Mechanisms of DNA damage processing and the initiation of Nucleotide Excision Repair

DNA损伤处理机制和核苷酸切除修复的启动

基本信息

批准号：
10513526
负责人：
Jung-Hyun Min
金额：
$ 37.75万
依托单位：
BAYLOR UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10513526
关键词：
2-Acetylaminofluorene 3-Dimensional Antineoplastic Agents Binding Biochemical Biological Biomedical Research Biophysics Carcinogens Cell physiology Cells Chemotherapy-Oncologic Procedure Chromatin Circular DNA Cisplatin Clinical Cockayne Syndrome Collaborations Complex Cryoelectron Microscopy DNA DNA Adducts DNA Damage DNA Repair DNA Repair Pathway DNA lesion DNA-Protein Interaction Defect Disease ERCC3 gene Electron Microscope Environment Excision Repair Fluorescence Fluorescence Resonance Energy Transfer Fluorescence Spectroscopy Foundations Functional disorder Genetic Transcription Genomic DNA Genomic Instability Goals Homologous Gene Human In Vitro Institution Interdisciplinary Study Lead Lesion Licensing Link Malignant Neoplasms Maps Mass Spectrum Analysis Measurement Medical Methodology Modeling Molecular Molecular Conformation Multiprotein Complexes Mutation Nucleotide Excision Repair Outcome Outcomes Research Pathologic Pathway interactions Pennsylvania Phenotype Premature aging syndrome Process Proteins RAD23B gene Research Research Personnel Research Project Grants Research Support Research Training Resistance Resolution Science Site Solid Somatic Mutation Stress Structure Sunlight Syndrome Technology Time Torsion Training Trichothiodystrophy Ultraviolet Rays Universities Xeroderma Pigmentosum Yeasts anti-cancer therapeutic base biophysical techniques cancer predisposition cancer therapy computerized tools crosslink environmental mutagens genome integrity global genomic repair medical schools novel novel strategies novel therapeutic intervention reconstitution recruit repaired sensor structural biology success synergism three dimensional structure transcription factor S-II translocase transmission process ultraviolet lesions undergraduate student

项目摘要

Project Summary: Mechanisms of DNA damage processing and the initiation of Nucleotide Excision Repair The goal of this research is to determine the structural mechanism of nucleotide excision repair (NER) initiation. NER is the most versatile DNA repair mechanism that repairs a wide variety of DNA lesions through a multistep process involving over 30 different proteins. Being essential to maintaining genome integrity, this pathway is also highly conserved from yeast to humans. Genetic defects in NER factors lead to phenotypes ranging from extreme cancer predisposition syndrome (xeroderma pigmentosum) to severe neurodevelopmental defects (Cockayne syndrome), thus providing a unique paradigm to understand diverse clinical outcomes of DNA damage. Recent studies also revealed NER as a major contributor of somatic mutation hotspots in various sporadic cancers and NER has been suggested as an attractive target for anti-cancer therapy. Despite its biological and medical importance, delineating the mechanisms of NER has been a long-term challenge due to the complex compositions and functions of NER factors and the lack of comprehensive structural understanding of their interplay on DNA. This proposal aims to define the mechanism of NER initiation in detailed 3D structures using cryo-EM combined with time- resolved fluorescence spectroscopy and crosslinking/mass-spectrometry. The outcome will answer fundamental questions regarding (1) how the two key NER initiators, Rad4-Rad23-Rad33 (yeast homolog of XPC-RAD23B-CETN2) and TFIIH, together start the DNA ‘opening’ around the damage - a critical step in NER initiation, and (2) how the torsional stress in DNA impacts this process. This understanding will provide the foundation to explain various pathophysiologies involving NER, which in turn can lead to novel strategies to counter various NER-linked diseases including cancer. Importantly, our research will provide solid training grounds for several undergraduate researchers every year and will significantly enhance the biomedical research environment at Baylor University, an undergraduate-focused institution, through its intimate collaboration with UPenn Medical School. Immersed in an interdisciplinary research project with access to cutting- edge technologies, our undergraduate researchers will gain expertise in various biochemical and biophysical approaches and grow as key drivers of significant science.

项目概要： DNA损伤处理机制和核苷酸切除修复的启动本研究的目标是确定核苷酸切除修复的结构机制 (NER) 启动是最通用的 DNA 修复机制，可修复多种DNA。 DNA 损伤是通过涉及 30 多种不同蛋白质的多步骤过程实现的。为了保持基因组完整性，该途径从酵母到人类也高度保守。 NER 因子的遗传缺陷导致从极端癌症易感性到各种表型综合征（着色性干皮病）至严重神经发育缺陷（Cockayne 综合征），从而提供了一个独特的范例来理解 DNA 的不同临床结果最近的研究还表明 NER 是体细胞突变热点的主要贡献者。在各种散发性癌症中，NER 被认为是一个有吸引力的抗癌靶点治疗。尽管 NER 具有生物学和医学重要性，但描述 NER 的机制一直是一个难题。由于 NER 因子的复杂组成和功能以及缺乏该提案旨在对它们在 DNA 上的相互作用进行全面的结构理解。使用冷冻电镜结合时间-在详细的 3D 结构中启动 NER 的机制解析荧光光谱和交联/质谱的结果将。回答有关 (1) 两个关键 NER 启动子 Rad4-Rad23-Rad33 如何进行的基本问题（XPC-RAD23B-CETN2 的酵母同源物）和 TFIIH，一起启动围绕损伤 - NER 启动的关键步骤，以及 (2) DNA 中的扭转应力如何影响这一步骤这种理解将为解释各种病理生理学提供基础。涉及 NER，这反过来又可以带来对抗各种 NER 相关疾病的新策略包括癌症。重要的是，我们的研究将为一些本科生提供坚实的训练基础每年都有研究人员加入，并将显着改善生物医学研究环境贝勒大学是一所以本科生为重点的机构，通过与宾夕法尼亚大学医学院沉浸在一个跨学科研究项目中，可以接触到前沿技术。边缘技术，我们的本科研究人员将获得各种生物化学和生物物理方法并成为重要科学的关键驱动力。