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TitleSTK19 positions TFIIH for cell-free transcription-coupled DNA repair.
Journal, issue, pagesCell, Vol. 187, Issue 25, Page 7091-7106.e24, Year 2024
Publish dateDec 12, 2024
AuthorsTycho E T Mevissen / Maximilian Kümmecke / Ernst W Schmid / Lucas Farnung / Johannes C Walter /
PubMed AbstractIn transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4, which cooperate with UVSSA and ELOF1 to recruit TFIIH. To explore the ...In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4, 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, 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 cell-free repair. Molecular modeling suggests that STK19 positions TFIIH ahead of RNA Pol II for lesion verification. Our analysis of cell-free TC-NER suggests that STK19 couples RNA Pol II stalling to downstream repair events.
External linksCell / PubMed:39547228 / PubMed Central
MethodsEM (single particle)
Resolution1.9 - 2.7 Å
Structure data

45050

9bz0

EMDB-45050, PDB-9bz0:
Structure of an STK19-containing TC-NER complex
Method: EM (single particle) / Resolution: 1.9 Å

EMDB-47261: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map IV)
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-47262: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map III)
Method: EM (single particle) / Resolution: 2.3 Å

EMDB-47263: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map II)
Method: EM (single particle) / Resolution: 2.1 Å

EMDB-47266: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map V)
Method: EM (single particle) / Resolution: 2.1 Å

EMDB-47267: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map VI)
Method: EM (single particle) / Resolution: 2.1 Å

EMDB-47271: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map VII)
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-47272: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map VIII)
Method: EM (single particle) / Resolution: 2.3 Å

EMDB-47273: Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (Consensus refinement)
Method: EM (single particle) / Resolution: 1.9 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
  • sus scrofa (pig)
  • synthetic construct (others)
KeywordsTRANSCRIPTION/DNA/RNA / DNA repair / RNA polymerase II / Co-transcriptional process / TRANSCRIPTION / TRANSCRIPTION-DNA-RNA complex

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