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

Author

Listed:
  • Zhong Chen

    (Duke University School of Medicine)

  • William Hankey

    (Duke University School of Medicine)

  • Yue Zhao

    (Duke University School of Medicine
    China Medical University)

  • 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)

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.

Suggested Citation

  • Zhong Chen & William Hankey & Yue Zhao & Jeff Groth & Furong Huang & Hongyan Wang & Alexandre Rosa Campos & Jiaoti Huang & Robert G. Roeder & Qianben Wang, 2021. "Transcription recycling assays identify PAF1 as a driver for RNA Pol II recycling," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-26604-1
    DOI: 10.1038/s41467-021-26604-1
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    References listed on IDEAS

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    1. Natalya Yudkovsky & Jeffrey A. Ranish & Steven Hahn, 2000. "A transcription reinitiation intermediate that is stabilized by activator," Nature, Nature, vol. 408(6809), pages 225-229, November.
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