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）：摘要本项目的目标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，并更好地理解损伤诱导的突变和致癌作用。 DNA 损伤与人类癌症的诱发广泛相关。人类 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）至关重要。鉴于 RPA 在细胞中因 DNA 损伤而发生过度磷酸化，因此推测该蛋白可能在由其 RPA32 亚基 N 末端过度磷酸化介导的细胞 DNA 损伤反应中发挥调节作用。过度磷酸化如何在生化机制方面影响 RPA 的细胞活性仍不清楚。在这项研究中，我们将使用一系列严格的生化方法：（1）识别DNA损伤诱导的过度磷酸化后RPA的域间相互作用和结构特征； (2) 表征过度磷酸化对 RPA 与具有生物学意义的 DNA 中间体相互作用的影响； (3)确定RPA过度磷酸化诱导的域-域相互作用的结合亲和力和热力学。这项拟议的研究与癌症生物学高度相关，因为人类 DNA 损伤反应的缺陷会导致基因组不稳定，从而导致一系列极有可能发展为癌症的疾病。该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，为他们从事科学研究职业做好准备。具体来说，拟议的研究将解决有关细胞 DNA 损伤反应中 RPA 过度磷酸化的生化基础的重要问题。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。