@article{d86bdf9056724d55a4821d888e5f74ee,
title = "Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair",
abstract = "Transcription-coupled nucleotide excision repair (TC-NER) is initiated by the stalling of elongating RNA polymerase II (RNAPIIo) at DNA lesions. The ubiquitination of RNAPIIo in response to DNA damage is an evolutionarily conserved event, but its function in mammals is unknown. Here, we identified a single DNA damage-induced ubiquitination site in RNAPII at RPB1-K1268, which regulates transcription recovery and DNA damage resistance. Mechanistically, RPB1-K1268 ubiquitination stimulates the association of the core-TFIIH complex with stalled RNAPIIo through a transfer mechanism that also involves UVSSA-K414 ubiquitination. We developed a strand-specific ChIP-seq method, which revealed RPB1-K1268 ubiquitination is important for repair and the resolution of transcriptional bottlenecks at DNA lesions. Finally, RPB1-K1268R knockin mice displayed a short life-span, premature aging, and neurodegeneration. Our results reveal RNAPII ubiquitination provides a two-tier protection mechanism by activating TC-NER and, in parallel, the processing of DNA damage-stalled RNAPIIo, which together prevent prolonged transcription arrest and protect against neurodegeneration.",
keywords = "CRL/CSA/CSB, CS, ChIP-seq, Cockayne syndrome, NER, RNA polymerase II, RNAPII, TCR, TFIIH, UV-sensitive syndrome, UVSSA, UVsS, nucleotide excision repair, transcription coupled repair, ubiquitination of RNAPII",
author = "Yuka Nakazawa and Yuichiro Hara and Yasuyoshi Oka and Okiru Komine and {van den Heuvel}, Diana and Chaowan Guo and Yasukazu Daigaku and Mayu Isono and Yuxi He and Mayuko Shimada and Kana Kato and Nan Jia and Satoru Hashimoto and Yuko Kotani and Yuka Miyoshi and Miyako Tanaka and Akira Sobue and Norisato Mitsutake and Takayoshi Suganami and Akio Masuda and Kinji Ohno and Shinichiro Nakada and Tomoji Mashimo and Koji Yamanaka and Luijsterburg, {Martijn S.} and Tomoo Ogi",
note = "Funding Information: This paper is dedicated to the memory of a wonderful lady, Amy Garton-Hughes, who made a huge difference in the lives of Cockayne syndrome children. We are grateful to Drs. Alan Lehman and Chikahide Masutani for their helpful comments and discussions on the manuscript. We are grateful to Dr. Masato Kanemaki for his advice on the CRISPR/Cas9-based gene targeting experiments. Bioinformatics data processing was partially performed using the super-computing resource provided by Human Genome Center, the Institute of Medical Science, the University of Tokyo. This work was supported by the Special Coordination Funds for Rare and Intractable Diseases from Japan Agency for Medical Research and Development (AMED) (JP19ek0109281, JP19ek0109229, and JP19ek0109301), Grants in Aid for Scientific Research KAKENHI (JP15H02654 and JP17H00783) from the Japan Society for the Promotion of Science, a Science Research Grant from the Uehara Memorial Foundation, a grant from the Daiko Foundation, and a medical research grant from the Takeda Science Foundation to T.O.; KAKENHI (JP17H01877) and a medical research grant from Takeda Science Foundation to Y.N.; TBRF Postdoctoral Fellowship for Asian Researchers in Japan from The Tokyo Biochemical Research Foundation (TBRF) to C.G. and T.O.; and an LUMC research fellowship and an NWO-VIDI grant (016.161.320) to M.S.L. T.O. designed the study and experiments. Y.N. Y.O. D.V.d.H. C.G. Y.D. M.I. Y. He, M.S. K.K. N.J. S.H. M.S.L, and T.O. performed molecular and cell biological experiments. Y. Hara, and T.O. performed bioinformatics analyses. Y.O. O.K. C.G. Y. He, M.S. Y.K. Y.M. M.T. A.S. T.M. K.Y. and T.O. performed animal studies. N.M and S.N. contributed materials. N.M. T.S. A.M. K.O. S.N. T.M. K.Y. M.S.L. and T.O. coordinated the study. M.S.L. and T.O. wrote the manuscript. All authors commented on the manuscript. The authors declare no competing interests. Funding Information: This paper is dedicated to the memory of a wonderful lady, Amy Garton-Hughes, who made a huge difference in the lives of Cockayne syndrome children. We are grateful to Drs. Alan Lehman and Chikahide Masutani for their helpful comments and discussions on the manuscript. We are grateful to Dr. Masato Kanemaki for his advice on the CRISPR/Cas9-based gene targeting experiments. Bioinformatics data processing was partially performed using the super-computing resource provided by Human Genome Center, the Institute of Medical Science, the University of Tokyo. This work was supported by the Special Coordination Funds for Rare and Intractable Diseases from Japan Agency for Medical Research and Development (AMED) ( JP19ek0109281 , JP19ek0109229 , and JP19ek0109301 ), Grants in Aid for Scientific Research KAKENHI ( JP15H02654 and JP17H00783 ) from the Japan Society for the Promotion of Science , a Science Research Grant from the Uehara Memorial Foundation , a grant from the Daiko Foundation , and a medical research grant from the Takeda Science Foundation to T.O.; KAKENHI ( JP17H01877 ) and a medical research grant from Takeda Science Foundation to Y.N.; TBRF Postdoctoral Fellowship for Asian Researchers in Japan from The Tokyo Biochemical Research Foundation (TBRF) to C.G. and T.O.; and an LUMC research fellowship and an NWO-VIDI grant ( 016.161.320 ) to M.S.L. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = mar,
day = "19",
doi = "10.1016/j.cell.2020.02.010",
language = "English",
volume = "180",
pages = "1228--1244.e24",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "6",
}