Biochemical Insight of Hyperphosphorylation of Replication Protein A

复制蛋白 A 过度磷酸化的生化见解

• 批准号: 7365510
• 负责人: Yue Zou
• 金额: $ 20.89万
• 依托单位: EAST TENNESSEE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7365510
• 关键词: A-Form DNA; Academic Research Enhancement Awards; Address; Affect; Affinity; Appendix; Binding; Biochemical; Biochemical Pathway; Biological; Biomedical Research; Cancer Biology; Cell Cycle Progression; Cell physiology; Cells; Characteristics; Chemicals; DNA; DNA Binding Domain; DNA Damage; DNA Repair; DNA biosynthesis; DNA damage checkpoint; DNA lesion; Data; Defect; Disease; Environment; Environmental Hazards; Fire - disasters; Fluorescence Resonance Energy Transfer; Fluorescence Spectroscopy; Genetic; Genetic Recombination; Genomic Instability; Goals; Human; Laboratories; Lead; Malignant Neoplasms; Mediating; Metabolic Pathway; Metabolism; Methods; Molecular; Mutagenesis; Mutation; Nerve Degeneration; Phosphorylation; Play; Prevention; Process; Property; Proteins; Regulation; Replication Initiation; Research; Role; SS DNA BP; Series; Signal Transduction; Site; Site-Directed Mutagenesis; Source; Stress; Students; Surface Plasmon Resonance; Testing; Thermodynamics; Time; Work; base; carcinogenesis; career; coping; experience; insight; medical schools; repaired; replication factor A; response; ultraviolet irradiation

DESCRIPTION (provided by applicant): Summary The goal of this project is to elucidate the biochemical basis of the functions of human replication protein A (RPA) in DNA damage responses, and to better understand the damage-induced mutagenesis and carcinogenesis. DNA damage has been widely implicated in human cancer induction. Human RPA, the major single-stranded DNA binding protein, is essential for all DNA metabolic pathways such as DNA replication, DNA repair, recombination and DNA damage checkpoints. Given that RPA undergoes hyperphosphorylation in cells in response to DNA damage, it is hypothesized that the protein may play a modulatory role in cellular DNA damage responses mediated by the hyperphosphorylation at the N- terminus of its RPA32 subunit. How the hyperphosphorylation affect the cellular activities of RPA in terms of biochemical mechanism remains unknown. In this study, using an array of rigorous biochemical methods we will: (1) identify domain-domain interactions and structural characteristic of RPA upon DNA damage-induced hyperphosphorylation; (2) characterize the effects of hyperphosphorylation on RPA interaction with DNA intermediates of biological significance; and (3) determine binding affinity and thermodynamics of the hyperphosphorylation-induced domain-domain interaction of RPA. The proposed study is highly relevant to cancer biology as human defects in DNA damage responses lead to genome instability and thus a series of diseases with high potential to evolve cancer. This Academic Research Enhancement Award will allow undergraduate and graduate students to gain biomedical research experience in a medical school environment that will prepare them for careers in scientific research. Specifically, the proposed study will address an important question regarding the biochemical basis of RPA hyperphosphorylation in cellular DNA damage responses. This project is highly relevant to cancer biology as DNA damage is the major cause to human cancer.

描述（由申请人提供）：总结该项目的目的是阐明人类复制蛋白A（RPA）在DNA损伤反应中的功能的生化基础，并更好地了解损伤诱导的诱变诱变和癌变。 DNA损伤已广泛与人类癌症诱导有关。人RPA是主要的单链DNA结合蛋白，对于所有DNA代谢途径，例如DNA复制，DNA修复，重组和DNA损伤检查点都是必不可少的。鉴于RPA响应DNA损伤会在细胞中经历过度磷酸化，因此假设该蛋白可能在其RPA32亚基的N-末端介导的细胞DNA损伤反应中起调节作用。高磷酸化如何影响RPA的生化机制的细胞活性仍然未知。在这项研究中，使用一系列严格的生化方法，我们将：（1）在DNA损伤诱导的过度磷酸化时识别RPA的结构域相互作用和结构特征； （2）表征了高磷酸化对RPA相互作用与生物学意义的DNA中间体的影响； （3）确定过度磷酸化诱导的RPA结构域相互作用的结合亲和力和热力学。拟议的研究与癌症生物学高度相关，因为DNA损伤反应中的人缺陷导致基因组不稳定性，因此一系列具有高潜力进化癌症的疾病。这项学术研究增强奖将使本科生和研究生能够在医学院环境中获得生物医学研究经验，这将为科学研究的职业做好准备。具体而言，拟议的研究将解决有关细胞DNA损伤反应中RPA高磷酸化的生化基础的一个重要问题。该项目与癌症生物学高度相关，因为DNA损伤是人类癌症的主要原因。