ming Fang, Arundhati Majumder, and Hai-Young WuDagger

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.

* This work was supported by a National Institutes of Health Grant GM-53617.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF106956 and AF106955.

Dagger 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.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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