ArticleProceedings of the National Academy of Sciences of the United States of America2020
Dynamics of RNA polymerase II and elongation factor Spt4/5 recruitment during activator-dependent transcription.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 46 citations in OpenAlex.
- Spt5's central KOW domains and the Pol II stalk collaborate to regulate chromatin and 3'-end processing in Saccharomyces cerevisiae.G3 (Bethesda, Md.) · 2026Article
- The Promoter-Proximal Pause: A Decision Point Governing RNA Polymerase II Fate.Annual review of biochemistry · 2026Review
- Kinetic control of mammalian transcription elongation.Nature structural & molecular biology · 2026Article
- A mechanism of synergistic Mediator recruitment in RNA polymerase II transcription activation revealed by single-molecule fluorescence.Molecular cell · 2025Article
- Real-time visualization of reconstituted transcription reveals RNAPII activation mechanisms at single promoters.Cell reports · 2025Article
- Multiple structures of RNA polymerase II isolated from human nuclei by ChIP-CryoEM analysis.Nature communications · 2025Article
- Regulation of RNA polymerase II transcription through re-initiation and bursting.Molecular cell · 2025Review
- Single-molecule analysis of transcription activation: dynamics of SAGA coactivator recruitment.Nature structural & molecular biology · 2025Article
- Real-time visualization of reconstituted transcription reveals RNA polymerase II activation mechanisms at single promoters.bioRxiv : the preprint server for biology · 2025Article
- DSIF factor Spt5 coordinates transcription, maturation and exoribonucleolysis of RNA polymerase II transcripts.Nature communications · 2025Article
- NusG-Spt5 Transcription Factors: Universal, Dynamic Modulators of Gene Expression.Journal of molecular biology · 2025Review
- Chromatin endogenous cleavage provides a global view of yeast RNA polymerase II transcription kinetics.eLife · 2024Article
- Chromatin endogenous cleavage provides a global view of yeast RNA polymerase II transcription kinetics.bioRxiv : the preprint server for biology · 2024Article
- Single-molecule reconstruction of eukaryotic factor-dependent transcription termination.Nature communications · 2024Article
- Time will tell: comparing timescales to gain insight into transcriptional bursting.Trends in genetics : TIG · 2024Review
- Transcriptional activators in the early Drosophila embryo perform different kinetic roles.Cell systems · 2023Article
- Impact ofInternational journal of molecular sciences · 2022Review
- The pausing zone and control of RNA polymerase II elongation by Spt5: Implications for the pause-release model.Molecular cell · 2022Article
- A set of Saccharomyces cerevisiae integration vectors for fluorescent dye labeling of proteins.G3 (Bethesda, Md.) · 2022Article
- Eukaryotic gene regulation at equilibrium, or non?Current opinion in systems biology · 2022Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
In eukaryotes, RNA polymerase II (RNApII) transcribes messenger RNA from template DNA. Decades of experiments have identified the proteins needed for transcription activation, initiation complex assembly, and productive elongation. However, the dynamics of recruitment of these proteins to transcription complexes, and of the transitions between these steps, are poorly understood. We used multiwavelength single-molecule fluorescence microscopy to directly image and quantitate these dynamics in a budding yeast nuclear extract that reconstitutes activator-dependent transcription in vitro. A strong activator (Gal4-VP16) greatly stimulated reversible binding of individual RNApII molecules to template DNA. Binding of labeled elongation factor Spt4/5 to DNA typically followed RNApII binding, was NTP dependent, and was correlated with association of mRNA binding protein Hek2, demonstrating specificity of Spt4/5 binding to elongation complexes. Quantitative kinetic modeling shows that only a fraction of RNApII binding events are productive and implies a rate-limiting step, probably associated with recruitment of general transcription factors, needed to assemble a transcription-competent preinitiation complex at the promoter. Spt4/5 association with transcription complexes was slowly reversible, with DNA-bound RNApII molecules sometimes binding and releasing Spt4/5 multiple times. The average Spt4/5 residence time was of similar magnitude to the time required to transcribe an average length yeast gene. These dynamics suggest that a single Spt4/5 molecule remains associated during a typical transcription event, yet can dissociate from RNApII to allow disassembly of abnormally long-lived (i.e., stalled) elongation complexes.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.