Transcriptional and posttranscriptional regulation of manganese superoxide dismutase biosynthesis in Escherichia coli, studied with operon and protein fusions.
Protein and operon fusions between the manganese superoxide dismutase (MnSOD) gene, sodA, and genes of the lactose operon were constructed in an attempt to explore the effects of various factors on MnSOD expression and the level at which they operate. In sodA-lacZ protein fusions, induction of beta-galactosidase perfectly mimicked MnSOD induction (i.e., beta-galactosidase was not expressed in anaerobiosis and was induced by oxygen, redox-cycling compounds in aerobiosis, and iron chelators in anaerobiosis). In tac-sodA operon fusions, MnSOD induction was monitored only by the lactose operon inducer isopropyl-beta-D-thiogalactopyranoside. Various plasmids carrying part or all of the sodA regulatory and structural region inhibited aerobic beta-galactosidase induction in sodA-lacZ fusions. This included plasmids carrying only the transcription start and upstream region and also plasmids which did not contain this region and in which MnSOD was under foreign transcriptional control. The role of metal ions was also investigated. Addition of Mn(II) enhanced MnSOD activity but did not affect induction. The anaerobic expression of MnSOD from the oxygen-insensitive tac promoter was enhanced threefold by iron-chelating agents, implying a posttranscriptional or most likely a posttranslational modulation of enzyme activity via metal ions. To accommodate all these data, multiregulation of MnSOD is proposed.
D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology*; D005838:Genotype; D008345:Manganese; D009154:Mutation; D009876:Operon; D010618:Phenanthrolines; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase / Q000096:biosynthesis*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D010618:Phenanthrolines; D011993:Recombinant Fusion Proteins; D008345:Manganese; D013482:Superoxide Dismutase; D001616:beta-Galactosidase; C025205:1,10-phenanthroline