Cited by 123Genomics and Chromatin DynamicsRNA Research and SplicingRNA and protein synthesis mechanisms

Widespread Backtracking by RNA Pol II Is a Major Effector of Gene Activation, 5′ Pause Release, Termination, and Transcription Elongation Rate

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Sheridan, Ryan M., Fong, Nova, D’Alessandro, Angelo, Bentley, David L.·2019·Molecular Cell

Abstract

In addition to phosphodiester bond formation, RNA polymerase II has an RNA endonuclease activity, stimulated by TFIIS, which rescues complexes that have arrested and backtracked. How TFIIS affects transcription under normal conditions is poorly understood. We identified backtracking sites in human cells using a dominant-negative TFIIS (TFIIS DN) that inhibits RNA cleavage and stabilizes backtracked complexes. Backtracking is most frequent within 2 kb of start sites, consistent with slow elongation early in transcription, and in 3' flanking regions where termination is enhanced by TFIIS DN, suggesting that backtracked pol II is a favorable substrate for termination. Rescue from backtracking by RNA cleavage also promotes escape from 5' pause sites, prevents premature termination of long transcripts, and enhances activation of stress-inducible genes. TFIIS DN slowed elongation rates genome-wide by half, suggesting that rescue of backtracked pol II by TFIIS is a major stimulus of elongation under normal conditions.

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Authors
Sheridan, R. M., Fong, N., D’Alessandro, A., & Bentley, D. L.
Affiliations
University of Colorado Anschutz (US)
Corresponding author
Bentley, David L.
Venue
Molecular Cell
Publisher
Elsevier BV
Published
2019-01
Volume
73
Issue
1
Pages
107-118.e4
ISSN
1097-2765 (ISSN-L)
OpenAlex
W2902183734
Cited by
123
References
70
Citations by year
2026: 7 · 2025: 13 · 2024: 22 · 2023: 13 · 2022: 12 · 2021: 27 · 2020: 16 · 2019: 12
FWCI
4.89
Type
journal article
Language
en
Open access
Locations
  • Molecular Cell (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institutes of Health (R35GM118051)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Keywords
TFIIS, transcription termination, gene activation, stress-inducible genes, 3' flanking region, RNA cleavage
MeSH terms
Animals, Gene Expression Regulation, Humans, Kinetics, Mutation, RNA, RNA Polymerase II, Transcriptional Activation, 3' Flanking Region, Transcriptional Elongation Factors, Mice, HEK293 Cells, RNA Cleavage, Transcription Elongation, Genetic, Transcription Termination, Genetic

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