Identification of the urease operon in Helicobacter pylori and its control by mRNA decay in response to pH

Junko K. Akada, Mutsunori Shirai, Hiroaki Takeuchi, Masataka Tsuda, Teruko Nakazawa

    Research output: Contribution to journalReview article

    101 Citations (Scopus)

    Abstract

    We investigated the transcription of the urease gene cluster ureABIEFGH in Helicobacter pylori to determine the regulation of gene expression of the highly produced enzyme urease. Northern blot hybridization analysis demonstrated that cells of the wild-type strain grown in an ordinary broth had transcripts of ureAB, ureABI, ureI, ureIE' and ure' FGH, but cells of a ureI-disrupted mutant had only the ureAB transcript. When the wild-type cells were exposed to pH 8 for 30 min, very little mRNA was detected. However, when exposed to pH 6, a large amount of the ureIE'' transcript, which was longer than the urelE' transcript, together with the additional transcripts ureABIEFGH and ure' EFGH were detected. Rifampicin addition experiments demonstrated that urease mRNAs, and the urelE' transcripts in particular, are more stable at pH 5.5 than at pH 7. In accord with these results, urease activity in the crude cell extract of the pH 5.5 culture was twice as much as that of the pH 7 culture, although the amounts of UreA and UreB detected by immunoblot analysis were similar. The transcription start point of ureI was identified by primer extension using a ureA promoter-deleted mutant, and a consensus sequence of RpoD-RNA polymerase was found in the ureI promoter. The 3' end of the ureIE' mRNA, determined using S1 nuclease mapping, revealed that the transcript is able to cover the majority of the ureE open reading frame (ORF) that might be sufficient for UreE activity. Based on the above results, we conclude that the urease gene cluster of H. pylori consists of two operons, ureAB and ureIEFGH, and that primary transcripts of the latter as well as the read-through transcript, ureABIEFGH, are cleaved to produce several species of mRNA. It has been suggested that the ureIEFGH operon is regulated post-transcriptionally by mRNA decay in response to environmental pH. We are tempted to speculate that the ureE'' transcript present in acidic pH may contribute to produce an active product that can proceed the nickel incorporation to the active centre, the final step of urease biosynthesis.

    Original languageEnglish
    Pages (from-to)1071-1084
    Number of pages14
    JournalMolecular Microbiology
    Volume36
    Issue number5
    DOIs
    Publication statusPublished - 2000 Jun 15

    ASJC Scopus subject areas

    • Microbiology
    • Molecular Biology

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