Mechanism of APE2 in genome integrity

APE2在基因组完整性中的机制

• 批准号: 10578464
• 负责人: Shan Yan
• 金额: $ 7.43万
• 依托单位: UNIVERSITY OF NORTH CAROLINA CHARLOTTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10578464
• 关键词: APEXL2 Gene; Aging; Biochemical; Bioinformatics; Cancer Etiology; Cells; Cellular biology; Chemotherapy-Oncologic Procedure; Complex; DNA; DNA Damage; DNA Repair; DNA Sequence Alteration; DNA Structure; DNA glycosylase; DNA metabolism; Data; Deoxyribose; Double-Stranded RNA; Excision; Exonuclease; Feedback; Frequencies; Genetic Transcription; Genome Stability; Hybrids; Hydroxyl Radical; Lesion; Malignant neoplasm of liver; Malignant neoplasm of lung; Mediating; Metabolism; Methyltransferase; Modification; Molecular; Molecular Biology; Mutagenesis; NEIL3 gene; Neurodegenerative Disorders; Non-Malignant; Nucleic Acids; Oncogenes; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Phosphodiesterase I; Plasmids; Play; Process; Proteins; Publishing; RNA; RNA Processing; RNA metabolism; Reactive Oxygen Species; Renal carcinoma; Reporting; Research; Role; Saccharomycetales; Series; Single Strand Break Repair; Single-Stranded DNA; Site; Structure; System; Testing; Tissues; Toxic Environmental Substances; Transcriptional Regulation; Translations; Tumor Tissue; Uterine Cancer; Xenopus; base; brca gene; cancer cell; cancer therapy; cancer type; egg; genome integrity; inorganic phosphate; insight; malignant breast neoplasm; novel; nucleic acid metabolism; oxidative DNA damage; preservation; protein metabolism; protein purification; reconstitution; repaired; response; tumorigenesis

Project Summary/Abstract Environmental toxins or endogenous insults such as reactive oxygen species (ROS) result in oxidative stress within cells, leading to different types of damage in DNA or RNA, including Apurinic/apyrimidinic (AP) sites and single-strand breaks (SSBs). As the most common type of DNA damage, SSBs have been implicated in association with tumorigenesis, aging, and neurodegenerative disorders. Molecular understanding of SSB repair pathway remains unclear, largely due to the lack of tractable experimental systems. We and others have demonstrated recently that APE2 resolves SSB damage in Xenopus and budding yeast. Although APE1 has been found critical for including DNA repair, redox regulation of transcription, and RNA processing, it remains unclear how APE2 plays important roles in DNA and RNA metabolism. It is significant to determine how APE2 plays an essential role in SSB repair, and how exactly APE2 maintains genome stability. Our substantial preliminary data suggest that APE2’s 3'-5' exonuclease activity is regulated via its dynamic interactions with single-strand DNA (ssDNA) and its interacting proteins, and that APE2 associates with RNA and RNA-containing structures such as R-loop. We will dissect the molecular mechanism of APE2 in metabolism in nucleic acids via two Specific Aims: (1) determine the mechanism of how APE2 interacts with and processes AP sites and SSBs in DNA structures in reconstitution system with purified proteins and in Xenopus egg extracts, and (2) determine how APE2 recognizes and repairs AP site and SSBs in RNA or RNA-containing structures. Biochemical, cell biology, and molecular biology approaches will be utilized to conduct this hypothesis-driven structure-function analysis of APE2-mediated metabolism of nucleic acids in Xenopus egg extracts and reconstitution systems with purified proteins. Anticipated results from this project will make a paradigm shift on APE2- mediated SSB repair in DNA/RNA metabolism to maintain genome stability. Notably, our findings will provide novel insights into new strategies for cancer chemotherapies such as modulating the distinct regulatory mechanisms of APE2 for SSB repair in genome integrity.

项目概要/摘要 环境毒素或内源性损伤（例如活性氧 (ROS)）会导致氧化 细胞内的压力，导致 DNA 或 RNA 的不同类型的损伤，包括无嘌呤/无嘧啶 (AP) 位点和单链断裂 (SSB) 作为最常见的 DNA 损伤类型，SSB 已成为最常见的 DNA 损伤类型。 与肿瘤发生、衰老和分子退行性疾病有关。 对 SSB 修复途径的理解仍不清楚，很大程度上是由于缺乏易于处理的实验 我们和其他人最近已经证明 APE2 可以解决非洲爪蟾和非洲爪蟾的 SSB 损伤。 尽管APE1被发现对于DNA修复、氧化还原调节至关重要。 转录和 RNA 加工，目前尚不清楚 APE2 如何在 DNA 和 RNA 中发挥重要作用 确定 APE2 在 SSB 修复中如何发挥重要作用以及到底如何发挥重要作用。 APE2 保持基因组稳定性。我们的大量初步数据表明 APE2 的 3'-5' 核酸外切酶。 活性通过其与单链 DNA (ssDNA) 及其蛋白质相互作用的动态相互作用进行调节， APE2 与 RNA 和包含 RNA 的结构（例如 R 环）相关联，我们将对其进行剖析。 通过两个具体目标研究APE2在核酸代谢中的分子机制：（1）确定 APE2如何与DNA结构中的AP位点和SSB相互作用并对其进行处理的机制 用纯化的蛋白质和爪蟾卵提取物重建系统，以及 (2) 确定 APE2 如何 识别并修复 RNA 或含 RNA 结构中的 AP 位点和 SSB。 分子生物学方法将用于进行这种假设驱动的结构功能 爪蟾卵提取物和重构中 APE2 介导的核酸代谢分析 该项目的预期结果将使 APE2- 发生范式转变。 介导 DNA/RNA 代谢中的 SSB 修复以维持基因组稳定性。 为癌症化疗的新策略提供新颖的见解，例如调节不同的 APE2 对基因组完整性 SSB 修复的调节机制。