1. The small CRL4 CSA ubiquitin ligase component DDA1 regulates transcription-coupled repair dynamics.
- Author
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Llerena Schiffmacher DA, Lee SH, Kliza KW, Theil AF, Akita M, Helfricht A, Bezstarosti K, Gonzalo-Hansen C, van Attikum H, Verlaan-de Vries M, Vertegaal ACO, Hoeijmakers JHJ, Marteijn JA, Lans H, Demmers JAA, Vermeulen M, Sixma TK, Ogi T, Vermeulen W, and Pines A
- Subjects
- Humans, Cryoelectron Microscopy, Cullin Proteins metabolism, Cullin Proteins genetics, DNA Damage, HEK293 Cells, Protein Binding, Receptors, Interleukin-17, Transcription, Genetic, Ubiquitination, DNA-Binding Proteins metabolism, DNA-Binding Proteins genetics, Excision Repair, Ubiquitin-Protein Ligases metabolism, Ubiquitin-Protein Ligases genetics
- Abstract
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 CRL4CSA 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 CRL4CSA complex. Functional analysis reveals that DDA1 coordinates ubiquitination dynamics during TC-NER and is required for efficient turnover and progression of this process., (© 2024. The Author(s).)- Published
- 2024
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