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The TFIIH subunits p44/p62 act as a damage sensor during nucleotide excision repair

Barnett, Jamie T., Kuper, Jochen, Koelmel, Wolfgang, Kisker, Caroline, Kad, Neil M (2020) The TFIIH subunits p44/p62 act as a damage sensor during nucleotide excision repair. Nucleic Acids Research, 48 (22). pp. 12689-12696. ISSN 0305-1048. (doi:10.1093/nar/gkaa973) (KAR id:86567)


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.

Item Type: Article
DOI/Identification number: 10.1093/nar/gkaa973
Subjects: Q Science > QD Chemistry > QD431 Organic Chemistry- Biochemistry- Proteins, peptides, amino acids
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Neil Kad
Date Deposited: 11 Feb 2021 16:37 UTC
Last Modified: 04 Mar 2024 15:17 UTC
Resource URI: (The current URI for this page, for reference purposes)

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