In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4^CSA, which cooperate with UVSSA and ELOF1 to recruit TFIIH. To explore the mechanism of TC-NER, we recapitulated this reaction in vitro. When a plasmid containing a site-specific lesion is transcribed in frog egg extract, error-free repair is observed that depends on CSB, CRL4^CSA, UVSSA, and ELOF1. Repair also requires STK19, a factor previously implicated in transcription recovery after UV exposure. A 1.9-Å cryo-electron microscopy structure shows that STK19 binds the TC-NER complex through CSA and the RPB1 subunit of RNA Pol II. Furthermore, AlphaFold predicts that STK19 interacts with the XPD subunit of TFIIH, and disrupting this interface impairs cell-free repair. Molecular modeling suggests that STK19 positions TFIIH ahead of RNA Pol II for lesion verification. Our analysis of cell-free TC-NER suggests that STK19 couples RNA Pol II stalling to downstream repair events.

中文翻译：

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STK19 定位 TFIIH 进行游离转录偶联 DNA 修复


在转录偶联核苷酸切除修复 （TC-NER） 中，停滞的 RNA 聚合酶 II （RNA Pol II） 结合 CSB 和 CRL4^CSA，它们与 UVSSA 和 ELOF1 合作募集 TFIIH。为了探索 TC-NER 的机制，我们在体外概括了这个反应 。当在青蛙卵提取物中转录含有位点特异性损伤的质粒时，观察到依赖于 CSB、CRL4^CSA、UVSSA 和 ELOF1 的无错误修复。修复还需要 STK19，这是以前与紫外线暴露后转录恢复有关的一个因素。1.9 Å 冷冻电子显微镜结构显示，STK19 通过 CSA 和 RNA Pol II 的 RPB1 亚基结合 TC-NER 复合物。此外，AlphaFold 预测 STK19 与 TFIIH 的 XPD 亚基相互作用，破坏该界面会损害无细胞修复。分子建模表明，STK19 将 TFIIH 置于 RNA Pol II 之前以进行病变验证。我们对游离 TC-NER 的分析表明，STK19 将 RNA Pol II 停滞与下游修复事件偶联。