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Journal of Bacteriology, July 2000, p. 3802-3808, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Fur Positive Regulation of Iron Superoxide Dismutase in Escherichia coli: Functional Analysis of the sodB Promoter

Sarah Dubrac and Danièle Touati*

Institut Jacques Monod, CNRS-Universités Paris 6 et Paris 7, 75251 Paris Cedex 05, France

Received 25 February 2000/Accepted 19 April 2000

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 beta -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.

* Corresponding author. Mailing address: Institut Jacques Monod, CNRS-Universités Paris 6 et Paris 7, 2 place Jussieu, 75251 Paris Cedex 05, France. Phone: 33 1 44 27 47 19. Fax: 33 1 44 27 76 67. E-mail: touatida{at}ccr.jussieu.fr.

Journal of Bacteriology, July 2000, p. 3802-3808, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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