Cited by 19DNA Repair MechanismsUbiquitin and proteasome pathwaysGenomics and Chromatin Dynamics

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

Open access
Llerena Schiffmacher, Diana A., Lee, Shun-Hsiao, Kliza, Katarzyna W., Theil, Arjan F., Akita, Masaki, Helfricht, Angela, Bezstarosti, Karel, Gonzalo-Hansen, Camila +12 more·2024·Nature Communications

Published version of a preprint

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 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.

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Authors
Llerena Schiffmacher, D. A., Lee, S.-H., Kliza, K. W., Theil, A. F., Akita, M., Helfricht, A., Bezstarosti, K., Gonzalo-Hansen, C., van Attikum, H., Verlaan-de Vries, M., Vertegaal, A. C. O., Hoeijmakers, J. H. J., Marteijn, J. A., Lans, H., Demmers, J. A. A., Vermeulen, M., Sixma, T. K., Ogi, T., Vermeulen, W., & Pines, A.
Affiliations
  • The Netherlands Cancer Institute (NL)
  • Radboud University Nijmegen (NL)
  • Erasmus MC (NL)
  • National University of Singapore (SG)
  • Leiden University Medical Center (NL)
  • Research Institute of Environmental Medicine, Nagoya University (JP)
Corresponding author
  • Vermeulen, Wim
  • Pines, Alex
Venue
Nature Communications
Published
2024-07-29
Volume
15
Issue
1
Pages
6374-6374
ISSN
2041-1723 (ISSN-L)
OpenAlex
W4401084554
Cited by
19
References
98
Citations by year
2026: 6 · 2025: 10 · 2024: 3
FWCI
2.59
Type
journal article
Language
en
Open access
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • EUR Research Repository (Erasmus University Rotterdam) (publishedVersion, cc-by)
  • Leiden Repository (Leiden University) (submittedVersion, cc-by)
  • Radboud Repository (Radboud University) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • EUR Research Repository (Erasmus University Rotterdam) (submittedVersion, cc-by)
Funders
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (714.017.003)
  • European Molecular Biology Organization (ALTF 1159-2019)
  • U.S. Department of Health & Human Services | NIH | National Institute on Aging (P01 AG017242)
  • ZonMw (733050810)
  • Deutsche Forschungsgemeinschaft (496650118)
  • European Joint Project on Rare Diseases RD20-113, acronym TC-NER
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
transcription-coupled repair, CSA, RNA polymerase II stalling, protein complex formation, WD40 repeat proteins, DNA repair regulation

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