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J. Biol. Chem., Vol. 280, Issue 1, 585-595, January 7, 2005

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Identification of AcnR, a TetR-type Repressor of the Aconitase Gene acn in Corynebacterium glutamicum^*

Andreas Krug, Volker F. Wendisch, and Michael Bott

From the Institut für Biotechnologie 1, Forschungszentrum Jülich, D-52425 Jülich, Germany

In Corynebacterium glutamicum, the activity of aconitase is 2.5-4-fold higher on propionate, citrate, or acetate than on glucose. Here we show that this variation is caused by transcriptional regulation. In search for putative regulators, a gene (acnR) encoding a TetR-type transcriptional regulator was found to be encoded immediately downstream of the aconitase gene (acn) in C. glutamicum. Deletion of the acnR gene led to a 5-fold increased acn-mRNA level and a 5-fold increased aconitase activity, suggesting that AcnR functions as repressor of acn expression. DNA microarray analyses indicated that acn is the primary target gene of AcnR in the C. glutamicum genome. Purified AcnR was shown to be a homodimer, which binds to the acn promoter in the region from -11 to -28 relative to the transcription start. It thus presumably acts by interfering with the binding of RNA polymerase. The acn-acnR organization is conserved in all corynebacteria and mycobacteria with known genome sequence and a putative AcnR consensus binding motif (CAGNACnnncGTACTG) was identified in the corresponding acn upstream regions. Mutations within this motif inhibited AcnR binding. Because the activities of citrate synthase and isocitrate dehydrogenase were previously reported not to be increased during growth on acetate, our data indicate that aconitase is a major control point of tricarboxylic acid cycle activity in C. glutamicum, and they identify AcnR as the first transcriptional regulator of a tricarboxylic acid cycle gene in the Corynebacterianeae.

Received for publication, July 21, 2004 , and in revised form, October 5, 2004.

This article is dedicated to Prof. Rudolf K. Thauer on the occasion of his 65th birthday.

^* This work was supported by the European Union within the framework of the VALPAN project (QLK 3-2000-00497). 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.

To whom correspondence should be addressed: Institut für Biotechnologie 1, Forschungszentrum Jülich, D-52425 Jülich, Germany. Tel.: 49-2461-61-5515; Fax: 49-2461-61-2710; E-mail: m.bott{at}fz-juelich.de.

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