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Nucleic Acids Research, 1987, Vol. 15, No. 20 8479-8499
© 1987

Articles

The symbiotic nitrogen fixation regulatory operon (fixRnifA) of Bradyrhizobium japonicum is expressed aerobically and is subject to a novel, nifA-independent type of activation

Beat Thony, Hans-Martin Fischer, Denise Anthamatten, Thomas Bruderer and Hauke Hennecke*

Mikrobiologisches Institut, Eidgenössische Technische Hochschule, Universitatstrasse 2 CH-8092 Zürich, Switzerland

*To whom reprint requests should be sent

Received August 14, 1987. Revised September 25, 1987. Accepted September 25, 1987.

The Bradyrhizobium japonicum N2 fixation regulatory gene, nifA, was sequenced and its transcription start site determined. Between the start of transcription and the nifA gene an open reading frame of 278 codons was found and named fixR. A deletion in fixR which allowed transcription into nifA resulted in a 50% reduced Fix activity. The fixRnifA operon was expressed 1n soybean root nodules, in cultures grown anaerobically with nitrate as terminal electron acceptor, in microaerobic cultures, and in aerobic cultures. The transcription start site (+1) was preceded by a characteristic nif(–24/–12)-type promoter consensus sequence. Double base-pair exchanges in the –12 but not in the –24 region resulted in a ‘promoter-down’ phenotype. A promoter-upstream DNA region between –50 and –148 was essential for maximal promoter activity. Expression from the promoter was not dependent on nifA. We conclude that the fixRnifA promoter is positively controlled, and that it requires a newly postulated transcriptional factor in order to become activated.


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