7AD8

Core TFIIH-XPA-DNA complex with modelled p62 subunit

Summary for 7AD8

• Entry DOI: 10.2210/pdb7ad8/pdb
• Related: 6RO4
• EMDB information: 4970
• Descriptor: DNA (49-MER), General transcription factor IIH subunit 1, IRON/SULFUR CLUSTER, ... (12 entities in total)
• Functional Keywords: dna repair, transcription factor, dna binding protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 10
• Total formula weight: 440024.33

Authors

Koelmel, W.,Kuper, J.,Kisker, C. (deposition date: 2020-09-14, release date: 2021-07-28, Last modification date: 2024-05-15)

Primary citation

Barnett, J.T.,Kuper, J.,Koelmel, W.,Kisker, C.,Kad, N.M.
The TFIIH subunits p44/p62 act as a damage sensor during nucleotide excision repair.
Nucleic Acids Res., 48:12689-12696, 2020

PubMed Abstract: Nucleotide excision repair (NER) in eukaryotes is orchestrated by the core form of the general transcription factor TFIIH, containing the helicases XPB, XPD and five 'structural' subunits, p62, p44, p34, p52 and p8. Recent cryo-EM structures show that p62 makes extensive contacts with p44 and in part occupies XPD's DNA binding site. While p44 is known to regulate the helicase activity of XPD during NER, p62 is thought to be purely structural. Here, using helicase and adenosine triphosphatase assays we show that a complex containing p44 and p62 enhances XPD's affinity for dsDNA 3-fold over p44 alone. Remarkably, the relative affinity is further increased to 60-fold by dsDNA damage. Direct binding studies show this preference derives from p44/p62's high affinity (20 nM) for damaged ssDNA. Single molecule imaging of p44/p62 complexes without XPD reveals they bind to and randomly diffuse on DNA, however, in the presence of UV-induced DNA lesions these complexes stall. Combined with the analysis of a recent cryo-EM structure, we suggest that p44/p62 acts as a novel DNA-binding entity that enhances damage recognition in TFIIH. This revises our understanding of TFIIH and prompts investigation into the core subunits for an active role during DNA repair and/or transcription.

PubMed: 33166411
DOI: 10.1093/nar/gkaa973

Experimental method

ELECTRON MICROSCOPY (3.5 Å)