Cross-talking pre-incision events of eukaryotic NER

真核 NER 的串扰切前事件

基本信息

批准号：
7436230
负责人：
ALTAF A WANI
金额：
$ 32.99万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-06-01 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7436230
关键词：
Acetylation Address Arts Biochemical Biochemical Pathway Cataloging Catalogs Cell Line Cell Nucleus Cells Chromatin Co-Immunoprecipitations Complex Condition Crowding DDB Chlorinated Hydrocarbons DNA DNA Damage DNA Repair DNA Sequence DNA lesion DNA-Binding Proteins Deltastab Dependence EP300 gene Enabling Factors Environmental Exposure Eukaryota Eukaryotic Cell Event Excision Excision Repair Gene Targeting Genome Genome Stability Health Histone Acetylation Histone H3 Histones Human Immunoprecipitation In Situ In Vitro Laboratories Lesion Link Maintenance Mediating Molecular Nature Nuclear Nucleotide Excision Repair Pathway interactions Phase Placement Platelet Factor 4 Polymerase Chain Reaction Process Proteins Range Recording of previous events Recruitment Activity Regulatory Pathway Role ST13 gene Site Spottings Surgical incisions System TP53 gene Technology Transcription-Coupled Repair Transferase Work XPA gene Xenobiotics aplidine base chromatin remodeling design grasp in vivo insight intracellular protein transport mammalian genome protein localization location repaired research study transcription factor ultraviolet irradiation ultraviolet lesions

项目摘要

DESCRIPTION (provided by applicant): The maintenance of genomic stability in eukaryotes is relentlessly challenged by DNA damage. In order to understand the basis of damage tolerance and avoidance of deleterious effects, it is important to grasp not only the nature of such DNA damage and its processing pathways in cells, but also to have a clear understanding of how cells manage the lesion repair in the highly compact and tightly regulated chromatin milieu of the eukaryotic nucleus. This proposal is based on the premise that the specific biochemical pathways, relying on wide-ranging inter-molecular cross-talk between the pathway components, operate at a higher efficiency under conditions of macromolecular crowding and sequentially recruit specific factors for access a s well as repair of genotoxic lesions within mammalian genome. The proposal builds on our recent observations that support a strong link between DNA repair and p53 mediated and other related regulatory pathways. The overall experiments are designed to address the specific hypothesis that cellular DNA damage recognition and transcription factors, congregate at DNA lesions sites, open the chromatin via histone acetylation and initiate the key pre-incision events of nucleotide excision repair (NER). The proposed work will utilize the PI's established expertise in relevant, cellular, molecular, biochemical, and immunological technologies to undertake the following specific aims. (1) To assess the nature and extent of contribution of damaged DNA binding protein, DDB, in NER through quantitative assessment of NER sub-pathways in human cells of defined protein status. (2) To demonstrate the role of DDB for sequential recruitment of initial recognition and downstream core NER factors through spatial and temporal co-localization in situ. (3) To determine the participation of histone acetyl transferases in damage recognition and excision via qualitative protein cataloging and targeted recruitment and physical association of interacting factors. (4) To establish the histones acetylation as a direct and key consequence of DNA damage by placement of acetylation events exclusively at the lesion sites in vivo and in vitro. These studies will provide important insights regarding the dynamics of DNA damage recognition and processing through identification of cross-talking events and critical factors that influence the maintenance of genomic stability so important to overcome the hazardous health effects of genotoxic environmental exposures.

描述（由申请人提供）：在真核生物中的基因组稳定性维持受到DNA损伤的不懈挑战。为了理解损害耐受性和避免有害影响的基础，不仅要掌握这种DNA损伤的性质及其在细胞中的处理途径，而且要清楚地了解细胞在高度紧凑和紧密调控的真实核神经核的染色质环境中如何管理病变修复。该提案基于这样的前提：特定的生化途径依赖于途径成分之间的广泛分子间交叉对话，在较高的效率下，在雄性分子拥挤的条件下以较高的效率运行，并依次募集特定因素来访问哺乳动物基因组中遗传毒性病变的S e and s and s s and s s s s s s n s replate哺乳动物基因组中的修复。该提案建立在我们最近的观察结果上，该观察支持DNA修复与p53介导的和其他相关调节途径之间的牢固联系。整体实验旨在解决以下特定假设：细胞DNA损伤识别和转录因子聚集在DNA病变位点，通过组蛋白乙酰化打开染色质，并启动核苷酸切除修复的关键前事件（NER）。拟议的工作将利用PI在相关，细胞，分子，生化和免疫技术方面的既定专业知识来实现以下特定目标。（1）通过定量评估定义蛋白质状态的人类细胞中NER子街道的定量评估，在NER中评估受损DNA结合蛋白DDB的贡献的性质和程度。（2）通过空间和时间共定位的原位来证明DDB在序列募集初始识别和下游核心因素方面的作用。（3）确定组蛋白乙酰基转移酶通过定性蛋白质分解以及相互作用因子的靶向募集以及物理关联而参与损害识别和切除。（4）建立组蛋白乙酰化作为DNA损伤的直接和关键结果，这是通过在体内和体外的病变部位放置乙酰化事件的直接和关键结果。这些研究将通过识别跨言语事件和影响维持基因组稳定性的关键因素来提供有关DNA损伤识别和处理动态的重要见解，这些事件和关键因素对克服基因毒性环境暴露的危险健康效应至关重要。