Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair
Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair
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DOI：
10.1016/j.cell.2020.02.010
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发表时间：
2020-03-19
期刊：
影响因子：
64.5
通讯作者：
Ogi, Tomoo
中科院分区：
文献类型：
--
作者：
Nakazawa, Yuka;Hara, Yuichiro;Ogi, Tomoo
转录偶联核苷酸切除修复（TC-NER）是通过延长RNA聚合酶H（RNAPIIo）在DNA损伤处的停滞而启动的。响应DNA损伤的RNAPIIo的泛素化是进化上保守的事件，但其在哺乳动物中的功能尚不清楚。在这里，我们确定了一个单一的DNA损伤诱导的泛素化位点RNAPII在RPB 1-K1268，它调节转录恢复和DNA损伤抗性。从机制上讲，RPB 1-K1268泛素化通过也涉及UVSSA-K414泛素化的转移机制刺激核心-TFIIH复合物与停滞的RNAPIIo的缔合。我们开发了一种链特异性ChIP-seq方法，该方法揭示了RPB 1-K1268泛素化对于修复和解决DNA损伤处的转录瓶颈非常重要。最后，RPB 1-K1268 R基因敲入小鼠表现出寿命短、过早衰老和神经退行性变。我们的研究结果表明，RNAPII泛素化通过激活TC-NER和并行处理DNA损伤停滞的RNAPIIo提供了一种双重保护机制，它们共同防止了长时间的转录停滞并保护神经变性。
Transcription-coupled nucleotide excision repair (TC-NER) is initiated by the stalling of elongating RNA polymerase H (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.