Journal Article

The small CRL4CSA ubiquitin ligase component DDA1 regulates transcription-coupled repair dynamics

Haico van Attikum, Angela Helfricht, Masaki Akita, Karel Bezstarosti, Camila Gonzalo-Hansen, Alfred C. O. Vertegaal, Titia K. Sixma, Wim Vermeulen, Jan H. J. Hoeijmakers, Arjan F. Theil, Tomoo Ogi, Jurgen A. Marteijn, Michiel Vermeulen, Matty Verlaan-de Vries, Jeroen A. A. Demmers, Katarzyna W. Kliza, Hannes Lans, Alex Pines, Diana A. Llerena Schiffmacher, Shun-Hsiao Lee

2024

Overview

Transcription-blocking DNA lesions are specifically targeted by transcription-coupled nucleotide excision repair (TC-NER), which removes a broad spectrum of DNA lesions to preserve transcriptional output and thereby cellular homeostasis to counteract aging. TC-NER is initiated by the stalling of RNA polymerase II at DNA lesions, which triggers the assembly of the TC-NER-specific proteins CSA, CSB and UVSSA. CSA, a WD40-repeat containing protein, is the substrate receptor subunit of a cullin-RING ubiquitin ligase complex composed of DDB1, CUL4A/B and RBX1 (CRL4 CSA ). Although ubiquitination of several TC-NER proteins by CRL4 CSA has been reported, it is still unknown how this complex is regulated. To unravel the dynamic molecular interactions and the regulation of this complex, we apply a single-step protein-complex isolation coupled to mass spectrometry analysis and identified DDA1 as a CSA interacting protein. Cryo-EM analysis shows that DDA1 is an integral component of the CRL4 CSA complex. Functional analysis reveals that DDA1 coordinates ubiquitination dynamics during TC-NER and is required for efficient turnover and progression of this process. Transcription-Coupled Nucleotide Excision Repair (TC-NER) removes transcription-blocking DNA lesions. This study reveals that DDA1 is a crucial player in TC-NER, protecting transcription programs against genotoxic insults.

• Publisher: Springer Science and Business Media LLC,Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 101/28 / 13/109 / 13/89 / 14/19 / 14/34 / 14/35 / 38/22 / 38/91 / 631/337/1427/1430 / 631/337/1427/2566 / 82/58 / Cullin / Deoxyribonucleic acid / DNA / DNA repair / DNA-directed RNA polymerase / Functional analysis / Genotoxicity / Homeostasis / Humanities and Social Sciences / Lesions / Mass spectrometry / Mass spectroscopy / Molecular Biology / Molecular interactions / multidisciplinary / Nucleotide excision repair / Nucleotides / Proteins / Proteomics and Chromatin Biology / Q / RNA polymerase / RNA polymerase II / Science / Science (multidisciplinary) / Substrates / Transcription-coupled repair / Ubiquitin-protein ligase / Ubiquitination