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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Activation of the Escherichia coli nitrate reductase (narGHJI) operon by NarL and Fnr requires integration host factor.

Integration host factor protein (IHF) was shown to be required for Fnr- and NarL-dependent activation of the nitrate reductase (narGHJI) operon of Escherichia coli in response to nitrate availability and anaerobiosis. Using a narG- lacZ reporter fusion to evaluate narGHJI expression in vivo both the nitrate and anaerobic dependent controls were severely impaired in a himA mutant compared with the wild type strain. IHF was also required for Fnr-independent anaerobic control of narGHJI expression. In vitro, purified IHF protein was shown to bind to a narG promoter fragment with an apparent dissociation value of 5 nM by use of a gel shift assay. DNase I footprinting studies revealed that IHF protects a 37-base pair region centered 125 base pairs 5' of the narG transcription site. These studies suggest that the IHF protein performs a DNA bending function at the narG promoter to allow nitrate-dependent activation by the NarL regulatory protein, and second, it enhances the Fnr-dependent expression from the narG promoter under anaerobic cell growth conditions. A model whereby three transcriptional activators, NarL, IHF, and Fnr, induce expression of a sigma 70-dependent promoter for the narGHJI operon is discussed.[1]


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