8QH5

CryoEM structure of UVSSA(VHS)-CSA-DDB1-DDA1

Summary for 8QH5

• Entry DOI: 10.2210/pdb8qh5/pdb
• EMDB information: 18398
• Descriptor: UV-stimulated scaffold protein A, DNA excision repair protein ERCC-8, DNA damage-binding protein 1, ... (4 entities in total)
• Functional Keywords: ubiquitin ligase, dna repair, ligase
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 274685.26

Authors

Lee, S.-H.,Sixma, T.K. (deposition date: 2023-09-06, release date: 2024-08-14, Last modification date: 2025-08-27)

Primary citation

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.
The small CRL4 CSA ubiquitin ligase component DDA1 regulates transcription-coupled repair dynamics.
Nat Commun, 15:6374-6374, 2024

PubMed 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). Although ubiquitination of several TC-NER proteins by CRL4 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 complex. Functional analysis reveals that DDA1 coordinates ubiquitination dynamics during TC-NER and is required for efficient turnover and progression of this process.

PubMed: 39075067
DOI: 10.1038/s41467-024-50584-7

Experimental method

ELECTRON MICROSCOPY (3.4 Å)