Polθ promotes the repair of 5'-DNA-protein crosslinks by microhomology-mediated end-joining.
Polθ promotes the repair of 5'-DNA-protein crosslinks by microhomology-mediated end-joining.
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DOI：
10.1016/j.celrep.2021.108820
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发表时间：
2021-03-09
期刊：
影响因子：
8.8
通讯作者：
Pomerantz RT
中科院分区：
文献类型：
--
作者：
Chandramouly G;Liao S;Rusanov T;Borisonnik N;Calbert ML;Kent T;Sullivan-Reed K;Vekariya U;Kashkina E;Skorski T;Yan H;Pomerantz RT
DNA聚合酶θ（Polθ）可使化疗药物（如拓扑异构酶抑制剂）对双链断裂（DSB）处的DNA-蛋白质交联（DPC）产生耐药性。这表明Polθ可能通过微同源介导的末端连接（MMEJ）促进DPC修复。在这里，我们调查Polθ修复DSB携带DPC监测MMEJ爪蟾卵提取物。提取物中的MMEJ依赖于Polθ，表现出MMEJ修复特征，并且独立于非同源末端连接和复制体有效地修复5′末端DPC。我们通过使用MMEJ报告基因证明了Polθ促进哺乳动物细胞5′端DPC的修复，并发现Polθ除了赋予拓扑异构酶抑制剂外，还赋予对甲醛的抗性。Polθ和酪氨酰-DNA磷酸二酯酶2（TDP 2）的双重缺陷导致细胞对依托泊苷的严重敏感性，这表明MMEJ是一种独立的DPC修复途径。这些研究概括了体外MMEJ，并阐明了Polθ如何赋予依托泊苷耐药性。Chandramouly等人发现Polθ保护细胞免受DNA-蛋白质交联剂（DPC）的影响，并促进非洲爪蟾卵提取物以及哺乳动物细胞中发生在双链断裂（DSB）处的DPC的微同源介导的末端连接（MMEJ）修复。Polθ介导的DPC修复不依赖于非同源末端连接（NHEJ）和同源重组（HR）。
DNA polymerase θ (Polθ) confers resistance to chemotherapy agents that cause DNA-protein crosslinks (DPCs) at double-strand breaks (DSBs), such as topoisomerase inhibitors. This suggests Polθ might facilitate DPC repair by microhomology-mediated end-joining (MMEJ). Here, we investigate Polθ repair of DSBs carrying DPCs by monitoring MMEJ in Xenopus egg extracts. MMEJ in extracts is dependent on Polθ, exhibits the MMEJ repair signature, and efficiently repairs 5′ terminal DPCs independently of non-homologous end-joining and the replisome. We demonstrate that Polθ promotes the repair of 5′ terminal DPCs in mammalian cells by using an MMEJ reporter and find that Polθ confers resistance to formaldehyde in addition to topoisomerase inhibitors. Dual deficiency in Polθ and tyrosyl-DNA phosphodiesterase 2 (TDP2) causes severe cellular sensitivity to etoposide, which demonstrates MMEJ as an independent DPC repair pathway. These studies recapitulate MMEJ in vitro and elucidate how Polθ confers resistance to etoposide. Chandramouly et al. find that Polθ protects cells from DNA-protein crosslink (DPC) agents and promotes microhomology-mediated end-joining (MMEJ) repair of DPCs occurring at double-strand breaks (DSBs) in Xenopus egg extracts as well as mammalian cells. Polθ-mediated repair of DPCs occurring at DSBs is independent of non-homologous end-joining (NHEJ) and homologous recombination (HR).
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影响因子：
4.5
作者：
Gómez-Herreros F;Romero-Granados R;Zeng Z;Alvarez-Quilón A;Quintero C;Ju L;Umans L;Vermeire L;Huylebroeck D;Caldecott KW;Cortés-Ledesma F
通讯作者：
Cortés-Ledesma F
影响因子：
14.9
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Liao S;Tammaro M;Yan H
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Yan H
影响因子：
64.8
作者：
Mateos-Gomez PA;Gong F;Nair N;Miller KM;Lazzerini-Denchi E;Sfeir A
通讯作者：
Sfeir A
影响因子：
14.9
作者：
Ahrabi S;Sarkar S;Pfister SX;Pirovano G;Higgins GS;Porter AC;Humphrey TC
通讯作者：
Humphrey TC
影响因子：
16.8
作者：
Kent, Tatiana;Chandramouly, Gurushankar;McDevitt, Shane Michael;Ozdemir, Ahmet Y.;Pomerantz, Richard T.
通讯作者：
Pomerantz, Richard T.