PMID:14759508
| Citation |
Barnard, AM, Lloyd, GS, Green, J, Busby, SJ and Lee, DJ (2004) Location of the Escherichia coli RNA polymerase alpha subunit C-terminal domain at an FNR-dependent promoter: analysis using an artificial nuclease. FEBS Lett. 558:13-8 |
|---|---|
| Abstract |
The Escherichia coli FNR protein is a global transcription regulator that activates gene expression via interactions with the RNA polymerase alpha subunit C-terminal domain. Using preparations of E. coli RNA polymerase holoenzyme, specifically labelled with a DNA cleavage reagent, we have determined the location and orientation of the C-terminal domain of the RNA polymerase alpha subunit in transcriptionally competent complexes at a class II FNR-dependent promoter. We conclude that one alpha subunit C-terminal domain binds immediately upstream of FNR, and that its position and orientation is the same as at similar promoters dependent on CRP, another E. coli transcription activator that is related to FNR. In complementary experiments, we show that the second alpha subunit C-terminal domain of RNA polymerase can be repositioned by upstream-bound CRP, but not by upstream-bound FNR. |
| Links |
PubMed Online version:10.1016/S0014-5793(03)01518-7 |
| Keywords |
Base Sequence; Binding Sites; Cyclic AMP Receptor Protein/metabolism; DNA-Directed RNA Polymerases/chemistry; DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/metabolism; Dimerization; Edetic Acid/analogs & derivatives; Edetic Acid/metabolism; Escherichia coli/enzymology; Escherichia coli Proteins/metabolism; Iron-Sulfur Proteins/metabolism; Models, Chemical; Models, Molecular; Molecular Sequence Data; Promoter Regions, Genetic; Protein Structure, Tertiary; Trans-Activators; Transcription Factors/metabolism; Transcriptional Activation |
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