STL-seq reveals pause-release and termination kinetics for promoter-proximal paused RNA polymerase II transcripts
| Autor: | Nicolle A. Rosa-Mercado, Daniele Canzio, Joshua T. Zimmer, Matthew D. Simon, Joan A. Steitz |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Osmosis
Kinetics RNA polymerase II Biology Article chemistry.chemical_compound Transcriptional regulation RNA Messenger Promoter Regions Genetic Molecular Biology Genome Sequence Analysis RNA Gene Expression Profiling RNA Promoter Cell Biology DNA Methylation Hormones Cell biology Ecdysterone Gene Expression Regulation Genetic Techniques chemistry Mutation biology.protein RNA Polymerase II Function (biology) DNA Protein Binding Signal Transduction |
| Zdroj: | Mol Cell |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2021.08.019 |
| Popis: | Summary Despite the critical regulatory function of promoter-proximal pausing, the influence of pausing kinetics on transcriptional control remains an active area of investigation. Here, we present Start-TimeLapse-seq (STL-seq), a method that captures the genome-wide kinetics of short, capped RNA turnover and reveals principles of regulation at the pause site. By measuring the rates of release into elongation and premature termination through the inhibition of pause release, we determine that pause-release rates are highly variable, and most promoter-proximal paused RNA polymerase II molecules prematurely terminate (∼80%). The preferred regulatory mechanism upon a hormonal stimulus (20-hydroxyecdysone) is to influence pause-release rather than termination rates. Transcriptional shutdown occurs concurrently with the induction of promoter-proximal termination under hyperosmotic stress, but paused transcripts from TATA box-containing promoters remain stable, demonstrating an important role for cis-acting DNA elements in pausing. STL-seq dissects the kinetics of pause release and termination, providing an opportunity to identify mechanisms of transcriptional regulation. |
| Databáze: | OpenAIRE |
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