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

• 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
• 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

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.

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