Repair Mechanisms For Oxidative DNA Damage

DNA 氧化损伤的修复机制

• 批准号: 9147336
• 负责人: David Wilson
• 金额: $ 39.35万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9147336
• 关键词: Aging; Aging-Related Process; Alkylation; Back; Base Excision Repairs; Binding; Biochemical; Cell Death; Cell Line; Cell Respiration; Cells; Chromosomal Stability; Cockayne Syndrome; Complement component C1; Complex; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA biosynthesis; DNA glycosylase; Disease; Disease susceptibility; Evolution; Excision; Exhibits; Family member; Flap Endonucleases; Foundations; Functional disorder; Genetic Transcription; Genome; Human; Immunoglobulin Class Switching; Immunoglobulin Switch Recombination; Individual; Lead; Lesion; Ligase; Link; Lipids; Location; Lyase; Maintenance; Mitochondria; Modification; Molecular; Molecular Conformation; Mutate; NPM1 gene; Nature; Nuclear; Nucleic Acids; Nucleosomes; Nucleotides; Organism; Oxidation-Reduction; Pathway interactions; Play; Polymerase; Population; Positioning Attribute; Post-Translational Protein Processing; Premature aging syndrome; Process; Proteins; RECQL4 gene; RECQL5 gene; RNA; Reactive Oxygen Species; Relative (related person); Repair Complex; Reporting; Research; Role; Rothmund-Thomson syndrome; Scaffolding Protein; Series; Single Strand Break Repair; Site; Surface; Surgical incisions; System; Toxic Environmental Substances; Uracil; Variant; Vertebral column; Work; age related; base; cell growth; chromatin remodeling; coping; crosslink; cytotoxic; disorder risk; endonuclease; genetically modified cells; helicase; inorganic phosphate; novel; nuclease; nucleophosmin; oxidative DNA damage; phosphodiester; repair enzyme; repaired; response; seal; tumor; uracil-DNA glycosylase

Using molecular, biochemical and structural approaches, we have broadly contributed to defining how specific human BER proteins recognize and process target lesions, as well as coordinate with other components of the protective system. The research has centered largely on apurinic/apyrimidinic endonuclease 1 (APE1), the major mammalian protein for repairing abasic sites in DNA, and x-ray cross-complementing 1 (XRCC1), a non-enzymatic scaffold protein that facilitates the efficient execution of single-strand break (SSB) repair. Some of the key findings during the course of the project include: (i) in addition to abasic sites in conventional duplex DNA, APE1 has the ability to incise at AP sites in DNA conformations formed during DNA replication, transcription, and class switch recombination, and APE1 can endonucleolytically destroy damaged RNA; (ii) APE1 contributes to the repair of 3-modifications in DNA, such as mismatches, phosphate groups, phosphogycolates and tyrosyl residues; (iii) the DNA repair function of APE1 is regulated in part by post-translational modification, such as S-glutathionylation; (iv) inhibition of APE1 is a potential mechanism for the genotoxic and co-carcinogenic effects of lead, an important environmental toxin; (v) APE1 communicates with CSB, a protein defective in the premature aging disorder, Cockayne syndrome; (vi) XRCC1 directly associates with the replication/repair protein, PCNA, establishing a novel link between the DNA repair machinery and replication factories; (vii) XRCC1 coordinates disparate responses and multi-protein repair complexes that are dependent on the nature and context of the DNA damage; (viii) the different regions of XRCC1 play distinct roles in coordinating repair complex assembly, and the population variant Arg280His exhibits reduced stability at DNA damage foci, suggesting that it may represent a disease susceptibility factor; (ix) XRCC1 supports an emerging pathway for uracil repair, termed replication-associated BER, through a physical association with UNG2, the major nuclear uracil DNA glycosylase; (x) the interaction of XRCC1 with the DNA repair enzyme PNKP functions to retain XRCC1 at DNA damage sites and to promote repair of alkylation damage; (xi) the flap-endonuclease FEN1 plays a role in repairing mitochondrial oxidative DNA damage through a long-patch BER pathway; (xii) RECQL4, a human RecQ helicase mutated in approximately two-thirds of individuals with Rothmund-Thomson syndrome, regulates BER capacity both directly and indirectly; and (xiii) RECQL5, another RECQ helicase family member, modulates and/or directly participates in BER of endogenous DNA damage, thereby promoting chromosome stability in normal human cells. Our most recent work has demonstrated that (i) the DNA glycosylase NEIL1 recognizes specifically ICLs in DNA, and can obstruct the efficient removal of these lethal lesions; (ii) the multifunctional protein nucleophosmin (a.k.a., NPM1) is a modulator of BER capacity by controlling BER protein levels and regulating the nucleolar localization of several BER enzymes; (iii) the efficiency of APE1 binding and strand incision are influenced by the domain sequence, conformation and AP site location/relative positioning within telomeric and CAG/CTG repeat sequences; and (iv) specific APE1 variants (e.g., the tumor-associated R237C variant) are uniquely hypersensitive to nucleosome complexes in the vicinity of the AP site, suggesting the evolution of distinct surface residues that permit efficient processing of abasic sites within the context of protein-DNA complexes independent of classic chromatin remodeling mechanisms. Currently, a main focus is to establish genetically modified cell lines to dissect out the precise contribution of each proposed function of APE1 (i.e. its nuclease activity, redox regulatory role, etc.) in cell growth/viability, genome maintenance, and protection against DNA-damaging agents. Defining which of the many reported functions of APE1 are critical to normal cellular activity is a key step towards understanding the potential relationship of the protein to the aging process and disease risk.

使用分子，生化和结构方法，我们广泛地促进了特定的人类BER蛋白如何识别和过程靶损伤，并与保护性系统的其他成分进行协调。 这项研究主要集中在肾上腺素/阿哌丁丁核酸内切酶1（APE1），这是用于修复DNA中的无碱性位点的主要哺乳动物蛋白，而X射线互补的1（XRCC1），一种非酶骨架蛋白，可促进单型骨架的有效执行。 项目过程中的一些关键发现包括：（i）除了传统的双链DNA中的无asic位点外，APE1还可以在DNA复制，转录和类开关重组中形成的DNA构象中的APENICS，APE1可以在核酸中销毁破坏的RNA； （ii）APE1有助于修复DNA中的3个修饰，例如不匹配，磷酸基团，磷酸盐和酪酶残基； （iii）APE1的DNA修复功能部分通过翻译后修饰（例如S-甲基二硫代）来调节； （iv）抑制APE1是铅（一种重要的环境毒素）的遗传毒性和共毒性作用的潜在机制； （v）APE1与CSB进行通信，CSB是Cockayne综合征过早衰老障碍中有缺陷的蛋白质； （vi）XRCC1与复制/修复蛋白PCNA直接关联，建立了DNA修复机械与复制工厂之间的新联系； （vii）XRCC1协调依赖于DNA损伤的性质和背景的不同反应和多蛋白修复复合物； （viii）XRCC1的不同区域在协调修复复合体组件中起着不同的作用，而种群变体ARG280HIS表现出DNA损伤焦点的稳定性降低，这表明它可能代表疾病的易感因子； （IX）XRCC1通过与UNG2的物理结合（主要的核尿尿嘧啶DNA糖基化酶）的物理结合，支持尿嘧啶修复的新兴途径，称为复制相关的BER； （x）XRCC1与DNA修复酶PNKP功能的相互作用可在DNA损伤位点保留XRCC1并促进烷基化损伤的修复； （xi）皮瓣 - 内核酸酶Fen1在通过长块ber途径修复线粒体氧化DNA损伤方面起作用； （XII）RECQL4，一种在大约三分之二的Rothmund-Thomson综合征中突变的人RECQ解旋酶，可以直接和间接地调节BER容量。 （XIII）RECQL5是另一位RECQ解旋酶家族成员，调节和/或直接参与内源性DNA损伤的BER，从而促进了正常人细胞中的染色体稳定性。 我们最近的工作表明（i）DNA糖基酶neil1在DNA中识别出特异性ICL，并且可以阻止有效去除这些致命病变； （ii）多功能蛋白质核素（又称NPM1）是通过控制BER蛋白水平并调节几种BER酶的核仁定位，是BER容量的调节剂； （iii）APE1结合和链切口的效率受域序列，构象和AP位点位置/相对位置的影响； （iv）特定的APE1变体（例如，与肿瘤相关的R237C变体）对AP位点附近的核小体复合物具有独特的高度敏感性，这表明在蛋白质-DNA复合物中有效地处理了具有有效的ABASIC位点的不同表面残基的演变，允许在蛋白质-DNA复合物中有效地处理经典的Chratees Complus Chropsect opplace Chratems Rematemations机制。 当前，主要重点是建立遗传修饰的细胞系，以阐明APE1的每个提出功能（即其核酸酶活性，氧化还原调节作用等）在细胞生长/生存力，基因组维持和针对DNA损害剂的保护中的精确贡献。 定义APE1的许多报告的功能中的哪个对正常细胞活性至关重要，这是了解蛋白质与衰老过程和疾病风险的潜在关系的关键步骤。

项目成果

Characterization of the endoribonuclease active site of human apurinic/apyrimidinic endonuclease 1.

• DOI: 10.1016/j.jmb.2011.06.050
• 发表时间: 2011-09-02
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Kim WC;Berquist BR;Chohan M;Uy C;Wilson DM 3rd;Lee CH
• 通讯作者: Lee CH

Direct interaction between XRCC1 and UNG2 facilitates rapid repair of uracil in DNA by XRCC1 complexes.

• DOI: 10.1016/j.dnarep.2010.04.002
• 发表时间: 2010-07-01
• 期刊: DNA repair
• 影响因子: 3.8
• 作者: Akbari M;Solvang-Garten K;Hanssen-Bauer A;Lieske NV;Pettersen HS;Pettersen GK;Wilson DM 3rd;Krokan HE;Otterlei M
• 通讯作者: Otterlei M

Nucleophosmin modulates stability, activity, and nucleolar accumulation of base excision repair proteins.

核磷蛋白调节碱基切除修复蛋白的稳定性、活性和核仁积累。

• DOI: 10.1091/mbc.e13-12-0717
• 发表时间: 2014
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Poletto,Mattia;Lirussi,Lisa;Wilson3rd,DavidM;Tell,Gianluca
• 通讯作者: Tell,Gianluca

A novel link to base excision repair?

• DOI: 10.1016/j.tibs.2010.01.003
• 发表时间: 2010-05
• 期刊: TRENDS IN BIOCHEMICAL SCIENCES
• 影响因子: 13.8
• 作者: Wilson, David M., III;Seidman, Michael M.
• 通讯作者: Seidman, Michael M.

The region of XRCC1 which harbours the three most common nonsynonymous polymorphic variants, is essential for the scaffolding function of XRCC1.

• DOI: 10.1016/j.dnarep.2012.01.001
• 发表时间: 2012-04-01
• 期刊: DNA REPAIR
• 影响因子: 3.8
• 作者: Hanssen-Bauer, Audun;Solvang-Garten, Karin;Gilljam, Karin Margaretha;Torseth, Kathrin;Wilson, David M., III;Akbari, Mansour;Otterlei, Marit
• 通讯作者: Otterlei, Marit