Single molecule studies of the interplay between DNA repair mechanisms and between DNA repair and DNA replication and transcription

DNA 修复机制之间以及 DNA 修复与 DNA 复制和转录之间相互作用的单分子研究

• 批准号: 508046170
• 负责人: Dr. Ingrid Teßmer, Ph.D.
• 金额: --
• 依托单位: Rudolf-Virchow-Zentrum - Center for Integrative and Translational Bioimaging
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/508046170?language=en
• 关键词: Single; molecule; studies; interplay; between

The stability of DNA is constantly chemically and physically threatened, resulting in structurally altered or chemically modified DNA that can lead to cancer, cell senescence, or apoptosis. Different DNA modifications are repaired by a number of DNA repair mechanisms that are fine-tuned to their particular target lesions. Several recent studies have also pointed at interactions between different DNA repair systems as well as between DNA repair and DNA transcription and replication. We plan to investigate the interplay of direct damage reversal DNA alkyltransferases and base excision repair (BER) glycosylases with proteins from the nucleotide excision repair (NER) system. Initiation of NER by alkyltransferases and glycosylases may be achieved via stalling of transcription or direct protein-protein interactions in transcription-coupled NER or global genome NER, respectively. Alkyltransferases have also been shown to interact with proteins involved in the initiation of DNA replication. Our single molecule imaging approaches using atomic force microscopy (AFM) and fluorescence optical tweezers, combined with on the one hand ensemble biophysical and functional biochemical assays and on the other hand crystallisation studies aim to provide mechanistic insight into the interactions between these fundamentally important DNA processing systems.

DNA 的稳定性不断受到化学和物理威胁，导致 DNA 结构改变或化学修饰，从而导致癌症、细胞衰老或凋亡。不同的 DNA 修饰是通过许多 DNA 修复机制来修复的，这些机制针对特定的目标病变进行了微调。最近的几项研究还指出了不同 DNA 修复系统之间以及 DNA 修复与 DNA 转录和复制之间的相互作用。我们计划研究直接损伤逆转 DNA 烷基转移酶和碱基切除修复 (BER) 糖基化酶与核苷酸切除修复 (NER) 系统中蛋白质的相互作用。烷基转移酶和糖基化酶启动 NER 可以分别通过转录偶联 NER 或全局基因组 NER 中的转录停滞或直接蛋白质-蛋白质相互作用来实现。烷基转移酶也已被证明与参与 DNA 复制起始的蛋白质相互作用。我们的单分子成像方法使用原子力显微镜 (AFM) 和荧光光镊，一方面结合整体生物物理和功能生化测定，另一方面结合结晶研究，旨在提供对这些基本重要的 DNA 处理系统之间相互作用的机制洞察。