Anaerobic inductions of active forms of superoxide dismutases in Escherichia coli.
Escherichia coli growing anaerobically respond to NO3- with a approximately 3-fold induction of active FeSOD and a approximately 5.5-fold induction of an inactive, but activatable form of MnSOD (pro-MnSOD). Paraquat, which mediates anaerobic electron flow to NO3-, increased the induction of pro-MnSOD to approximately 2.5-fold. Strains with defects in the SOD genes or which lacked nitrate reductase activity failed to accumulate active or pro-forms of SODs in response to NO3- +/- PQ++. Diamide caused anaerobic induction of active MnSOD and this effect was also observed in a glutathione-negative strain. These inductions required de novo synthesis of protein, even when cell content of pro-MnSOD had been elevated by exposure to NO3- +/- PQ++ prior to addition of diamide. These results indicate that oxidation of a cell component increases biosynthesis of the SOD gene product and this postulated oxidation can be caused by terminal electron acceptors, such as dioxygen or NO3-. In addition, it appears that insertion of the correct metal can be rate-limiting, leading to competition by other metals and to the accumulation of inactive, incorrectly substituted pro-forms. Metal insertion may be dependent upon the valence of the metal, which may be influenced, in turn, by the redox status of the cells. Diamide and redox active agents such as ferricyanide may thus allow anaerobic production of active MnSOD by favoring the production of a complexed form of Mn(III) which can compete favorably with other metal cations for the active site of nascent MnSOD.
Privalle
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Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Fridovich
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D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D003958:Diamide / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D007501:Iron; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008345:Manganese; D011994:Recombinant Proteins / Q000378:metabolism; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000235:genetics
D001426:Bacterial Proteins; D007527:Isoenzymes; D011994:Recombinant Proteins; D003958:Diamide; D008345:Manganese; D007501:Iron; D013482:Superoxide Dismutase