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A soluble two-component regulatory system controls expression of quinoprotein ethanol dehydrogenase (QEDH) but not expression of cytochrome c₅₅₀ of the ethanol-oxidation system in Pseudomonas aeruginosa

Fachgebiet Technische Biochemie, Institut für Biotechnologie der Technischen Universität Berlin, Seestraße 13, D-13353 Berlin, Germany¹

Author for correspondence: Helmut Görisch. Tel: +49 30 31427582. Fax: +49 30 31427581. e-mail: Goerisch{at}lb.TU-Berlin.De

The regulation of the divergent promoters of the exaAB genes in Pseudomonas aeruginosa ATCC 17933, in which exaA encodes a quinoprotein ethanol dehydrogenase and exaB codes for a cytochrome c₅₅₀, was studied. Using transcriptional lacZ fusions, promoter activity during growth on several substrates was measured. These promoter-probe vectors were also used to identify regulatory mutants defective in exaAB induction. Transcription from both exaA and exaB was reduced significantly in four mutants. Two other mutants showed transcription from exaA that was reduced, but higher than wild-type transcription from exaB. The genes that are needed for exaA promoter induction were sequenced and found to encode a two-component regulatory system: a histidine sensor kinase, which lacks a transmembrane helical N-terminus and is presumably located in the cytoplasm, and a response regulator. The phenotypic characterization and restoration of the wild-type behaviour of the different regulatory mutants produced by different cosmids and subclones indicate that six different genes may be involved in regulating ethanol oxidation in P. aeruginosa.

Keywords: exaAB, regulation, sensor kinase, response regulator, divergent promoter

Abbreviations: Cb, carbenicillin; MM, minimal medium; PQQ, pyrroloquinoline quinone; QEDH, quinoprotein ethanol dehydrogenase; QMDH, quinoprotein methanol dehydrogenase; Tc, tetracycline

The GenBank accession number for the sequence reported in this work is AJ009858.2 (=CAB 95009.1).

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