Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair

Cell. 2020 Mar 19;180(6):1228-1244.e24. doi: 10.1016/j.cell.2020.02.010. Epub 2020 Mar 5.


Transcription-coupled nucleotide excision repair (TC-NER) is initiated by the stalling of elongating RNA polymerase II (RNAPIIo) at DNA lesions. The ubiquitination of RNAPIIo in response to DNA damage is an evolutionarily conserved event, but its function in mammals is unknown. Here, we identified a single DNA damage-induced ubiquitination site in RNAPII at RPB1-K1268, which regulates transcription recovery and DNA damage resistance. Mechanistically, RPB1-K1268 ubiquitination stimulates the association of the core-TFIIH complex with stalled RNAPIIo through a transfer mechanism that also involves UVSSA-K414 ubiquitination. We developed a strand-specific ChIP-seq method, which revealed RPB1-K1268 ubiquitination is important for repair and the resolution of transcriptional bottlenecks at DNA lesions. Finally, RPB1-K1268R knockin mice displayed a short life-span, premature aging, and neurodegeneration. Our results reveal RNAPII ubiquitination provides a two-tier protection mechanism by activating TC-NER and, in parallel, the processing of DNA damage-stalled RNAPIIo, which together prevent prolonged transcription arrest and protect against neurodegeneration.

Keywords: CRL/CSA/CSB; CS; ChIP-seq; Cockayne syndrome; NER; RNA polymerase II; RNAPII; TCR; TFIIH; UV-sensitive syndrome; UVSSA; UVsS; nucleotide excision repair; transcription coupled repair; ubiquitination of RNAPII.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • DNA / metabolism
  • DNA Damage / physiology
  • DNA Helicases / metabolism
  • DNA Repair / physiology*
  • DNA Repair Enzymes / genetics
  • DNA Repair Enzymes / metabolism
  • Female
  • HCT116 Cells
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism*
  • Ubiquitination


  • Carrier Proteins
  • UVSSA protein, human
  • DNA
  • RNA Polymerase II
  • Rpb1 protein, mouse
  • DNA Helicases
  • DNA Repair Enzymes