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Dynamic turnover of paused Pol II complexes at human promoters

Paused RNA polymerase II (Pol II) that piles up near most human promoters is the target of mechanisms that control entry into productive elongation. Whether paused Pol II is a stable or dynamic target remains unresolved. We report that most 5′ paused Pol II throughout the genome is turned over withi...

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Detalles Bibliográficos
Autores principales: Erickson, Benjamin, Sheridan, Ryan M., Cortazar, Michael, Bentley, David L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6120720/
https://www.ncbi.nlm.nih.gov/pubmed/30150253
http://dx.doi.org/10.1101/gad.316810.118
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author Erickson, Benjamin
Sheridan, Ryan M.
Cortazar, Michael
Bentley, David L.
author_facet Erickson, Benjamin
Sheridan, Ryan M.
Cortazar, Michael
Bentley, David L.
author_sort Erickson, Benjamin
collection PubMed
description Paused RNA polymerase II (Pol II) that piles up near most human promoters is the target of mechanisms that control entry into productive elongation. Whether paused Pol II is a stable or dynamic target remains unresolved. We report that most 5′ paused Pol II throughout the genome is turned over within 2 min. This process is revealed under hypertonic conditions that prevent Pol II recruitment to promoters. This turnover requires cell viability but is not prevented by inhibiting transcription elongation, suggesting that it is mediated at the level of termination. When initiation was prevented by triptolide during recovery from high salt, a novel preinitiated state of Pol II lacking the pausing factor Spt5 accumulated at transcription start sites. We propose that Pol II occupancy near 5′ ends is governed by a cycle of ongoing assembly of preinitiated complexes that transition to pause sites followed by eviction from the DNA template. This model suggests that mechanisms regulating the transition to productive elongation at pause sites operate on a dynamic population of Pol II that is turning over at rates far higher than previously suspected. We suggest that a plausible alternative to elongation control via escape from a stable pause is by escape from premature termination.
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spelling pubmed-61207202019-03-01 Dynamic turnover of paused Pol II complexes at human promoters Erickson, Benjamin Sheridan, Ryan M. Cortazar, Michael Bentley, David L. Genes Dev Research Paper Paused RNA polymerase II (Pol II) that piles up near most human promoters is the target of mechanisms that control entry into productive elongation. Whether paused Pol II is a stable or dynamic target remains unresolved. We report that most 5′ paused Pol II throughout the genome is turned over within 2 min. This process is revealed under hypertonic conditions that prevent Pol II recruitment to promoters. This turnover requires cell viability but is not prevented by inhibiting transcription elongation, suggesting that it is mediated at the level of termination. When initiation was prevented by triptolide during recovery from high salt, a novel preinitiated state of Pol II lacking the pausing factor Spt5 accumulated at transcription start sites. We propose that Pol II occupancy near 5′ ends is governed by a cycle of ongoing assembly of preinitiated complexes that transition to pause sites followed by eviction from the DNA template. This model suggests that mechanisms regulating the transition to productive elongation at pause sites operate on a dynamic population of Pol II that is turning over at rates far higher than previously suspected. We suggest that a plausible alternative to elongation control via escape from a stable pause is by escape from premature termination. Cold Spring Harbor Laboratory Press 2018-09-01 /pmc/articles/PMC6120720/ /pubmed/30150253 http://dx.doi.org/10.1101/gad.316810.118 Text en © 2018 Erickson et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Research Paper
Erickson, Benjamin
Sheridan, Ryan M.
Cortazar, Michael
Bentley, David L.
Dynamic turnover of paused Pol II complexes at human promoters
title Dynamic turnover of paused Pol II complexes at human promoters
title_full Dynamic turnover of paused Pol II complexes at human promoters
title_fullStr Dynamic turnover of paused Pol II complexes at human promoters
title_full_unstemmed Dynamic turnover of paused Pol II complexes at human promoters
title_short Dynamic turnover of paused Pol II complexes at human promoters
title_sort dynamic turnover of paused pol ii complexes at human promoters
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6120720/
https://www.ncbi.nlm.nih.gov/pubmed/30150253
http://dx.doi.org/10.1101/gad.316810.118
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