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Oxidative stress rapidly stabilizes promoter-proximal paused Pol II across the human genome.


ABSTRACT: Oxidative stress has pervasive effects on cells but how they respond transcriptionally upon the initial insult is incompletely understood. We developed a nuclear walk-on assay that semi-globally quantifies nascent transcripts in promoter-proximal paused RNA polymerase II (Pol II). Using this assay in conjunction with ChIP-Seq, in vitro transcription, and a chromatin retention assay, we show that within a minute, hydrogen peroxide causes accumulation of Pol II near promoters and enhancers that can best be explained by a rapid decrease in termination. Some of the accumulated polymerases slowly move or 'creep' downstream. This second effect is correlated with and probably results from loss of NELF association and function. Notably, both effects were independent of DNA damage and ADP-ribosylation. Our results demonstrate the unexpected speed at which a global transcriptional response can occur. The findings provide strong support for the residence time of paused Pol II elongation complexes being much shorter than estimated from previous studies.

SUBMITTER: Nilson KA 

PROVIDER: S-EPMC5737879 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature

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Oxidative stress rapidly stabilizes promoter-proximal paused Pol II across the human genome.

Nilson Kyle A KA   Lawson Christine K CK   Mullen Nicholas J NJ   Ball Christopher B CB   Spector Benjamin M BM   Meier Jeffery L JL   Price David H DH  

Nucleic acids research 20171101 19


Oxidative stress has pervasive effects on cells but how they respond transcriptionally upon the initial insult is incompletely understood. We developed a nuclear walk-on assay that semi-globally quantifies nascent transcripts in promoter-proximal paused RNA polymerase II (Pol II). Using this assay in conjunction with ChIP-Seq, in vitro transcription, and a chromatin retention assay, we show that within a minute, hydrogen peroxide causes accumulation of Pol II near promoters and enhancers that ca  ...[more]

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