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Journal of Bacteriology, February 1999, p. 893-898, Vol. 181, No. 3
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cra-Dependent Transcriptional Activation of the icd Gene of Escherichia coli

Jean-François Prost,1 Didier Nègre,1 Christelle Oudot,1 Katsuhiko Murakami,2 Akira Ishihama,2 Alain J. Cozzone,1,* and Jean-Claude Cortay1

Institut de Biologie et Chimie des Protéines, Centre National de la Recherche Scientifique, Lyon, France,1 and National Institute of Genetics, Mishima, Shizuoka 411, Japan2

Received 29 June 1998/Accepted 20 November 1998

The icd gene of Escherichia coli, encoding isocitrate dehydrogenase, was shown to be expressed from two different promoters: the previously identified icd P1 and a newly detected second promoter, icd P2, whose expression is positively regulated by the catabolite repressor-activator protein Cra, formerly called FruR. In each case, we determined the mRNA start site by primer extension analysis of in vivo transcripts and examined the interaction of the icd control region with either RNA polymerase or Cra. We observed that (i) the Cra factor binds to and activates transcription from a site centered at position -76.5 within the icd P2 promoter region and (ii) three particular mutations in the C-terminal end of the alpha  subunit of RNA polymerase (L262A, R265A, and N268A) considerably diminish transcription initiating from the icd P2 promoter, as shown by in vitro experiments performed in the presence of mutant RNA polymerases carrying Ala substitutions.

* Corresponding author. Mailing address: Institut de Biologie et Chimie des Protéines, Centre National de la Recherche Scientifique, 7 Passage du Vercors, 69367 Lyon Cedex 07, France. Phone: 33 (0) 4.72.72.26.75. Fax: 33 (0) 4.72.72.26.01. E-mail: aj.cozzone{at}ibcp.fr.

Journal of Bacteriology, February 1999, p. 893-898, Vol. 181, No. 3
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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