Different physiological roles of two independent pathways for nitrite reduction to ammonia by enteric bacteria

Abstract

Operon fusion strains and mutants of Escherichia coli K-12 lacking the NADH-dependent nitrite reductase have been used to determine the regulation and physiological roles of two independent pathways for nitrite reduction to ammonia. Both the formate-and NADH-dependent pathways (Nrf and Nir, respectively) were totally repressed during aerobic growth, partially active during anaerobic growth in the absence of nitrite and further induced anaerobically by nitrite. Both were dependent upon a functional Fnr protein (a transcription activator of genes for anaerobic respiration). During anaerobic growth in the presence of nitrate, the Nir pathway was fully induced but Nrf was strongly repressed. Mutants defective in the NarL protein, which induces transcription of nitrate reductase genes but represses fumarate reductase genes in the presence of nitrate, were derepressed for Nrf activity during growth with nitrate, but the Nir enzyme was less active. The synthesis of Nrf components was also sensitive to glucose repression and weak activation by NarL during growth in the absence of nitrate. These data indicate that the Nir pathway provides a mechanism for detoxifying nitrite formed in the cytoplasm as a product of nitrate reduction. In contrast, the electrogenic reduction of nitrite by the Nrf pathway provides a secondary source of energy during anaerobic growth and is consequently repressed by the NarL protein when the thermodynamically more favourable electron acceptor, nitrate, is available. Two short DNA sequences, 5′-TACCAT-3′ and 5′-CTCCTT-3′, were found in the promoters of operons known to be activated or repressed by the NarL protein. It is proposed that NarL activates nir B transcription by binding to one or both of these sequences located 5′ to the RNA polymerase binding site, but represses other operons, including nrf, by binding close to the transcription start.