DNA Repair最新文献

英文中文

Functions of PMS2 and MLH1 important for regulation of divergent repeat-mediated deletions

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-26 DOI: 10.1016/j.dnarep.2024.103791

Hannah Trost , Felicia Wednesday Lopezcolorado , Arianna Merkell , Jeremy M. Stark

Repeat-mediated deletions (RMDs) are a type of deletion rearrangement that utilizes two repetitive elements to bridge a DNA double-strand break (DSB) that leads to loss of the intervening sequence and one of the repeats. Sequence divergence between repeats causes RMD suppression and indeed this divergence must be resolved in the RMD products. The mismatch repair factor, MLH1, was shown to be critical for both RMD suppression and a polarity of sequence divergence resolution in RMDs. Here, we sought to study the interrelationship between these two aspects of RMD regulation (i.e., RMD suppression and polar divergence resolution), by examining several mutants of MLH1 and its binding partner PMS2. To begin with, we show that PMS2 is also critical for both RMD suppression and polar resolution of sequence divergence in RMD products. Then, with six mutants of the MLH1-PMS2 heterodimer, we found several different patterns: three mutants showed defects in both functions, one mutant showed loss of RMD suppression but not polar divergence resolution, whereas another mutant showed the opposite, and finally one mutant showed loss of RMD suppression but had a complex effect on polar divergence resolution. These findings indicate that RMD suppression vs. polar resolution of sequence divergence are distinct functions of MLH1-PMS2.

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引用次数: 0

Why the ROS matters: One-electron oxidants focus DNA damage and repair on G-quadruplexes for gene regulation ROS 为何重要？单电子氧化剂使 DNA 损伤和修复集中于 G 型四联体，从而实现基因调控

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-16 DOI: 10.1016/j.dnarep.2024.103789

Aaron M. Fleming, Cynthia J. Burrows

Hydrogen peroxide is a precursor to reactive oxygen species (ROS) in cells because of its high reactivity with iron(II) carbonate complexes formed in the labile iron pool due to a high concentration of intracellular bicarbonate (25–100 mM). This chemistry leads to the formation of carbonate radical anion rather than hydroxyl radical, and unlike the latter ROS, CO₃^•- is a milder one-electron oxidant with high specificity for guanine oxidation in DNA and RNA. In addition to metabolism, another major source of DNA oxidation is inflammation which generates peroxynitrite, another precursor to CO₃^•- via reaction with dissolved CO₂. The identity of the ROS is important because not all radicals react with DNA in the same way. Whereas hydroxyl radical forms adducts at all four bases and reacts with multiple positions on ribose leading to base loss and strand breaks, carbonate radical anion is focused on guanosine oxidation to yield 8-oxo-7,8-dihydroguanosine in nucleic acids and the nucleotide pool, a modification that can function epigenetically in the context of a G-quadruplex. DNA sequences of multiple adjacent guanines, as found in G-quadruplex-forming sequences of gene promoters, are particularly susceptible to oxidative damage, and the focusing of CO₃^•- chemistry on these sites can lead to a transcriptional response during base excision repair. In this pathway, AP-endonuclease 1 plays a key role in accelerating G-quadruplex folding as well as recruiting activating transcription factors to impact gene expression.

过氧化氢是细胞中活性氧（ROS）的前体，因为它与细胞内高浓度碳酸氢盐（25-100 毫摩尔）在易失铁池中形成的碳酸铁（II）络合物具有高度反应性。这种化学反应会形成碳酸根阴离子而不是羟基自由基，与羟基自由基不同，CO3--是一种较温和的单电子氧化剂，对 DNA 和 RNA 中的鸟嘌呤氧化具有高度特异性。除新陈代谢外，DNA 氧化的另一个主要来源是炎症，炎症通过与溶解的 CO2 反应产生过亚硝酸盐，这是 CO3--的另一种前体。ROS 的特性非常重要，因为并非所有自由基都以相同的方式与 DNA 发生反应。羟基自由基会在所有四个碱基上形成加合物，并与核糖上的多个位置发生反应，导致碱基缺失和链断裂，而碳酸自由基阴离子则主要作用于鸟苷氧化，在核酸和核苷酸池中生成 8-氧代-7,8-二氢鸟苷，这种修饰可在 G 型四联体中发挥表观遗传作用。在基因启动子的 G-四叠体形成序列中发现的多个相邻鸟嘌呤的 DNA 序列特别容易受到氧化损伤，在碱基切除修复过程中，CO3-化学作用集中在这些位点上会导致转录反应。在这一途径中，AP-内切酶 1 在加速 G-四叠体折叠以及招募激活转录因子以影响基因表达方面起着关键作用。

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引用次数: 0

PrimPol-mediated repriming elicits gap-filling by template switching and promotes cellular tolerance to cidofovir PrimPol介导的斥责可通过模板切换引起间隙填充，并促进细胞对西多福韦的耐受性。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-14 DOI: 10.1016/j.dnarep.2024.103787

Mubasshir Washif, Ryotaro Kawasumi, Kouji Hirota

A nucleoside analog, Cidofovir (CDV), is used for the treatment of viral diseases such as cytomegalovirus retinitis and herpes virus infection. CDV converts to its active diphosphate metabolite (CDVpp) through cellular kinases and acts as a competitive inhibitor for viral polymerase thereby interfering with viral replication. However, the effect of this drug on the replication of healthy host cells and the mechanisms involved in the cellular tolerance to CDV are yet to be fully understood. In this study, we explored the mechanisms underlying cellular tolerance to CDV by screening mutant cell lines exhibiting hypersensitivity to CDV from a collection of DT40 mutants deficient in various genome maintenance systems. We identified Rad17 and PrimPol as critical factors for CDV tolerance. We found that Rad17 plays a pivotal role in activating intra-S phase checkpoint by the phosphorylation of Chk1, a vital checkpoint mediator. We showed that PrimPol, a factor involved in the release of stalled replication, plays critical roles in CDV tolerance in tandem with Rad17. We found that PrimPol deficient cells showed slower replication on the CDV-incorporated template strand than did wild-type cells, indicating a critical role of PrimPol in the continuous replication fork progression on the CDV-incorporated damaged template. PrimPol releases replication arrest with its DNA-damage bypass function and its repriming function, we thus investigated which PrimPol function is involved in CDV tolerance using the separation of function mutant genes of PRIMPOL. The CDV hypersensitive phenotype of PrimPol deficient cells was restored by PRIMPOL^Y89D (primase active / reduced polymerase activity), indicating that the repriming function of PrimPol is required for maintaining replication on the CDV-damaged template. Moreover, we found that the number of sister chromatid exchange (SCE) was reduced in PrimPol-deficient cells. These data indicate that gaps generated by PrimPol-mediated repriming on CDV-damaged templates promote post-replicative gap-filing by template switching.

核苷类似物西多福韦（CDV）用于治疗巨细胞病毒视网膜炎和疱疹病毒感染等病毒性疾病。CDV 通过细胞激酶转化为其活性二磷酸代谢物（CDVpp），成为病毒聚合酶的竞争性抑制剂，从而干扰病毒复制。然而，这种药物对健康宿主细胞复制的影响以及细胞对 CDV 的耐受机制尚待充分了解。在这项研究中，我们从一系列缺乏各种基因组维护系统的 DT40 突变体中筛选出了对 CDV 表现出超敏反应的突变细胞系，从而探索了细胞耐受 CDV 的机制。我们发现Rad17和PrimPol是CDV耐受性的关键因素。我们发现，Rad17 在通过磷酸化 Chk1（一种重要的检查点介导因子）激活 S 期内检查点方面起着关键作用。我们发现，参与释放停滞复制的因子PrimPol与Rad17一起在CDV耐受中发挥关键作用。我们发现，与野生型细胞相比，PrimPol 缺陷细胞在 CDV 导入的模板链上的复制速度更慢，这表明 PrimPol 在 CDV 导入的受损模板上的持续复制叉进展中起着关键作用。PrimPol通过其DNA损伤旁路功能和斥责功能释放复制停滞，因此我们利用PRIMPOL的分离功能突变基因研究了PrimPol的哪种功能参与了CDV耐受。PrimPol缺陷细胞的CDV超敏表型通过PRIMPOLY89D（引物酶活性/聚合酶活性降低）得以恢复，这表明PrimPol的斥责功能是在CDV损伤模板上维持复制所必需的。此外，我们还发现，PrimPol缺陷细胞中姐妹染色单体交换（SCE）的数量减少了。这些数据表明，PrimPol 在 CDV 破坏的模板上介导的修复所产生的间隙通过模板切换促进了复制后的间隙整理。

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引用次数: 0

DNAR special issue: DNA damage responses and neurological disease The DNA damage response and neurological disease DNAR 特刊：DNA 损伤反应与神经系统疾病 DNA 损伤反应与神经系统疾病。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-14 DOI: 10.1016/j.dnarep.2024.103788

Keith Caldecott

引用次数: 0

The interferon response at the intersection of genome integrity and innate immunity 基因组完整性与先天性免疫交汇处的干扰素反应。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-14 DOI: 10.1016/j.dnarep.2024.103786

Filip D. Duzanic, Lorenza Penengo

In recent years, numerous reports indicated that, besides pathogen infections, DNA replication stress and defective DNA repair can trigger the innate immune response by introducing a state of viral mimicry, due to cytosolic accumulation of the self-nucleic acid species, which culminates in the activation of type I interferon (IFN) pathway. In turn, IFN upregulates a variety of factors mutually implicated in immune- and genome-related mechanisms, shedding light on the unprecedented causality between genome stability and innate immunity. Intriguingly, in addition to being induced by replication stress, IFN-regulated factors can also promote it, pinpointing IFN signaling as both a consequence and a cause of replication stress. Here, we provide an overview of the factors and molecular mechanisms implicated in the evolutionary conserved crosstalk between genome maintenance and innate immunity, highlighting the role of the IFN-stimulated gene 15 (ISG15), which appears to be at the hub of this intersection. Moreover, we discuss the potential significance and clinical implications of the immune-mediated modulation of DNA replication and repair upon pathogen infection and in human diseases such as cancer and autoinflammatory syndromes. Finally, we discuss the relevant open questions and future directions.

近年来，大量报道表明，除了病原体感染外，DNA 复制应激和 DNA 修复缺陷可通过引入一种病毒模拟状态来触发先天性免疫反应，这是由于自身核酸种类在细胞膜上积累，最终激活了 I 型干扰素（IFN）通路。反过来，IFN 会上调与免疫和基因组相关机制相互牵连的各种因素，从而揭示基因组稳定性与先天免疫之间前所未有的因果关系。耐人寻味的是，IFN调控因子除了能被复制压力诱导外，还能促进复制压力，这表明 IFN 信号既是复制压力的结果，也是复制压力的原因。在此，我们概述了基因组维护与先天免疫之间进化保守的串扰所涉及的因子和分子机制，并强调了IFN刺激基因15（ISG15）的作用，它似乎处于这一交叉的中心。此外，我们还讨论了病原体感染时以及癌症和自身炎症综合征等人类疾病中免疫介导的 DNA 复制和修复调节的潜在意义和临床影响。最后，我们讨论了相关的未决问题和未来方向。

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引用次数: 0

Intersection of the fragile X-related disorders and the DNA damage response 脆性 X 相关疾病与 DNA 损伤反应的交集。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-07 DOI: 10.1016/j.dnarep.2024.103785

Daman Kumari , Jessalyn Grant-Bier , Farid Kadyrov , Karen Usdin

The Repeat Expansion Diseases (REDs) are a large group of human genetic disorders that result from an increase in the number of repeats in a disease-specific tandem repeat or microsatellite. Emerging evidence suggests that the repeats trigger an error-prone form of DNA repair that causes the expansion mutation by exploiting a limitation in normal mismatch repair. Furthermore, while much remains to be understood about how the mutation causes pathology in different diseases in this group, there is evidence to suggest that some of the downstream consequences of repeat expansion trigger the DNA damage response in ways that contribute to disease pathology. This review will discuss these subjects in the context of the Fragile X-related disorders (aka the FMR1 disorders) that provide a particularly interesting example of the intersection between the repeats and the DNA damage response that may also be relevant for many other diseases in this group.

重复扩增疾病（REDs）是一大类人类遗传疾病，是由于疾病特异性串联重复或微卫星的重复次数增加而导致的。新的证据表明，这些重复序列会引发一种容易出错的 DNA 修复形式，从而利用正常错配修复的局限性导致扩增突变。此外，尽管人们对突变如何导致该组不同疾病的病理变化仍有很多不解，但有证据表明，重复扩增的一些下游后果会触发 DNA 损伤反应，从而导致疾病的病理变化。本综述将以脆性 X 相关疾病（又称 FMR1 相关疾病）为背景讨论这些问题，该疾病提供了一个特别有趣的例子，说明了重复与 DNA 损伤反应之间的交叉关系，而这种交叉关系可能也与这类疾病中的许多其他疾病有关。

引用次数: 0

Discovery of KPT-6566 as STAG1/2 Inhibitor sensitizing PARP and NHEJ Inhibitors to suppress tumor cells growth in vitro 发现 KPT-6566 可作为 STAG1/2 抑制剂，使 PARP 和 NHEJ 抑制剂敏感，从而抑制肿瘤细胞在体外的生长。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-07 DOI: 10.1016/j.dnarep.2024.103784

Qinwei Zhu, Xuening Chen, Zhonghui Lin

Stromal antigen 1 and 2 (STAG1 and STAG2) are two mutually exclusive components of the cohesin complex that is crucial for centromeric and telomeric cohesion. Beyond its structural role, STAG2 also plays a pivotal role in homologous recombination (HR) repair and has emerged as a promising therapeutic target in cancer treatment. Here, we employed a fluorescence polarization (FP)-based high-throughput screening and identified KPT-6566 as a dual inhibitor of STAG1 and STAG2. Biochemical and biophysical analyses demonstrated that KPT-6566 directly binds to STAG1 and STAG2, disrupting their interactions with SCC1 and double-stranded DNA. A metaphase chromosome spread assay showed that KPT-6566 causes premature chromosome separation and induces chromosome damages in HeLa cells. Furthermore, KPT-6566 also impairs DNA damage repair, leading to the accumulation of double-strand breaks and cell apoptosis. Finally, KPT-6566 can sensitize HeLa and HepG2 cells to PARP inhibitor Olaparib and the NHEJ inhibitor UMI-77, exhibiting a synergistic effect in suppressing cell proliferation. Our findings highlight the potential of STAG1/2 as promising therapeutic targets in cancer treatment, particularly when they are targeted in combination with other DNA damage response inhibitors.

基质抗原 1 和 2（STAG1 和 STAG2）是凝聚素复合物的两个互斥成分，对中心粒和端粒的凝聚至关重要。除了其结构作用外，STAG2 还在同源重组（HR）修复中发挥着关键作用，并已成为癌症治疗中一个很有前景的治疗靶点。在这里，我们采用了基于荧光偏振（FP）的高通量筛选方法，发现 KPT-6566 是 STAG1 和 STAG2 的双重抑制剂。生化和生物物理分析表明，KPT-6566 能直接与 STAG1 和 STAG2 结合，破坏它们与 SCC1 和双链 DNA 的相互作用。转移期染色体扩散试验表明，KPT-6566 会导致 HeLa 细胞中染色体过早分离并诱发染色体损伤。此外，KPT-6566 还会损害 DNA 损伤修复，导致双链断裂积累和细胞凋亡。最后，KPT-6566 能使 HeLa 和 HepG2 细胞对 PARP 抑制剂 Olaparib 和 NHEJ 抑制剂 UMI-77 敏感，在抑制细胞增殖方面表现出协同效应。我们的研究结果凸显了 STAG1/2 作为癌症治疗靶点的潜力，尤其是当它们与其他 DNA 损伤反应抑制剂联合使用时。

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引用次数: 0

The biochemistry of the carcinogenic alcohol metabolite acetaldehyde 致癌酒精代谢物乙醛的生物化学。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-05 DOI: 10.1016/j.dnarep.2024.103782

Liam A. Thomas, Richard J. Hopkinson

Acetaldehyde (AcH) is the first metabolite of ethanol and is proposed to be responsible for the genotoxic effects of alcohol consumption. As an electrophilic aldehyde, AcH can form multiple adducts with DNA and other biomolecules, leading to function-altering and potentially toxic and carcinogenic effects. In this review, we describe sources of AcH in humans, including AcH biosynthesis mechanisms, and outline the structures, properties and functions of AcH-derived adducts with biomolecules. We also describe human AcH detoxification mechanisms and discuss ongoing challenges in the field.

乙醛（Acetaldehyde，ACH）是乙醇的第一种代谢产物，被认为是造成饮酒遗传毒性效应的原因。作为一种亲电醛，AcH 可与 DNA 和其他生物大分子形成多种加合物，导致功能改变，并可能产生毒性和致癌作用。在这篇综述中，我们描述了人类体内 AcH 的来源，包括 AcH 的生物合成机制，并概述了 AcH 与生物大分子生成的加合物的结构、性质和功能。我们还描述了人类的 AcH 解毒机制，并讨论了该领域目前面临的挑战。

引用次数: 0

One-ended and two-ended breaks at nickase-broken replication forks 缺口酶断裂复制叉上的单端断裂和双端断裂

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-04 DOI: 10.1016/j.dnarep.2024.103783

Ralph Scully , Johannes C. Walter , André Nussenzweig

Replisome collision with a nicked parental DNA template can lead to the formation of a replication-associated double strand break (DSB). How this break is repaired has implications for cancer initiation, cancer therapy and therapeutic gene editing. Recent work shows that collision of a replisome with a nicked DNA template can give rise to either a single-ended (se) or a double-ended (de)DSB, with potentially divergent effects on repair pathway choice and genomic instability. Emerging evidence suggests that the biochemical environment of the broken mammalian replication fork may be specialized in such a way as to skew repair in favor of homologous recombination at the expense of non-homologous end joining.

复制体与有缺口的亲代 DNA 模板碰撞会形成与复制相关的双链断裂（DSB）。如何修复这种断裂对癌症的诱发、癌症治疗和治疗性基因编辑都有影响。最近的研究表明，复制体与切口 DNA 模板的碰撞可产生单端（se）或双端（de）DSB，对修复途径选择和基因组不稳定性可能产生不同的影响。新出现的证据表明，哺乳动物复制叉断裂的生化环境可能具有特异性，以牺牲非同源末端连接为代价，偏向于同源重组修复。

引用次数: 0

Transient HR enhancement by RAD51-stimulatory compound confers protection on intestinal rather than hematopoietic tissue against irradiation in mice RAD51 激化化合物对 HR 的短暂增强可保护小鼠肠道组织而非造血组织免受辐照。

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair

Pub Date : 2024-11-02 DOI: 10.1016/j.dnarep.2024.103781

Zhiyu Lu , Dong Chen , Ning Zhang , Zhiyuan Zheng , Zimo Zhou , Guochen Liu , Jiawei An , Yong Wang , Yongping Su , Wensheng Chen , Fengchao Wang

DNA double-strand breaks (DSBs) are cytotoxic lesions that compromise genomic integrity and trigger cell death. Homologous recombination (HR) is a major pathway for repairing DSBs in cycling cells. However, it remains unclear whether transient modulation of HR could confer protection to adult stem cells against lethal irradiation exposure. In this study, we investigated the radio-protective effect of the RAD51-stimulatory compound RS-1 on adult stem cells and progenitor cells with varying cycling rates in intestinal and hematopoietic tissues. Treatment with RS-1 even at high doses did not induce noticeable cell death or proliferation of intestinal crypt cells in vivo. Pretreatment with RS-1 before irradiation significantly decreased mitotic death, promoted DNA repair and enhanced the survival of intestinal stem cells and progenitor cells and increased the number of regenerative crypt colonies thereby mitigating IR-induced gastrointestinal syndrome. Moreover, RS-1 pretreatment could increase the survival and regeneration of irradiated intestinal organoids in vitro, which can be rescued by RAD51 inhibitor. However, pretreatment with RS-1 in vivo did not elevate nucleated cell count or HSPCs in bone marrow after 6 Gy irradiation. Additionally, there was no impact on mouse survival due to drug treatment observed. Thus, our data suggest that targeting HR as a strategy to prevent tissue damage from acute irradiation exposure may depend on cell cycling rates and intrinsic DNA repair mechanisms.

DNA双链断裂（DSB）是一种细胞毒性损伤，会损害基因组的完整性并引发细胞死亡。同源重组（HR）是修复循环细胞中DSB的主要途径。然而，瞬时调节HR是否能保护成体干细胞免受致命辐照照射，目前仍不清楚。在这项研究中，我们研究了RAD51刺激化合物RS-1对肠道和造血组织中不同循环速率的成体干细胞和祖细胞的放射保护作用。即使使用高剂量的RS-1，在体内也不会引起肠隐窝细胞明显的细胞死亡或增殖。辐照前用RS-1预处理可显著减少有丝分裂死亡，促进DNA修复，提高肠道干细胞和祖细胞的存活率，增加隐窝再生集落的数量，从而减轻红外诱导的胃肠道综合征。此外，RS-1预处理可提高体外辐照肠组织细胞的存活率和再生率，RAD51抑制剂可挽救这种情况。然而，在体内用RS-1预处理并不能提高6 Gy辐照后骨髓中的有核细胞数或HSPCs。此外，药物治疗也没有影响小鼠的存活率。因此，我们的数据表明，以HR为靶点作为防止急性辐照造成组织损伤的策略可能取决于细胞循环率和内在DNA修复机制。

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