Abstract
The σ70 subunit of RNA polymerase plays an essential role in transcription initiation. In addition, σ70 has a critical regulatory role during transcription elongation at the bacteriophage λ late promoter, λPR′. At this promoter, σ70 mediates a pause in early elongation through contact with a DNA sequence element in the initially transcribed region that resembles a promoter −10 element. Here we provide evidence that σ70 also mediates a pause in early elongation at the lac promoter (plac). Like that at λPR′, the pause at plac is facilitated by a sequence element in the initially transcribed region that resembles a promoter −10 element. Using biophysical analysis, we demonstrate that the pause-inducing sequence element at plac stabilizes the interaction between σ70 and the remainder of the transcription elongation complex. Bioinformatic analysis suggests that promoter-proximal σ70-dependent pauses may play a role in the regulation of many bacterial promoters.
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Acknowledgements
We thank J. Roberts, K. Brodolin, A. Kapanidis and S. Weiss for discussion, S. Borukhov for purified GreA and GreB, A. Huerta and J. Collado-Vides for providing promoter sequences, S. Dove for comments on the manuscript, and R. Hellmiss for assistance with figure preparation. We also thank A. Hatoum and J. Roberts for sharing unpublished results. This work was supported by US National Institutes of Health grant GM44025 to A.H. and GM41376 to R.H.E. and by a Howard Hughes Medical Institute investigatorship to R.H.E.
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Nickels, B., Mukhopadhyay, J., Garrity, S. et al. The σ70 subunit of RNA polymerase mediates a promoter-proximal pause at the lac promoter. Nat Struct Mol Biol 11, 544–550 (2004). https://doi.org/10.1038/nsmb757
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DOI: https://doi.org/10.1038/nsmb757
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