Structural basis for RNA polymerase II ubiquitylation and inactivation in transcription-coupled repair

  • Nat Struct Mol Biol. 2024 Mar;31(3):536-547. doi: 10.1038/s41594-023-01207-0.
Goran Kokic  1  2 ,  George Yakoub  3 ,  Diana van den Heuvel  3 ,  Annelotte P Wondergem  3 ,  Paula J van der Meer  3 ,  Yana van der Weegen  3 ,  Aleksandar Chernev  4 ,  Isaac Fianu  1 ,  Thornton J Fokkens  5 ,  Sonja Lorenz  5 ,  Henning Urlaub  4  6 ,  Patrick Cramer  7 ,  Martijn S Luijsterburg  8
Affiliations
  • 1. Department of Molecular Biology, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.
  • 2. Division of Structural Biology and Protein Therapeutics, Odyssey Therapeutics GmbH, Frankfurt am Main, Germany.
  • 3. Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
  • 4. Bioanalytical Mass Spectrometry, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.
  • 5. Ubiquitin Signaling Specificity, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.
  • 6. Bioanalytics Group, University Medical Center Göttingen, Institute of Clinical Chemistry, Göttingen, Germany.
  • 7. Department of Molecular Biology, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany. [email protected].
  • 8. Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. [email protected].
PMID: 38316879 DOI: 10.1038/s41594-023-01207-0
Abstract

During transcription-coupled DNA repair (TCR), RNA polymerase II (Pol II) transitions from a transcriptionally active state to an arrested state that allows for removal of DNA lesions. This transition requires site-specific ubiquitylation of Pol II by the CRL4CSA ubiquitin Ligase, a process that is facilitated by ELOF1 in an unknown way. Using cryogenic electron microscopy, biochemical assays and Cell Biology approaches, we found that ELOF1 serves as an adaptor to stably position UVSSA and CRL4CSA on arrested Pol II, leading to Ligase neddylation and activation of Pol II ubiquitylation. In the presence of ELOF1, a transcription factor IIS (TFIIS)-like element in UVSSA gets ordered and extends through the Pol II pore, thus preventing reactivation of Pol II by TFIIS. Our results provide the structural basis for Pol II ubiquitylation and inactivation in TCR.