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

Zhong Chen (), William Hankey, Yue Zhao, Jeff Groth, Furong Huang, Hongyan Wang, Alexandre Rosa Campos, Jiaoti Huang, Robert G. Roeder and Qianben Wang ()
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Zhong Chen: Duke University School of Medicine
William Hankey: Duke University School of Medicine
Yue Zhao: Duke University School of Medicine
Jeff Groth: Duke University School of Medicine
Furong Huang: Duke University School of Medicine
Hongyan Wang: Duke University School of Medicine
Alexandre Rosa Campos: Sanford Burnham Prebys Medical Discovery Institute
Jiaoti Huang: Duke University School of Medicine
Robert G. Roeder: The Rockefeller University
Qianben Wang: Duke University School of Medicine

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract 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.

Date: 2021
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DOI: 10.1038/s41467-021-26604-1

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