Klebsiella pneumoniae nifA product activates the Rhizobium meliloti nitrogenase promoter

Abstract

Bacteria in the genus Rhizobium normally fix nitrogen only when they interact with leguminous plants to produce on the roots a highly differentiated structure, the nodule, within which the bacteria differentiate into nitrogen-fixing bacteroids. By contrast, the enteric bacterium Klebsiella pneumoniae reduces nitrogen in a free-living state in conditions of low oxygen tension and deficiency of fixed nitrogen. In K. pneumoniae, the overall circuitry by which nitrogen-fixation (nif) genes are regulated has been elucidated. In response to ammonia starvation, the product of the glnG gene activates transcription of the nifLA operon; this activation is dependent on the product of glnF (ref. 4). The nifA gene product is in turn required for transcription of all the other nif genes, including the nifHDK operon which codes for the subunits of nitrogenase. In contrast, very little is known about the sequence of events involved in the regulated change in rhizobial nif gene expression associated with bacteroid differentiation. In the work described here, we identify the K. pneumoniae and Rhizobium meliloti nifHDK promoters by mapping the in vivo start points of transcription. By defining and comparing the DNA sequences of these two promoters, we find that they share an unexpected degree of homology. Further, by constructing fusions of each of the two promoters to the lacZ gene from Escherichia coli, we show that both promoters are activated by the product of the K. pneumoniae nifA gene.