Polarity in the glnA operon: suppression of the reg- phenotype by rho mutations.
To determine the ability of mutations in glnA, the gene for glutamine synthetase (GS), to regulate nitrogen assimilatory enzymes, we assayed histidase and GS in 34 glnA (Gln(-)) strains. Twenty-five glnA mutants were RegC, synthesizing high levels of histidase regardless of the availability of nitrogen, and nine were Reg(-), synthesizing low levels of histidase in medium containing either limiting or excess ammonia. rho mutations were introduced into strains containing glnA point mutations or insertions in glnA, glnL, glnG, or glnF. The Reg(-) phenotype of strains with glnA point mutations, but not those with glnA or glnF insertions, was altered by the presence of rho, suggesting that glnA (Reg(-)) mutations are polar and exert their phenotype by decreasing expression of glnL and glnG. Consistent with this view, no GS protein was detected by two-dimensional gel electrophoresis in glnA (Reg(-)) rho(+) or glnA (Reg(-)) rho double mutants, whereas GS protein was detected in cells of 10 of 11 glnA (RegC) strains. Since glnA (Reg(-)) rho double mutants synthesize constitutive levels of histidase, GS protein is not necessary for full expression of histidase. Mu d1 insertions in glnL, but not those in glnG, responded to the presence of a rho allele, presumably owing to elevated transcription into glnG from the Mu d1 prophage. Our results suggest that glnA (Reg(-)) alleles are polar mutations, and a rho-dependent termination site down-stream is postulated as the basis for the polar phenomenon. The data also indicate that, under some circumstances, a significant portion of glnL and glnG transcription is initiated at the glnA promoter.
D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005809:Genes, Regulator; D005969:Glutamate Dehydrogenase / Q000096:biosynthesis; D005970:Glutamate Synthase / Q000096:biosynthesis; D005974:Glutamate-Ammonia Ligase / Q000096:biosynthesis / Q000235:genetics*; D006638:Histidine Ammonia-Lyase / Q000096:biosynthesis; D009154:Mutation; D009876:Operon; D014176:Protein Biosynthesis; D012234:Rho Factor; D013489:Suppression, Genetic
D004274:DNA, Recombinant; D012234:Rho Factor; D005970:Glutamate Synthase; D005969:Glutamate Dehydrogenase; D006638:Histidine Ammonia-Lyase; D005974:Glutamate-Ammonia Ligase