Open complex formation during transcription initiation at the Escherichia coli galP1 promoter:: the role of the RNA polymerase α subunit at promoters lacking an UP-element

We have studied the role of the C-terminal domain of the a subunit (alpha CTD) of Escherichia coli RNA polymerase during transcription initiation at promoters lacking an UP-element. The temperature requirement for open complex formation was used as an indication of the kinetics of this process. We have previously shown that alpha CTD is required for transcription initiation at low temperature at the galP1 promoter, a promoter containing an UP-element. DNase I footprinting has been used to reveal the structure of open promoter complexes and the temperature requirement for open complex formation has been determined using potassium permanganate as a probe. In this work we show that, although alpha CTD is not absolutely required for transcription initiation at promoters lacking an UP-element, it does play a role during transcription initiation. This role is independent of the sequence of the promoter upstream from the -35 region and does not require stable alpha CTD-DNA interactions as determined by DNase I footprinting. The role of alpha CTD at promoters lacking an UP-element is discussed.