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: 10362967
• 负责人: Wenge Zhu
• 金额: $ 45.58万
• 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/10362967
• 关键词: 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; Genome; Genome Stability; Hypersensitivity; Knowledge; Lesion; Ligation; Link; Malignant Neoplasms; Mammals; Mediating; Molecular; Mus; Mutate; Mutation; Nuclear; 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）是去除紫外线照射诱导的大量DNA病变的主要途径， 环境诱变剂和化学治疗剂。 NER涉及的基因缺乏已连接 进行静脉皮色素（XP）和皮肤癌。有两种检测NER DNA损伤的机制， 一个是全局基因组NER（GG-NER），另一个是转录耦合NER（TC-NER）。 GG-ner发生 基因组中的任何地方 活性基因的转录链。这两种途径都分为早期和晚期。早期的一步是 XP蛋白识别，放松和切除DNA病变的顺序作用。后一个步骤是 两种机制相同，并以间隙填充修复合成为特征，其中DNA复制 蛋白质填补了约30个核苷酸间隙，然后结扎。与特征良好的早期步骤相比 调节间隙填充DNA合成在后期的激活的分子机制在很大程度上保持不变 未知。关于这种调节途径的生理影响，知之甚少。 和-1是一种酸性核质DNA结合蛋白及其酵母直系同源物CTF4，最初是 被确定为染色体稳定性的关键基因。有趣的是，CTF4基因耗尽的酵母细胞是 对紫外线的高度敏感，表明和1在紫外线诱导的DNA损伤反应中的作用。但是，如何 和-1调节NER仍然在很大程度上未知。在这项研究中，我们现在拥有广泛的初步数据 通过调节间隙填充DNA合成，证明AND-1对于NER至关重要。我们的假设是 和-1调节紫外线质量位点的DNA聚合酶活性，以激活晚期的间隙填充DNA合成 ner阶段。为了检验这一假设，我们计划追求三个具体目标。目标1：确定机制 通过它调节NER中的DNA聚合酶活性。目标2：确定独特的机制 和-1被招募到紫外线矿场。 AIM 3：使用AND-1在使用NER和皮肤肿瘤发生中的作用 和-1不足的鼠标模型。拟议研究的完成不仅将通过 发现一种新颖的和1介导的途径来调节NER，但也为我们提供了体内证据 阐明和1在NER和皮肤肿瘤中的新作用。