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TitleCryo-EM structure of TFIIH/Rad4-Rad23-Rad33 in damaged DNA opening in nucleotide excision repair.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 3338, Year 2021
Publish dateJun 7, 2021
AuthorsTrevor van Eeuwen / Yoonjung Shim / Hee Jong Kim / Tingting Zhao / Shrabani Basu / Benjamin A Garcia / Craig D Kaplan / Jung-Hyun Min / Kenji Murakami /
PubMed AbstractThe versatile nucleotide excision repair (NER) pathway initiates as the XPC-RAD23B-CETN2 complex first recognizes DNA lesions from the genomic DNA and recruits the general transcription factor ...The versatile nucleotide excision repair (NER) pathway initiates as the XPC-RAD23B-CETN2 complex first recognizes DNA lesions from the genomic DNA and recruits the general transcription factor complex, TFIIH, for subsequent lesion verification. Here, we present a cryo-EM structure of an NER initiation complex containing Rad4-Rad23-Rad33 (yeast homologue of XPC-RAD23B-CETN2) and 7-subunit coreTFIIH assembled on a carcinogen-DNA adduct lesion at 3.9-9.2 Å resolution. A ~30-bp DNA duplex could be mapped as it straddles between Rad4 and the Ssl2 (XPB) subunit of TFIIH on the 3' and 5' side of the lesion, respectively. The simultaneous binding with Rad4 and TFIIH was permitted by an unwinding of DNA at the lesion. Translocation coupled with torque generation by Ssl2 and Rad4 would extend the DNA unwinding at the lesion and deliver the damaged strand to Rad3 (XPD) in an open form suitable for subsequent lesion scanning and verification.
External linksNat Commun / PubMed:34099686 / PubMed Central
MethodsEM (single particle)
Resolution3.9 - 9.25 Å
Structure data

EMDB-22576: TFIIH/Rad4-23-33 Complex in NER focused on Rad4-33
PDB-7m2u: Nucleotide Excision Repair complex TFIIH Rad4-33
Method: EM (single particle) / Resolution: 8.0 Å

EMDB-22587, PDB-7k01:
Structure of TFIIH in TFIIH/Rad4-Rad23-Rad33 DNA opening complex
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-22588, PDB-7k04:
Structure of TFIIH/Rad4-Rad23-Rad33/DNA in DNA opening
Method: EM (single particle) / Resolution: 9.25 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-SF4:
IRON/SULFUR CLUSTER

ChemComp-CA:
Unknown entry

Source
  • Saccharomyces cerevisiae (brewer's yeast)
  • Baker's yeast (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • synthetic construct (others)
KeywordsNUCLEAR PROTEIN/Transferase / TFIIH / Rad4 / Rad4/23 / XPC / NER / Nucleotide Excision Repair / GG-NER / NUCLEAR PROTEIN / NUCLEAR PROTEIN-Transferase complex / NUCLEAR PROTEIN/DNA / NUCLEAR PROTEIN-DNA complex

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