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STK19 positions TFIIH for cell-free transcription-coupled DNA repair.

Tycho E T MevissenMaximilian KümmeckeErnst W SchmidLucas FarnungJohannes C Walter
Published in: bioRxiv : the preprint server for biology (2024)
In transcription-coupled repair, stalled RNA polymerase II (Pol II) is recognized by CSB and CRL4 CSA , which co-operate with UVSSSA and ELOF1 to recruit TFIIH for nucleotide excision repair (TC-NER). To explore the mechanism of TC-NER, we recapitulated this reaction in vitro . When a plasmid containing a site-specific lesion is transcribed in frog egg extract, error-free repair is observed that depends on CSB, CRL4 CSA , UVSSA, and ELOF1. Repair also depends on STK19, a factor previously implicated in transcription recovery after UV exposure. A 1.9 Å cryo-electron microscopy structure shows that STK19 joins the TC-NER complex by binding CSA and the RPB1 subunit of Pol II. Furthermore, AlphaFold predicts that STK19 interacts with the XPD subunit of TFIIH, and disrupting this interface impairs cell-free repair. Molecular modeling suggests that STK19 positions TFIIH ahead of Pol II for lesion verification. In summary, our analysis of cell-free TC-NER suggests that STK19 couples RNA polymerase II stalling to downstream repair events.
Keyphrases
  • cell free
  • dna repair
  • circulating tumor
  • electron microscopy
  • transcription factor
  • dna damage
  • high resolution
  • binding protein