gltF, a member of the gltBDF operon of Escherichia coli, is involved in nitrogen-regulated gene expression.
We report here the construction and analysis of insertional mutations in each of the three genes of the gltBDF operon and the nucleotide sequence of the region downstream from gltD. Two open reading frames were identified, the first of which corresponds to gltF. The gltB and gltD genes code for the large and small subunits, respectively, of the enzyme glutamate synthase (GOGAT). gltF codes for a protein, with a molecular mass of 26,350 Da, which is required for Ntr induction. Histidase synthesis was determined as a measure of Ntr function. First, insertions in gltB, gltD or gltF all prevent Ntr induction. Second, complementation analysis indicates that high-level expression of both the gltD and gltF genes is required for the induction of the Ntr enzymes under nitrogen-limiting conditions, indicating that the phenotype of the gltB insertion probably results from polarity on gltD and gltF. Third, glutamate-dependent repression of the glt operon appears to be mediated by the product of the gltF gene. Thus, the gltBDF operon of Escherichia coli is involved in induction of the so-called Ntr enzymes in response to nitrogen deprivation, as well as in glutamate biosynthesis.
Castaño
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I
Departamento de Biología Molecular, Universidad Nacional Autónoma de México, Cuernavaca, Morelos.
Flores
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N
Valle
F
F
Covarrubias
A A
AA
Bolivar
F
F
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005970:Glutamate Synthase / Q000235:genetics; D006638:Histidine Ammonia-Lyase / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000494:pharmacology*; D016366:Open Reading Frames; D009876:Operon; D033903:Periplasmic Proteins; D010641:Phenotype
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D033903:Periplasmic Proteins; C078319:gltF protein, E coli; D005970:Glutamate Synthase; D006638:Histidine Ammonia-Lyase; D009584:Nitrogen