Anaerobic biosynthesis of the manganese-containing superoxide dismutase in Escherichia coli. Effects of diazenedicarboxylic acid bis(N,N'-dimethylamide) (diamide).
Anaerobically grown Escherichia coli accumulate active manganese-containing superoxide dismutase (MnSOD) upon exposure to diamide. This induction requires de novo biosynthesis of MnSOD. Catalase, glutathione disulfide reductase, and glucose-6-phosphate dehydrogenase were also induced by diamide in anaerobic E. coli. A GSH-negative strain of E. coli did not produce MnSOD under anaerobic conditions and was as responsive to diamide as was the wild type strain. Diamide which had been prereduced, by incubation with GSH, was ineffective. NO3- plus paraquat, which elicits increased anaerobic biosynthesis of the MnSOD polypeptide, but not of active MnSOD, synergized with diamide in the induction of active MnSOD. A similar increase in the ability of diamide to cause anaerobic biosynthesis of active MnSOD was seen when the production of the MnSOD polypeptide was increased by isopropyl-beta-D-thiogalactopyranoside, in a strain bearing the MnSOD gene under the control of the tac promoter. These results are explained in terms of a dual action of diamide, i.e. at both the transcriptional and the maturational levels of biosynthesis of MnSOD. Oxidative inactivation of an Fe(II)-containing repressor and oxidative facilitation of insertion of manganese, in place of iron, into the nascent MnSOD polypeptide, are the postulated bases of this dual action.
Privalle
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Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Fridovich
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D000693:Anaerobiosis; D015153:Blotting, Western; D003001:Cloning, Molecular; D003958:Diamide / Q000494:pharmacology*; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology*; D005978:Glutathione / Q000378:metabolism; D008970:Molecular Weight; D010957:Plasmids; D013482:Superoxide Dismutase / Q000096:biosynthesis*
D003958:Diamide; D013482:Superoxide Dismutase; D005978:Glutathione