PMID:9922253

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Citation

Prost, JF, Nègre, D, Oudot, C, Murakami, K, Ishihama, A, Cozzone, AJ and Cortay, JC (1999) Cra-dependent transcriptional activation of the icd gene of Escherichia coli. J. Bacteriol. 181:893-8

Abstract

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.

Links

PubMed PMC93456

Keywords

Amino Acid Substitution; Bacterial Proteins/metabolism; Base Sequence; DNA-Directed RNA Polymerases/metabolism; Deoxyribonuclease I; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Isocitrate Dehydrogenase/biosynthesis; Isocitrate Dehydrogenase/genetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Promoter Regions, Genetic; RNA, Messenger/genetics; Recombinant Proteins/metabolism; Regulatory Sequences, Nucleic Acid; Repressor Proteins/metabolism; Restriction Mapping; Transcription, Genetic; Transcriptional Activation

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