The role of And-1 in nucleotide excision repair

And-1在核苷酸切除修复中的作用

• 批准号: 10576346
• 负责人: Wenge Zhu
• 金额: $ 44.67万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-17 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10576346
• 关键词: Acceleration; Affect; Animal Model; Binding; Cell Cycle; Cells; Chromosomal Stability; Complement; DBL Oncoprotein; DNA Damage; DNA Repair; DNA biosynthesis; DNA lesion; DNA-Binding Proteins; DNA-Directed DNA Polymerase; Data; Dermal; Excision Repair; Fibroblasts; Future; Genes; Genetic Transcription; Genome; Genome Stability; Knowledge; Lesion; Ligation; Link; Malignant Neoplasms; Mammals; Mediating; Molecular; Mus; Mutate; Mutation; Nuclear; Nucleoplasm; Nucleotide Excision Repair; Nucleotides; Orthologous Gene; Pathway interactions; Patients; Phosphorylation; Physiological; Play; Polymerase; Positioning Attribute; Precision therapeutics; Process; Protein Conformation; Proteins; Regulation; Regulatory Pathway; Role; Series; Single-Stranded DNA; Site; Skin; Skin Cancer; Skin Neoplasms; Testing; Time; Transcription-Coupled Repair; UV induced; UV induced DNA damage; Ultraviolet Rays; Xeroderma Pigmentosum; Yeasts; chemotherapeutic agent; design; environmental mutagens; experimental study; gene repair; homologous recombination; in vivo; individualized prevention; mouse model; multidisciplinary; novel; recombinational repair; recruit; repaired; response; skin squamous cell carcinoma; therapeutic development; tumorigenesis; ultraviolet irradiation; ultraviolet lesions

Project Summary Nucleotide excision repair (NER) is the major pathway to remove bulky DNA lesions induced by UV irradiation, environmental mutagens, and chemotherapeutic agents. Deficiency of genes involved in NER has been linked to Xeroderma Pigmentosum (XP) and skin cancer. There are two mechanisms to detect DNA damage by NER, one is global genome NER(GG-NER) and another is transcription-coupled NER(TC-NER). GG-NER occurs anywhere in the genome, whereas TC-NER is responsible for the accelerated repair of lesions in the transcribed strand of active genes. The both pathways are divided into early and late steps. The early step is the sequential actions, in which XP proteins recognize, unwind, and incise the DNA lesion. The latter step is identical in both mechanisms and is characterized by gap-filling repair synthesis, in which DNA replication proteins fill in the ~30 nucleotide gap, followed by ligation. Compared to the well-characterized early step, the molecular mechanism regulating the activation of gap-filling DNA synthesis at late step remains largely unknown. Even less is known about the physiological impact of such a regulatory pathway. And-1 is an acidic nucleoplasmic DNA-binding protein and its yeast ortholog, Ctf4, was originally identified as a critical gene for chromosome stability. Interestingly, yeast cells with depletion of Ctf4 gene are hypersensitive to UV lights, suggesting a role of And-1 in UV-induced DNA damage response. However, how And-1 regulates NER remains largely unknown. In this study, we now have extensive preliminary data demonstrating that And-1 is critical for NER by regulating gap-filling DNA synthesis. Our hypothesis is that And-1 regulates DNA polymerase activity at UV-lesion sites to activate gap-filling DNA synthesis at the late stage of NER. To test this hypothesis, we plan to pursue three specific aims. Aim 1: Determine the mechanism by which And-1 regulates DNA polymerase activity in NER. Aim 2: Determine the unique mechanism by which And-1 is recruited to UV-lesion sites. Aim 3: Determine the role of And-1 in NER and skin tumorigenesis using And-1 deficient mouse models. The completion of proposed studies will not only advance the field by uncovering a novel And-1-mediated pathway to regulate NER, but also provide us with the in vivo evidence to elucidate a novel role of And-1 in NER and skin tumor.

项目摘要 核苷酸切除修复（NER）是去除紫外线照射引起的DNA损伤的主要途径， 环境诱变剂和化疗剂。NER相关基因的缺失与 着色性干皮病（XP）和皮肤癌。NER检测DNA损伤有两种机制， 一种是全局基因组NER（GG-NER），另一种是转录偶联NER（TC-NER）。GG-NER发生 在基因组中的任何地方，而TC-NER负责加速修复基因组中的损伤。 活性基因的转录链。这两种途径分为早期和晚期步骤。第一步是 XP蛋白识别、解开和切割DNA损伤的顺序动作。后一步是 在两种机制中是相同的，其特征在于间隙填充修复合成，其中DNA复制 蛋白质填充约30个核苷酸的缺口，然后连接。与特征明确的早期步骤相比， 在后期调节间隙填充DNA合成激活的分子机制仍然很大程度上 未知对这种调节途径的生理影响所知更少。 And-1是一种酸性核质DNA结合蛋白，其酵母直向同源物Ctf 4最初是 被鉴定为染色体稳定性的关键基因。有趣的是，具有Ctf 4基因缺失的酵母细胞在细胞中表达。 对UV光过敏，提示And-1在UV诱导的DNA损伤反应中的作用。但如何 And-1调节NER仍然是未知的。在这项研究中，我们现在有大量的初步数据， 这表明And-1通过调节间隙填充DNA合成对NER至关重要。我们的假设是 And-1调节紫外线损伤部位的DNA聚合酶活性，以激活晚期的间隙填充DNA合成。 阶段。为了验证这一假设，我们计划追求三个具体目标。目标1：确定机制 And-1通过其调节NER中DNA聚合酶的活性。目标2：确定 和-1被募集到UV损伤部位。目的3：使用免疫组织化学方法确定And-1在NER和皮肤肿瘤发生中的作用。 和-1缺陷小鼠模型。完成拟议的研究不仅将推动该领域的发展， 揭示了一种新的And-1介导的调节NER的途径，但也为我们提供了体内证据， 阐明An-1在NER和皮肤肿瘤中的新作用。