DNA-Protein Cross-link Formation by Chimeric Bis-electrophiles

嵌合双亲电子试剂形成 DNA-蛋白质交联

• 批准号: 2346706
• 负责人: Marc Greenberg
• 金额: $ 55.5万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2346706&HistoricalAwards=false
• 关键词: DNA; Protein; Cross; link; Formation; DNA; Protein; Cross; link; Formation

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry, Professor Marc Greenberg of Johns Hopkins University and his research team are studying the development of molecules for the selective formation of DNA-protein crosslinks (DPCs). DPCs are a cytotoxic form of DNA damage that potently block replication and transcription. Despite their biological significance, DPCs are often overshadowed by other forms of damage, such as DNA-DNA interstrand crosslinks (ICLs). The development of these molecules could be useful for studying DPC formation in the test tube, in cells and possibly even organisms. Selective DPC formation would enable scientists to study how such cytotoxic lesions are repaired and determine the consequences of their formation on processes involved in genetic expression. These molecules could be especially useful in the complex environment of cells, where the formation of mixtures makes it particularly difficult to elucidate the consequences of individual chemical products. This project will provide students with broad training in several research areas, including synthetic organic chemistry, biochemistry, and cell biology. The students that obtain training while contributing to this project, will include those from groups that have historically been underrepresented in the sciences.Under this award, the Greenberg laboratory will focus synthetic efforts on bis-electrophiles that take advantage of the high reactivity of nitrogen mustards to rapidly alkylate DNA. However, one of the highly reactive chloroethyl groups will be replaced with a more selective electrophile that is designed to preferentially react with the side-chain amino group of lysines. In this way, the formation of monoalkylation products that account for the majority of nitrogen mustard reactivity will be disfavored. Furthermore, the selectivity of the electrophile to form DPCs over ICLs will be determined. Selective DPC formation will be tested in nucleosome core particles, as well as in cells. Preferential DPC formation compared to ICLs will be probed using various cell lines that are deficient in repair of one lesion family. For instance, it is hypothesized that cells lacking SPRTN, a protease that initiates DPC repair, will be hypersensitive to the molecules. Selective DPC formation in cells that lack SPRTN would enable the exploration of alternative DPC repair systems. Studies with these bis-electrophiles may provide the foundation for the discovery of therapeutic agents that act by forming DPCs in targeted cellular loci.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学过程中的化学过程（CLP）方案的支持下，约翰·霍普金斯大学（Johns Hopkins University）的马克·格林伯格（Marc Greenberg）教授及其研究团队正在研究DNA蛋白交联选择性形成分子的发展（DPCS）。 DPC是DNA损伤的一种细胞毒性形式，可有效阻止复制和转录。尽管具有生物学意义，但DPC通常会被其他形式的损害（例如DNA-DNA Interstrand交叉链接（ICL））所掩盖。这些分子的开发对于在测试管，细胞甚至生物体中研究DPC形成可能很有用。选择性DPC的形成将使科学家能够研究如何修复这种细胞毒性病变，并确定其形成对遗传表达涉及过程的后果。这些分子在复杂的细胞环境中可能特别有用，在这些细胞的复杂环境中，混合物的形成使得很难阐明单个化学产物的后果。该项目将为学生提供多个研究领域的广泛培训，包括合成有机化学，生物化学和细胞生物学。在为该项目做出贡献时获得培训的学生将包括从历史上不足以代表的小组的学生。在该奖项之后，格林伯格实验室将集中于合成的努力上，以利用氮芥末酱的高反应性来快速烷基烷基DNA。但是，高反应性的氯乙基基团之一将被更具选择性的电力释放量代替，该基因旨在优先与侧链氨基赖氨酸组反应。以这种方式，将不利的是氮芥末反应性的大部分氮烷基化产物的形成。此外，将确定亲电的选择性在ICL上形成DPC。选择性DPC形成将在核小体核心颗粒以及细胞中进行测试。与ICL相比，将使用一个病变家族修复的各种细胞系进行探测优先的DPC形成。例如，假设缺乏SPRTN的细胞（一种启动DPC修复的蛋白酶）对分子非常敏感。缺乏SPRTN的细胞中的选择性DPC形成将使替代DPC修复系统探索。对这些双遗传噬菌体的研究可能为发现通过在靶向细胞基因座中形成DPC来发现的治疗剂的基础。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛影响的审查标准通过评估来通过评估来获得支持的。