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In Escherichia coli, the expression of a σ factor is expected to indirectly down-regulate the expression of genes recognized by another σ factor, due to σ factor competition for a limited pool of RNA polymerase core enzymes. Evidence suggests that the sensitivity of genes to indirect down-regulation differs widely. We studied the variability in this sensitivity in promoters primarily recognized by RNAP holoenzymes carrying σ(70). From qPCR and live single-cell, single-RNA measurements of the transcription kinetics of several σ(70)-dependent promoters in various conditions and from the analysis of σ factors population-dependent models of transcription initiation, we find that, the smaller is the time-scale of the closed complex formation relative to the open complex formation, the weaker is a promoter's responsiveness to changes in σ(38) numbers. We conclude that, in E. coli, a promoter's responsiveness to indirect regulation by σ factor competition is determined by the sequence-dependent kinetics of the rate limiting steps of transcription initiation.

作者:Vinodh K, Kandavalli;Huy, Tran;Andre S, Ribeiro

来源:Biochimica et biophysica acta 2016 年 1859卷 10期

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作者:
Vinodh K, Kandavalli;Huy, Tran;Andre S, Ribeiro
来源:
Biochimica et biophysica acta 2016 年 1859卷 10期
标签:
Closed Complex Formation In vivo Transcription dynamics Open Complex Formation Single RNA detection σ Factors competition
In Escherichia coli, the expression of a σ factor is expected to indirectly down-regulate the expression of genes recognized by another σ factor, due to σ factor competition for a limited pool of RNA polymerase core enzymes. Evidence suggests that the sensitivity of genes to indirect down-regulation differs widely. We studied the variability in this sensitivity in promoters primarily recognized by RNAP holoenzymes carrying σ(70). From qPCR and live single-cell, single-RNA measurements of the transcription kinetics of several σ(70)-dependent promoters in various conditions and from the analysis of σ factors population-dependent models of transcription initiation, we find that, the smaller is the time-scale of the closed complex formation relative to the open complex formation, the weaker is a promoter's responsiveness to changes in σ(38) numbers. We conclude that, in E. coli, a promoter's responsiveness to indirect regulation by σ factor competition is determined by the sequence-dependent kinetics of the rate limiting steps of transcription initiation.