UV-sensitive syndrome protein UVSSA recruits USP7 to regulate transcription-coupled repair

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Abstract

Transcription-coupled nucleotide-excision repair (TC-NER) is a subpathway of NER that efficiently removes the highly toxic RNA polymerase II blocking lesions in DNA. Defective TC-NER gives rise to the human disorders Cockayne syndrome and UV-sensitive syndrome (UVSS)1. NER initiating factors are known to be regulated by ubiquitination2. Using a SILAC-based proteomic approach, we identified UVSSA (formerly known as KIAA1530) as part of a UV-induced ubiquitinated protein complex. Knockdown of UVSSA resulted in TC-NER deficiency. UVSSA was found to be the causative gene for UVSS, an unresolved NER deficiency disorder3. The UVSSA protein interacts with elongating RNA polymerase II, localizes specifically to UV-induced lesions, resides in chromatin-associated TC-NER complexes and is implicated in stabilizing the TC-NER master organizing protein ERCC6 (also known as CSB) by delivering the deubiquitinating enzyme USP7 to TC-NER complexes. Together, these findings indicate that UVSSA-USP7–mediated stabilization of ERCC6 represents a critical regulatory mechanism of TC-NER in restoring gene expression.

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Figure 1: UVSSA knockdown results in reduced TC-NER activity.
Figure 2: UVSSA expression rescues TC-NER deficiency in UVSS-A (TA-24) cells.
Figure 3: UVSSA is recruited to active TC-NER sites in a UV damage– and transcription-dependent manner.
Figure 4: UVSSA-dependent recruitment of USP7 to active TC-NER complexes stabilizes ERCC6 and restores UV-inhibited RNA synthesis.

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Acknowledgements

We thank R. Bernards and M. Epping (Nederlands Kanker Instituut) for the Myc-tagged USP7 expression construct and P. Verrijzer and A. Reddy (Erasmus Medical Centre) for shUSP7-expressing lentivirus. We thank H. Slor (Tel Aviv University) for the TA-24sv40 cell line and N.G.J. Jaspers and H. Lans for discussions and critical reading of the manuscript. This work was funded by the Netherlands Genomics Initiative NPCII (to P.S.), 935.19.021 and 935.11.042 (to W.V., C.L. and J.A.M.), the Dutch Organization for Scientific Research ZonMW Veni Grant (917.96.120 to J.A.M.) and TOP grant (912.08.031 to W.V.), Marie Curie FP7-PIEF-GA-2009-253544 (to M.F.), the Association for International Cancer Research (10-594 to W.V.) and the Cancer Genomics Centre and ERC (advanced research grant to J.H.J.H.).

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D.H.W.D. and J.A.A.D. performed the mass spectrometry analyses, A.R. performed UDS and RRS experiments, A.C.v.d.H. performed immunoprecipitation experiments, C.L. provided technical assistance, and M.F. designed and, together with A.L., performed ChIP experiments. J.H.J.H. provided support and advice, and helped write the manuscript. P.S. and J.A.M. performed experiments and generated reagents. W.V. and J.A.M. designed the study and supervised the project. W.V., J.A.M. and P.S. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Wim Vermeulen or Jurgen A Marteijn.

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The authors declare no competing financial interests.

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Schwertman, P., Lagarou, A., Dekkers, D. et al. UV-sensitive syndrome protein UVSSA recruits USP7 to regulate transcription-coupled repair. Nat Genet 44, 598–602 (2012). https://doi.org/10.1038/ng.2230

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