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J. Biol. Chem., Vol. 280, Issue 40, 34332-34342, October 7, 2005
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The Lactococcus lactis CodY Regulon
IDENTIFICATION OF A CONSERVED cis-REGULATORY ELEMENT*
1
From the
Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, Kerklaan 30, 9751 NN Haren and Friesland Foods Corporate Research, P. O. Box 87, 7400 AB Deventer, The Netherlands
CodY of Lactococcus lactis MG1363 is a transcriptional regulator that represses the expression of several genes encoding proteins of the proteolytic system. DNA microarray analysis, comparing the expression profiles of L. lactis MG1363 and an isogenic strain in which codY was mutated, was used to determine the CodY regulon. In peptide-rich medium and exponentially growing cells, where CodY exerts strong repressing activity, the expression of over 30 genes was significantly increased upon removal of codY. The differentially expressed genes included those predominantly involved in amino acid transport and metabolism. In addition, several genes belonging to other functional categories were derepressed, stressing the pleiotropic role of CodY. Scrutinizing the transcriptome data with bioinformatics tools revealed the presence of a novel over-represented motif in the upstream regions of several of the genes derepressed in L. lactis MG1363
codY. Evidence is presented that this 15-bp cis-sequence, AATTTTCWGAAAATT, serves as a high affinity binding site for CodY, as shown by electrophoretic mobility shift assays and DNase I footprinting analyses. The presence of this CodY-box is sufficient to evoke CodY-mediated regulation in vivo. A copy of this motif is also present in the upstream region of codY itself. It is shown that CodY regulates its own synthesis and requires the CodY-box and branched-chain amino acids to interact with its promoter.
Received for publication, March 2, 2005 , and in revised form, June 23, 2005.
* This work was supported by Friesland Foods Corporate Research and the Dutch Ministry of Economic Affairs (SENTER). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1 and Tables 1-5.
1 To whom correspondence should be addressed. Tel.: 31-50-363-2093; Fax: 31-50-363-2348; E-mail: O.P.Kuipers{at}rug.nl.
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