A scanning-to-incision switch in TFIIH-XPG induced by DNA damage licenses nucleotide excision repair.
A scanning-to-incision switch in TFIIH-XPG induced by DNA damage licenses nucleotide excision repair.
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DOI：
10.1093/nar/gkac1095
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发表时间：
2023-02-22
影响因子：
14.9
通讯作者：
Hamdan, Samir M.
中科院分区：
文献类型：
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作者：
Bralic, Amer;Tehseen, Muhammad;Sobhy, Mohamed A.;Tsai, Chi-Lin;Alhudhali, Lubna;Yi, Gang;Yu, Jina;Yan, Chunli;Ivanov, Ivaylo;Tsutakawa, Susan E.;Tainer, John A.;Hamdan, Samir M.
核苷酸切除修复(NER)对于去除巨大的DNA碱基损伤和避免疾病至关重要。NER通过XPC识别病变，通过XPB和XPD ATPase分离链，然后通过XPF和XPG核酸酶切除病变。在这里，我们描述了XPG的主要调控机制和作用，以及它的切割活性之外的作用。值得注意的是，通过结合单分子成像和整体切割分析，我们发现XPG与7亚单位TFIIH核心(CoreTFIIH)的结合可刺激核心TFIIH依赖的双链(DS)DNA解开10倍，并刺激依赖XPG的DNA切割高达700倍。同时监测核心TFIIH单链(Ss)DNA易位和dsDNA解离的速率表明，XPG通过将ssDNA易位转换为dsDNA解离作为可能的承诺步骤来发挥作用。与NER途径调节有关，XPG切割活性在核心TFIIH在DNA上的移位过程中受到抑制，但当核心TFIIH在病变处停滞或当ATP水解酶被阻止时，XPG切割活性被许可。此外，≥的15个核苷酸的5‘-单链DNA是有效的移位和切割的先决条件。我们的结果揭示了一种配对的协调机制，在该机制中，关键的病变扫描和DNA切割步骤被顺序地协调，并且只有在产生≥15个核苷酸的5‘-单链DNA之后才允许去除受损的斑块，从而确保在切割之前正确的单链DNA气泡大小。ATP15核苷酸5‘单链DNA的拉伸允许适当的核心TFIIH-XPG复合体的协调，在≥存在的情况下有效地解离，但在没有或遇到损伤时裂解。
Nucleotide excision repair (NER) is critical for removing bulky DNA base lesions and avoiding diseases. NER couples lesion recognition by XPC to strand separation by XPB and XPD ATPases, followed by lesion excision by XPF and XPG nucleases. Here, we describe key regulatory mechanisms and roles of XPG for and beyond its cleavage activity. Strikingly, by combing single-molecule imaging and bulk cleavage assays, we found that XPG binding to the 7-subunit TFIIH core (coreTFIIH) stimulates coreTFIIH-dependent double-strand (ds)DNA unwinding 10-fold, and XPG-dependent DNA cleavage by up to 700-fold. Simultaneous monitoring of rates for coreTFIIH single-stranded (ss)DNA translocation and dsDNA unwinding showed XPG acts by switching ssDNA translocation to dsDNA unwinding as a likely committed step. Pertinent to the NER pathway regulation, XPG incision activity is suppressed during coreTFIIH translocation on DNA but is licensed when coreTFIIH stalls at the lesion or when ATP hydrolysis is blocked. Moreover, ≥15 nucleotides of 5′-ssDNA is a prerequisite for efficient translocation and incision. Our results unveil a paired coordination mechanism in which key lesion scanning and DNA incision steps are sequentially coordinated, and damaged patch removal is only licensed after generation of ≥15 nucleotides of 5′-ssDNA, ensuring the correct ssDNA bubble size before cleavage. A ≥15 nucleotide 5′ ssDNA stretch allows for proper coreTFIIH-XPG complex coordination that efficiently unwinds in the presence of ATP but cleaves in its absence or upon damage encounter.
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影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
14.9
作者：
Fu, Iwen;Mu, Hong;Geacintov, Nicholas E.;Broyde, Suse
通讯作者：
Broyde, Suse
影响因子：
--
作者：
Finger, L David;Atack, John M;Shen, Binghui
通讯作者：
Shen, Binghui
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
DOI：
10.1083/jcb.201006011
发表时间：
2011-02-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Overmeer RM;Moser J;Volker M;Kool H;Tomkinson AE;van Zeeland AA;Mullenders LH;Fousteri M
通讯作者：
Fousteri M