Fur positive regulation of iron superoxide dismutase in Escherichia coli:: Functional analysis of the sodB promoter

In Escherichia coli, the expression of sodB, which encodes iron superoxide dismutase, has been suggested to be activated by Fur, the iron-responsive global regulator initially characterized as a transcriptional repressor. We investigated sodB regulation by functional analysis of the sodB promoter using sodB-lac fusions with various truncated and mutated promoters. Several cis- and trans acting elements involved in sodB regulation have been identified, The P-galactosidase activity of sodB-lacZ reporter fusions and RNA analysis showed sevenfold iron dependent, Fur-mediated activation of expression. A region just downstream from -10, including a large palindromic sequence encompassing the +1 position followed by a 14-bp AT rich motif, is the site of Fur positive regulation, and the integrity of both sequences was required for full Fur-mediated activation. The life span of sodB mRNA was three times longer in a fur(+) strain, indicating that Fur-mediated activation proceeds, at least in part, at the posttranscriptional level. The H-NS and IHF histone-like factors also affected sodB expression. IHF slightly repressed sodB expression independently of Fur regulation. In contrast, H-NS negative regulation operated only in the absence of Fur. Remarkably, psodB behaved like a "pure extended -10" promoter. Deletion of the -35 region did not affect expression, whereas expression was totally abolished by a TG-to-CC mutation in the extended -10 sequence TGcTACCCT.