WDR82/PNUTS-PP1 prevents transcription-replication conflicts by promoting RNA POLYMERASE II degradation on chromatin

Research output: Contribution to journalArticlepeer-review

Authors

  • Helga B. Landsverk
  • Lise E. Sandquist
  • Lilli T. E. Bay
  • Barbara Steurer
  • Coen Campsteijn
  • Ole J. B. Landsverk
  • Jurgen A. Marteijn
  • Laura Trinkle-Mulcahy
  • Randi G. Syljuasen

Colleges, School and Institutes

Abstract

Transcription-replication (T-R) conflicts cause replication stress and loss of genome integrity. However, the transcription-related processes that restrain such conflicts are poorly understood. Here, we demonstrate that the RNA polymerase II (RNAPII) C-terminal domain (CTD) phosphatase protein phosphatase 1 (PP1) nuclear targeting subunit (PNUTS)-PP1 inhibits replication stress. Depletion of PNUTS causes lower EdU uptake, S phase accumulation, and slower replication fork rates. In addition, the PNUTS binding partner WDR82 also promotes RNAPII-CTD dephosphorylation and suppresses replication stress. RNAPII has a longer residence time on chromatin after depletion of PNUTS or WDR82. Furthermore, the RNAPII residence time is greatly enhanced by proteasome inhibition in control cells but less so in PNUTS- or WDR82-depleted cells, indicating that PNUTS and WDR82 promote degradation of RNAPII on chromatin. Notably, reduced replication is dependent on transcription and the phospho-CTD binding protein CDC73 after depletion of PNUTS/WDR82. Altogether, our results suggest that RNAPII-CTD dephosphorylation is required for the continuous turnover of RNAPII on chromatin, thereby preventing T-R conflicts.

Details

Original languageEnglish
Article number108469
JournalCell Reports
Volume33
Issue number9
Publication statusPublished - 1 Dec 2020

Keywords

  • transcription-replication conflicts, replication stress, PP1 phosphatase, PNUTS, WDR82, RNA polymerase II, CTD phosphorylation, Cancer, CDC73, proteasome-mediated degradation