title: Identification of a second gene involved in global regulation of fumarate reductase and other nitrate-controlled genes for anaerobic respiration in Escherichia coli.
abstract: Fumarate reductase catalyzes the final step of anaerobic electron transport in Escherichia coli when fumarate is used as a terminal electron acceptor. Transcription of the fumarate reductase operon (frdABCD) was repressed when cells were grown in the presence of either of the preferred terminal electron acceptors, oxygen or nitrate, and was stimulated modestly by fumarate. We have previously identified a locus called frdR which pleiotropically affects nitrate repression of fumarate reductase, trimethylamine N-oxide reductase, and alcohol dehydrogenase gene expression and nitrate induction of nitrate reductase expression (L. V. Kalman and R. P. Gunsalus, J. Bacteriol. 170:623-629, 1988). Transformation of various frdR mutants with plasmids identified two complementation groups, indicating that the frdR locus is composed of two genes. One class of mutants was not completely restored to wild-type frdA-lacZ expression or nitrate reductase induction when complemented with multicopy narX+ plasmids, whereas low-copy narX+ plasmid-containing strains were. A second class of frdR mutants was identified and shown to correspond to a previously described gene, narL (frdR2). Complementation of these strains with multicopy narL+ plasmids resulted in superrepression of frdA-lacZ expression and moderate elevation of nitrate reductase expression. Multicopy plasmids containing both narL+ and narX+ or only narL+ were able to complement narL mutants, whereas narX+ plasmids complemented narX mutants only when present in a copy number approximately equal to that of narL. Both narL and narX mutants retained normal oxygen control of frdA-lacZ expression. Both types of mutants are pleiotropic, as evidenced by derepressed levels of the fumarate reductase and trimethylamine N-oxide reductase enzymes and by defective induction of nitrate reductase when cells were grown in the presence of nitrate. These results indicate that both the narL and narX gene products must be present in a defined ratio in the cell. We conclude that these proteins interact to effect normal nitrate control of the anaerobic electron transport-associated operons. From these studies, we propose that narX encodes a nitrate sensor protein while narL encodes a DNA-binding regulatory protein which together function in a manner analogous to other two-component regulatory systems.
authors.affiliation: Department of Microbiology, University of California, Los Angeles.
authors.forename: L V
authors.initials: LV
authors.lastname: Kalman
authors.forename: R P
authors.initials: RP
authors.lastname: Gunsalus
mesh_terms: D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics*; D004579:Electron Transport; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D005786:Gene Expression Regulation; D009154:Mutation; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000096:biosynthesis / Q000235:genetics; D009566:Nitrates / Q000502:physiology*; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D013385:Succinate Dehydrogenase / Q000235:genetics* / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D009566:Nitrates; D013385:Succinate Dehydrogenase; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase; D009565:Nitrate Reductases; D050901:Nitrate Reductase