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The fix Escherichia coli region contains four genes related to carnitine metabolism

Authors

  • Knut Eichler,

    Corresponding author
    1. Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires. CNRS URA 1486, Institut National des Ciences Appliquées, F-69621 Villeurbanne Cedes. France
    Current affiliation:
    1. c/o Hoffmann-LaRoche AG, Bau 64/40, Grenzacherstr. 124, CH-4002 Basel, Switerland
    • c/o Hoffmann-La Roche AG, Bau 64/40, Grenzacherstr. 124, CH-4002 Basel, Switzerland
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  • Anne Buchet,

    1. Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires. CNRS URA 1486, Institut National des Ciences Appliquées, F-69621 Villeurbanne Cedes. France
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  • Fabienne Bourgis,

    1. Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires. CNRS URA 1486, Institut National des Ciences Appliquées, F-69621 Villeurbanne Cedes. France
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  • Hans-Peter Kleber,

    1. Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires. CNRS URA 1486, Institut National des Ciences Appliquées, F-69621 Villeurbanne Cedes. France
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    • Bereich Biochemie. Fakultät für Biowissenschaften, Pharmazie und Psychologie, Universität Leipzig. D-04 103 Leipzig, FRG

  • Marie-Andrée Mandrand-Berthelot

    1. Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires. CNRS URA 1486, Institut National des Ciences Appliquées, F-69621 Villeurbanne Cedes. France
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Abstract

Anaerobic carnitine metabolism in Escherichia coli was recently shown to involve six genes organized in the cai operon and located at the first minute on the chromosome. The DNA sequence lying at the 5′ end of the cai locus was further investigated. It contains four open reading frames organized as an operon. In vivo overexpression of this DNA region revealed four polypeptides with apparent molecular masses of 27, 33, 45 and 6 kDa. These proteins displayed significant amino acid sequence homologies with polypeptides encoded by the fixABCX operons from Azorhizobium caulinodans and Rhizobium meliloti. The four ORFs were thus named fixABCX. The first two gene products were also found to share a high degree of sequence similarity with the subunits β and α, respectively, of mammalian electron transfer flavoproteins, suggesting a role for these proteins in a redox reaction. A singly polycistronic 5 kb mRNA transcript was detected in Northern blots under anaerobic conditions in the presence of DL-carnitine. Expression of a fixA-lacZ transcriptional fusion was induced by L(-)-carnitine and crotonobetaine but not by D(+)-carnitine, γ-butyrobetaine, glycinebetaine and choline as found previously for the carnitine pathway. Similarly, the fix operon was repressed by glucose and nitrate. Moreover, expression of the fix operon was induced by the global regulatory proteins CRP and FNR and repressed by the histone-like protein H-NS. All these regulatory proteins have been shown also to control expression of carnitine enzymes. Results from Northern blots and lacZ fusion studies indicate a common regulation of expression of fix and cai operons, which implies a physiological linkage between these two loci.

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