The Role of And-1 in DNA Damage Response

And-1 在 DNA 损伤反应中的作用

• 批准号: 8848359
• 负责人: Wenge Zhu
• 金额: $ 32.89万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8848359
• 关键词: Affect; Antineoplastic Agents; Applications Grants; Binding; Biochemical; Biological Assay; Cell Cycle; Cells; Complex; DNA; DNA Damage; DNA Repair; DNA Synthesis Inhibition; DNA biosynthesis; DNA replication fork; Data; Development; Event; Exhibits; Genes; Genetic Transcription; Genome Stability; Human; In Vitro; Knowledge; Link; Maintenance; Mammalian Cell; Mass Spectrum Analysis; Mediating; Molecular; Oncogenes; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological Processes; Play; Positioning Attribute; Process; Publishing; Radio; Recruitment Activity; Regulation; Replication Origin; Resistance; Role; Series; Signal Transduction; Signal Transduction Pathway; Site; Solid; Stress; Structure; Technology; Testing; Ultraviolet Rays; Work; carcinogenesis; design; in vivo; innovation; novel; prevent; public health relevance; response; single molecule; targeted cancer therapy; tumorigenesis; ultraviolet irradiation

DESCRIPTION (provided by applicant): DNA damage response (DDR) is a complex signal transduction pathway, which is extremely important to maintain genomic stability and prevent carcinogenesis by counteracting the deleterious effects of DNA damage. In response to replication stress or UV irradiation, the primary event of DDR is to activate kinases ATR and Chk1, which in turn trigger multiple physiological processes, including DNA synthesis inhibition, DNA repair, and cell cycle delay, transcription, etc. ATR plays a key role in Chk1 activation and DNA synthesis inhibition in response to DNA damage. However, how ATR coordinates with various DDR components to activate Chk1 and how ATR targets replication apparatus to inhibit DNA synthesis remain largely unknown in mammalian cells. We have demonstrated that human And-1 acts as a key component of replisome at replication forks for DNA replication. We now have solid preliminary data indicating that And-1 is also involved in the regulation of checkpoint and DNA synthesis in response to DNA damage. Thus, elucidating the role of And-1 in DDR will fill in critical knowledge gaps of DDR signaling. Building upon our recently published work and our extensive new data, in this proposal we described a series of innovative, hypothesis-driven studies to elucidate how human And-1 regulates checkpoint activation and DNA synthesis in response to UV irradiation and replication stress. First, we will determine the mechanism of how And-1 is recruited to DNA damage sites using both biochemical and structural analyses. Second, we will determine how And-1 acts as a unique replisome component to regulate checkpoint activation by impacting interplay among multiple key DDR components including ATR, Rad17, Claspin, and Timeless-Tipin at stalled replication forks using molecular and structural analyses. Finally, we will determine mechanism by which And-1 governs DNA synthesis in response to DNA damage by carrying out an innovative approach that combines multiple cutting edge technologies including single-molecule analyses, iPOND, and mass spectrometry. Given that targeting the circuitry of DNA damage response has been a key focus for the development of anti-cancer drugs, proposed work will not only advance the field by identifying new components and acquiring in-depth mechanistic understanding of DNA damage response, but also provide us with new strategies for the development of highly specific anti-cancer therapies targeting And-1 or And-1-dependent processes.

描述(申请人提供)：DNA损伤反应(DDR)是一种复杂的信号转导途径，通过抵消DNA损伤的有害影响，对维持基因组稳定和预防癌症发生极其重要。在对复制胁迫或紫外线辐射的反应中，DDR的主要事件是激活ATR和Chk1，进而触发多种生理过程，包括DNA合成抑制、DNA修复、细胞周期延迟、转录等，ATR在Chk1激活和DNA合成抑制中发挥关键作用。然而，在哺乳动物细胞中，ATR如何与不同的DDR组分协调激活Chk1，以及ATR如何靶向复制装置以抑制DNA合成在很大程度上仍是未知的。我们已经证明了人类和-1在DNA复制的复制叉处作为复制体的关键组成部分。我们现在有确凿的初步数据表明，And-1也参与了检查点和DNA合成的调节，以应对DNA损伤。因此，阐明和-1在DDR中的作用将填补DDR信号转导的关键知识空白。在我们最近发表的工作和我们广泛的新数据的基础上，在这个提案中，我们描述了一系列创新的、假设驱动的研究，以阐明人类和-1如何调节检查点激活和DNA合成，以响应紫外线照射和复制压力。首先，我们将利用生化和结构分析来确定和-1如何被招募到DNA损伤部位的机制。其次，我们将使用分子和结构分析来确定And-1如何作为一个独特的复制体组件，通过影响包括ATR、Rad17、Claspin和Timless-Tipin在内的多个关键DDR组件之间的相互作用来调节检查点激活。最后，我们将通过开展一种创新的方法，结合包括单分子分析、iPOND和质谱学在内的多种尖端技术，确定和-1调控DNA合成以应对DNA损伤的机制。鉴于靶向DNA损伤反应通路一直是抗癌药物开发的一个关键焦点，所提出的工作不仅将通过识别新的成分和深入了解DNA损伤反应的机制来推动该领域的发展，而且还将为靶向AND-1或AND-1依赖过程的高度特异性抗癌治疗的发展提供新的策略。