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<p>The general transcription machinery and its occupancy at promoters are highly conserved across metazoans. This contrasts with the kinetics of mRNA production that considerably differ between model species such as <i>Drosophila</i> and mouse. The molecular basis for these kinetic differences is currently unknown. Here, we used Single-Molecule Footprinting to measure RNA Polymerase II (Pol II) occupancy, the fraction of DNA molecules bound, at promoters in mouse and <i>Drosophila</i> cell lines. Single-molecule data reveals that Pol II occupancy is on average 3–5 times more frequent at transcriptionally active <i>Drosophila</i> promoters than active mouse promoters. Kinetic modelling of the occupancy states suggests that these differences in Pol II occupancy are determined by the ratio between the transcription initiation and Pol II turnover rates. We used chemical perturbation of transcription initiation to determine Pol II turnover rate in both species. Integration of these data into the model shows that infrequent Pol II occupancy in mouse is explained by the combination of high Pol II turnover and low transcription initiation rates.</p>

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Mouse promoters are characterised by low occupancy and high turnover of RNA polymerase II

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年3月31日

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  • Kasit Chatsirisupachai,
  • Christina J I Moene,
  • Rozemarijn Kleinendorst,
  • Elisa Kreibich,
  • Nacho Molina,
  • Arnaud Krebs

摘要

The general transcription machinery and its occupancy at promoters are highly conserved across metazoans. This contrasts with the kinetics of mRNA production that considerably differ between model species such as Drosophila and mouse. The molecular basis for these kinetic differences is currently unknown. Here, we used Single-Molecule Footprinting to measure RNA Polymerase II (Pol II) occupancy, the fraction of DNA molecules bound, at promoters in mouse and Drosophila cell lines. Single-molecule data reveals that Pol II occupancy is on average 3–5 times more frequent at transcriptionally active Drosophila promoters than active mouse promoters. Kinetic modelling of the occupancy states suggests that these differences in Pol II occupancy are determined by the ratio between the transcription initiation and Pol II turnover rates. We used chemical perturbation of transcription initiation to determine Pol II turnover rate in both species. Integration of these data into the model shows that infrequent Pol II occupancy in mouse is explained by the combination of high Pol II turnover and low transcription initiation rates.