Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling

RNA Polymerase II (Pol II) transcriptional recycling is a mechanism for which the required factors and contributions to overall gene expression levels are poorly understood. We describe an in vitro methodology facilitating unbiased identification of putative RNA Pol II transcriptional recycling fact...

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Autores principales: Chen, Zhong, Hankey, William, Zhao, Yue, Groth, Jeff, Huang, Furong, Wang, Hongyan, Campos, Alexandre Rosa, Huang, Jiaoti, Roeder, Robert G., Wang, Qianben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8566496/
https://www.ncbi.nlm.nih.gov/pubmed/34732721
http://dx.doi.org/10.1038/s41467-021-26604-1
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author Chen, Zhong
Hankey, William
Zhao, Yue
Groth, Jeff
Huang, Furong
Wang, Hongyan
Campos, Alexandre Rosa
Huang, Jiaoti
Roeder, Robert G.
Wang, Qianben
author_facet Chen, Zhong
Hankey, William
Zhao, Yue
Groth, Jeff
Huang, Furong
Wang, Hongyan
Campos, Alexandre Rosa
Huang, Jiaoti
Roeder, Robert G.
Wang, Qianben
author_sort Chen, Zhong
collection PubMed
description RNA Polymerase II (Pol II) transcriptional recycling is a mechanism for which the required factors and contributions to overall gene expression levels are poorly understood. We describe an in vitro methodology facilitating unbiased identification of putative RNA Pol II transcriptional recycling factors and quantitative measurement of transcriptional output from recycled transcriptional components. Proof-of-principle experiments identified PAF1 complex components among recycling factors and detected defective transcriptional output from Pol II recycling following PAF1 depletion. Dynamic ChIP-seq confirmed PAF1 silencing triggered defective Pol II recycling in human cells. Prostate tumors exhibited enhanced transcriptional recycling, which was attenuated by antibody-based PAF1 depletion. These findings identify Pol II recycling as a potential target in cancer and demonstrate the applicability of in vitro and cellular transcription assays to characterize Pol II recycling in other disease states.
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spelling pubmed-85664962021-11-19 Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling Chen, Zhong Hankey, William Zhao, Yue Groth, Jeff Huang, Furong Wang, Hongyan Campos, Alexandre Rosa Huang, Jiaoti Roeder, Robert G. Wang, Qianben Nat Commun Article RNA Polymerase II (Pol II) transcriptional recycling is a mechanism for which the required factors and contributions to overall gene expression levels are poorly understood. We describe an in vitro methodology facilitating unbiased identification of putative RNA Pol II transcriptional recycling factors and quantitative measurement of transcriptional output from recycled transcriptional components. Proof-of-principle experiments identified PAF1 complex components among recycling factors and detected defective transcriptional output from Pol II recycling following PAF1 depletion. Dynamic ChIP-seq confirmed PAF1 silencing triggered defective Pol II recycling in human cells. Prostate tumors exhibited enhanced transcriptional recycling, which was attenuated by antibody-based PAF1 depletion. These findings identify Pol II recycling as a potential target in cancer and demonstrate the applicability of in vitro and cellular transcription assays to characterize Pol II recycling in other disease states. Nature Publishing Group UK 2021-11-03 /pmc/articles/PMC8566496/ /pubmed/34732721 http://dx.doi.org/10.1038/s41467-021-26604-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Chen, Zhong
Hankey, William
Zhao, Yue
Groth, Jeff
Huang, Furong
Wang, Hongyan
Campos, Alexandre Rosa
Huang, Jiaoti
Roeder, Robert G.
Wang, Qianben
Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title_full Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title_fullStr Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title_full_unstemmed Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title_short Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling
title_sort transcription recycling assays identify paf1 as a driver for rna pol ii recycling
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8566496/
https://www.ncbi.nlm.nih.gov/pubmed/34732721
http://dx.doi.org/10.1038/s41467-021-26604-1
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