Journal Article

Transcription-coupled DNA-protein crosslink repair by CSB and CRL4 CSA -mediated degradation

Thijssen, Karen L, Lans, Hannes, Somsen, Hedda B, Dekkers, Dick H W, Demmers, Joris J P G, van Toorn, Marvin, Wienecke, Hannah Lena, Raams, Anja, Häckes, David, Gonzalo-Hansen, Camila, Llerena Schiffmacher, Diana A, Yu, Qing, Demmers, Jeroen A A, Lammers, Judith G, Ramadhin, Anisha R, Zhou, Di, Pines, Alex, Marteijn, Jurgen A, Janssens, Roel C, van der Woude, Melanie, van den Berg, Debbie L C, van Sluis, Marjolein, Vermeulen, Wim, Davó-Martínez, Carlota, Dealy, Shannon C, Pothof, Joris

2024

Overview

DNA-protein crosslinks (DPCs) arise from enzymatic intermediates, metabolism or chemicals like chemotherapeutics. DPCs are highly cytotoxic as they impede DNA-based processes such as replication, which is counteracted through proteolysis-mediated DPC removal by spartan (SPRTN) or the proteasome. However, whether DPCs affect transcription and how transcription-blocking DPCs are repaired remains largely unknown. Here we show that DPCs severely impede RNA polymerase II-mediated transcription and are preferentially repaired in active genes by transcription-coupled DPC (TC-DPC) repair. TC-DPC repair is initiated by recruiting the transcription-coupled nucleotide excision repair (TC-NER) factors CSB and CSA to DPC-stalled RNA polymerase II. CSA and CSB are indispensable for TC-DPC repair; however, the downstream TC-NER factors UVSSA and XPA are not, a result indicative of a non-canonical TC-NER mechanism. TC-DPC repair functions independently of SPRTN but is mediated by the ubiquitin ligase CRL4 and the proteasome. Thus, DPCs in genes are preferentially repaired in a transcription-coupled manner to facilitate unperturbed transcription.

• Subject: Carrier Proteins / DNA - genetics / DNA - metabolism / DNA Damage / DNA Helicases - genetics / DNA Helicases - metabolism / DNA Repair / DNA Repair Enzymes - genetics / DNA Repair Enzymes - metabolism / DNA-Binding Proteins - genetics / DNA-Binding Proteins - metabolism / HEK293 Cells / Humans / Poly-ADP-Ribose Binding Proteins - genetics / Poly-ADP-Ribose Binding Proteins - metabolism / Proteasome Endopeptidase Complex - metabolism / Proteolysis / Receptors, Interleukin-17 / RNA Polymerase II - genetics / RNA Polymerase II - metabolism / Transcription Factors - genetics / Transcription Factors - metabolism / Transcription, Genetic / Ubiquitin-Protein Ligases - genetics / Ubiquitin-Protein Ligases - metabolism