biostudies-literatureMevissen TETAmerican Cancer SocietyEMBOHoward Hughes Medical InstituteNIEHS NIH HHSNHLBI NIH HHSNational Institutes of Health7091-7106.e24https://www.ebi.ac.uk/biostudies/studies/S-EPMC11645862biostudies-literatureUnknown187(25)In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4^CSA, which cooperate with UVSSA and ELOF1 to recruit TFIIH. 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 requires STK19, a factor previously implicated in transcription recovery after UV exposure. A 1.9-Å cryo-electron microscopy structure shows that STK19 binds the TC-NER complex through CSA and the RPB1 subunit of RNA Pol II. Furthermore, AlphaFold predicts that STK19 interacts with the XPD subunit of TFIIH, and disrupting this interface impairs CellSTK19 positions TFIIH for cell-free transcription-coupled DNA repair.PMC11645862R01 HL098316DP2 ES036404Schmid EWWalter JCMevissen TETFarnung LKummecke MfalseSTK19 positions TFIIH for cell-free transcription-coupled DNA repair.In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4^CSA, which cooperate with UVSSA and ELOF1 to recruit TFIIH. 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 requires STK19, a factor previously implicated in transcription recovery after UV exposure. A 1.9-Å cryo-electron microscopy structure shows that STK19 binds the TC-NER complex through CSA and the RPB1 subunit of RNA Pol II. Furthermore, AlphaFold predicts that STK19 interacts with the XPD subunit of TFIIH, and disrupting this interface impairs 2024-01-01T00:00:00Z2024 Dec2026-06-02T10:45:48.794Z2025-04-04T01:54:56.636ZS-EPMC116458623954722810.1016/j.cell.2024.10.020