The interaction of XPG with TFIIH through p62 and XPD is required for the completion of nucleotide excision repair

Nucleotide excision repair (NER) is the key pathway for the removal of DNA damage induced by UV irradiation and chemotherapeutic reagents. Protein-protein interactions are crucial for the dynamic and coordinated assembly of the proteins involved at DNA lesion. Here we focus on the role of interactions between the multi-subunit helicase/translocase complex TFIIH and the 3’ endonuclease XPG. We show that XPG interacts with the p62 and XPD subunits of TFIIH through its long spacer region located in the middle of its split active site. We show the interactions between three acidic regions of XPG with the pleckstrin homology (PH) domain of p62 are of moderate importance for NER, while interactions with XPD are critical for activity. Combining mutations in the p62 and XPD interaction domains leads to additive defects in NER activity. Unexpectedly, we show that these interactions are not required for the recruitment of XPG, but rather for the formation of a catalytically competent NER complex and for triggering the incision 5’ to the lesion by ERCC1-XPF. Our studies provide fundamental insights into how interactions between TFIIH and XPG contribute to the NER pathway and more generally how modular protein-protein interactions control each step along the NER reaction coordinate.