1887

Abstract

Expression from the W operon promoter () is under a strict catabolic repression control mediated by the cAMP-catabolite repression protein (CRP) complex in a glucose-containing medium. The promoter is also activated by the integration host factor (IHF) and repressed by the specific transcriptional regulator HpaR when 4-hydroxyphenylacetate (4HPA) is not present in the medium. Expression from the promoter is also repressed in undefined rich medium such as LB, but the molecular basis of this mechanism is not understood. We present and studies to demonstrate the involvement of FIS protein in this catabolic repression. DNase I footprinting experiments show that FIS binds to multiple sites within the promoter. FIS-site I overlaps the CRP-binding site. By using an electromobility shift assay, we demonstrated that FIS efficiently competes with CRP for binding to the promoter, suggesting an antagonist/competitive mechanism. RT-PCR showed that the repression effect is relieved in a FIS deleted strain. The repression role of FIS at was further demonstrated by transcription assays. These results suggest that FIS contributes to silencing the promoter in the exponential phase of growth in an undefined rich medium when FIS is predominantly expressed.

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2008-07-01
2020-08-07
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