Abstract

Transcription-coupled DNA repair (TCR) removes bulky DNA lesions that impede RNA polymerase II (RNAPII) transcription. TCR is initiated the stalling of RNAPII, and recent studies have outlined the stepwise assembly of TCR factors CSB, CSA, UVSSA, and TFIIH around the lesion-stalled polymerase. However, the mechanism and factors required for the transition to downstream repair steps, including RNAPII removal to provide repair proteins access to the DNA lesion, remain unclear. Here, we identify STK19 as a new TCR factor facilitating this transition. Loss of STK19 does not impact initial TCR complex assembly or RNAPII ubiquitylation but delays lesion-stalled RNAPII clearance, thereby interfering with the downstream repair reaction. Cryo-EM and mutational analysis reveal that STK19 associates with the TCR complex, positioning itself between RNAPII, UVSSA, and CSA. The structural insights and molecular modeling suggest that STK19 positions the ATPase subunits of TFIIH onto DNA in front of RNAPII. Together, these findings provide new insights into the factors and mechanisms required for TCR.

Biography

Martijn Luijsterburg received his M.Sc. in Medical Biology in 2004 from the Free University in Amsterdam. He received his Ph.D. in Cell Biology in 2008 from the University of Amsterdam during which he studied how DNA repair complexes involved in nucleotide excision repair assemble and function at sites of DNA damage in living cells (group of Prof. Roel van Driel). He became a postdoctoral fellow in the group of Prof. Nico Dantuma in 2008 at the Karolinska Institute in Stockholm for which he received long-term fellowships from the Netherlands Organization for Scientific Research (NWO-Rubicon) and the European Molecular Biology Organization (EMBO). His research at the Karolinska Institute focused on elucidating the role of ubiquitin ligases during DNA double-strand break repair and nucleotide excision repair. In 2011, he joined the group of Prof. Haico van Attikum at the Leiden University Medical Center (LUMC) for which he received long-term fellowships from the Federation of European Biochemical Societies (FEBS) and the Netherlands Organization for Scientific Research (NWO-VENI). His research in the Van Attikum group focused on elucidating the role of chromatin remodeling enzymes in the repair of DNA double-strand breaks.

In 2015, Martijn was appointed as group leader at the Department of Human Genetics of the Leiden University Medical Center for which he received an LUMC Research Fellowship and an NWO-VIDI grant. In 2017, he was appointed as tenured assistant professor. In 2021, Martijn was elected as EMBO young investigator. In 2022, Martijn received an ERC consolidator grant and NWO-VICI grant and was promoted to associate professor. His group investigates mechanisms in transcription-coupled DNA repair using genome-wide genetics, genomics and proteomic approaches. The goal of the lab is to increase our fundamental knowledge about how cells deal with obstacles caused by DNA damage during transcription, and to understand how – if this process fails – this can cause human diseases. Martijn is advisor for the Cockayne syndrome support group Amy and Friends in the UK (https://www.amyandfriends.org/Meet-the-Team) and Riaan Research Initiative in the US (https://riaanresearch.org/about-us/).