TRiC controls transcription resumption after UV damage by regulating Cockayne syndrome protein A.

Abstract

Transcription-blocking DNA lesions are removed by transcription-coupled nucleotide excision repair (TC-NER) to preserve cell viability. TC-NER is triggered by the stalling of RNA polymerase II at DNA lesions, leading to the recruitment of TC-NER-specific factors such as the CSA-DDB1-CUL4A-RBX1 cullin-RING ubiquitin ligase complex (CRLCSA). Despite its vital role in TC-NER, little is known about the regulation of the CRLCSA complex during TC-NER. Using conventional and cross-linking immunoprecipitations coupled to mass spectrometry, we uncover a stable interaction between CSA and the TRiC chaperonin. TRiC's binding to CSA ensures its stability and DDB1-dependent assembly into the CRLCSA complex. Consequently, loss of TRiC leads to mislocalization and depletion of CSA, as well as impaired transcription recovery following UV damage, suggesting defects in TC-NER. Furthermore, Cockayne syndrome (CS)-causing mutations in CSA lead to increased TRiC binding and a failure to compose the CRLCSA complex. Thus, we uncover CSA as a TRiC substrate and reveal that TRiC regulates CSA-dependent TC-NER and the development of CS.

• Authors: Alex Pines, Madelon Dijk, Matthew Makowski, Elisabeth M Meulenbroek, Mischa G Vrouwe, Yana van der Weegen, Marijke Baltissen, Pim J French, Martin E van Royen, Martijn S Luijsterburg, Leon H Mullenders, Michiel Vermeulen, Wim Vermeulen, Navraj S Pannu, Haico van Attikum
• Journal: Nature communications (Nat Commun) Vol. 9 Issue 1 Pg. 1040 (03 12 2018) ISSN: 2041-1723 [Electronic] England
• PMID: 29531219 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ERCC8 protein, human
• Transcription Factors
• Chaperonin Containing TCP-1
• DNA Repair Enzymes

Topics

• Blotting, Western
• Cell Line, Tumor
• Cell Survival (genetics, radiation effects)
• Chaperonin Containing TCP-1 (genetics, metabolism)
• Cockayne Syndrome (genetics)
• DNA Damage (genetics, radiation effects)
• DNA Repair Enzymes (genetics, metabolism)
• Humans
• Immunoprecipitation
• Mass Spectrometry
• Microscopy, Fluorescence
• Mutation (genetics)
• RNA Interference
• Transcription Factors (genetics, metabolism)
• Transcription, Genetic (genetics, radiation effects)
• Ultraviolet Rays