The cooperative action of CSB, CSA, and UVSSA target TFIIH to DNA damage-stalled RNA polymerase II

• Published in: Nature communications 2020-04, Vol.11 (1), p.2104-2104, Article 2104
• Main Authors: van der Weegen, Yana, Golan-Berman, Hadar, Mevissen, Tycho E. T., Apelt, Katja, González-Prieto, Román, Goedhart, Joachim, Heilbrun, Elisheva E., Vertegaal, Alfred C. O., van den Heuvel, Diana, Walter, Johannes C., Adar, Sheera, Luijsterburg, Martijn S.
• Format: Article
• Language: English
• Subjects: 13/106; 14/35; 45/15; 631/337/1427; 631/337/1427/1430; 631/337/572; 82/1; 82/51; 82/58; Animals; Assembly; Carrier Proteins - metabolism; Cell Line, Tumor; Damage; Deoxyribonucleic acid; DNA; DNA Damage; DNA Helicases - metabolism; DNA Repair; DNA Repair Enzymes - metabolism; DNA-directed RNA polymerase; Humanities and Social Sciences; Humans; multidisciplinary; Poly-ADP-Ribose Binding Proteins - metabolism; Proteins; Repair; Ribonucleic acid; RNA; RNA polymerase; RNA polymerase II; RNA Polymerase II - metabolism; Science; Science (multidisciplinary); Strands; Transcription Factor TFIIH - metabolism; Transcription Factors - metabolism; Transcription, Genetic; Transcription-coupled repair; Ultraviolet Rays; Xenopus laevis
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-15903-8

The response to DNA damage-stalled RNA polymerase II (RNAPIIo) involves the assembly of the transcription-coupled repair (TCR) complex on actively transcribed strands. The function of the TCR proteins CSB, CSA and UVSSA and the manner in which the core DNA repair complex, including transcription factor IIH (TFIIH), is recruited are largely unknown. Here, we define the assembly mechanism of the TCR complex in human isogenic knockout cells. We show that TCR is initiated by RNAPIIo-bound CSB, which recruits CSA through a newly identified CSA-interaction motif (CIM). Once recruited, CSA facilitates the association of UVSSA with stalled RNAPIIo. Importantly, we find that UVSSA is the key factor that recruits the TFIIH complex in a manner that is stimulated by CSB and CSA. Together these findings identify a sequential and highly cooperative assembly mechanism of TCR proteins and reveal the mechanism for TFIIH recruitment to DNA damage-stalled RNAPIIo to initiate repair. The response to DNA damage-stalled RNA polymerase II leads to the assembly of the transcription-coupled repair (TCR) complex on actively transcribed strands. Here, the authors reveal the complex assembly mechanism of the TCR complex in human cells.