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Regulation of transcription and promoter mapping of the structural genes for nitrogenase (nifHDK) of Azospirillum brasilense Sp7

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Summary

Transcription of the structural genes for nitrogenase (nifHDK) in Azospirillum brasilense Sp7 was analysed using Northern blots of total RNA extracted from cultures grown under nitrogen-fixing conditions. Hybridization with an internal nifH probe revealed two transcripts, a major one (by concentration) of 1.1 kb corresponding to nifH and a minor one of 5.6 kb corresponding to nifHDK. Hybridization with nifD or nifK probes revealed the minor transcript of 5.6 kb. This confirms that the nifHDK genes are organized as a single transcription unit and suggests regulation at the level of termination of transcription. The complete nucleotide sequence of nifH was established and the DNA region upstream of the initiation codon was analysed for transcription and translation signals. The nifH open reading frame (ORF) is preceded by an NtrA-dependent promoter and two elements homologous to upstream activator sequences (UAS) required for NifA-mediated activation in other diazotrophs. Promoter mapping with S1 nuclease revealed two start sites located 10 by and 40 by downstream of the NtrA-dependent promoter.

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Authors and Affiliations

  1. Unité de Physiologic Cellulaire and URA CNRS 1300, Département des Biotechnologies, Institut Pasteur, 25 rue du Dr. Roux, 75724, Paris Cedex 15, France

    Miklos de Zamaroczy, Frédéric Delorme & Claudine Elmerich

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  1. Miklos de Zamaroczy
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  2. Frédéric Delorme
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  3. Claudine Elmerich
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Communicated by H. Hennecke

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de Zamaroczy, M., Delorme, F. & Elmerich, C. Regulation of transcription and promoter mapping of the structural genes for nitrogenase (nifHDK) of Azospirillum brasilense Sp7. Molec. Gen. Genet. 220, 33–42 (1989). https://doi.org/10.1007/BF00260852

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