Investigating the interaction between DNA-PK, cGAS and IFI16 in the innate immune response pathway

研究先天免疫反应途径中 DNA-PK、cGAS 和 IFI16 之间的相互作用

• 批准号: 2737806
• 金额: --
• 依托单位: University of Leicester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2737806
• 关键词: Investigating; interaction; between; DNA; PK

DNA-dependent protein kinase (DNA-PK) is a complex of proteins that acts as a crucial component of the non-homologous end joining (NHEJ) mechanism of DNA double-strand break repair. However, recent studies suggest that DNA-PK may have an additional secondary role in regulating the innate immune response to viral infections. These studies indicate that DNA-PK may regulate the type 1 interferon response by interacting with cGAS and through the cGAS interacting with IFI16, leading to the STING pathway. The experimental evidence of interactions between cGAS and DNA-PK as well as between IFI16 and cGAS suggests that both IFI16 and DNA-PK may be important regulators of the type 1 interferon response. This PhD project will aim to elucidate whether DNA-PK and/or IFI16 regulate the cGAS-STING pathway, by examining whether there are direct interactions between these proteins. This will involve protein expression, purification and biochemical/biophysical techniques to determine protein and DNA interactions. Through this experimental work, this project will help determine whether DNA-PK or IFI16 bind to cGAS and, if both bind, whether these proteins bind to cGAS simultaneously to form a single complex. Ultimately, cryo-electron microscopy will be utilised to study the structure of these complexes and determine their specific interactions. This work will help establish whether these proteins regulate the innate immune response to viruses and will provide an indication of the mechanism through which this occurs. Consequently, this project will determine the effect that DNA-PK and IFI16 have on the intensity/efficacy of viral clearance within infected host cells. Furthermore, understanding these interactions and the mechanism will help guide development of future vaccines and antivirals to enable new strategies for combating viral infections.

DNA依赖性蛋白激酶（DNA-PK）是蛋白质的复合物，它是DNA双链破裂修复的非同源末端连接（NHEJ）机制的关键成分。但是，最近的研究表明，DNA-PK在调节对病毒感染的先天免疫反应中可能具有额外的二级作用。这些研究表明，DNA-PK可以通过与CGA相互作用以及通过与IFI16相互作用的CGA来调节1型干扰素响应，从而导致STING途径。 CGA与DNA-PK以及IFI16和CGA之间相互作用的实验证据表明，IFI16和DNA-PK都可能是1型干扰素反应的重要调节剂。该博士学位项目将旨在通过检查这些蛋白之间是否存在直接相互作用来阐明DNA-PK和/或IFI16是否调节CGAS-Sting途径。这将涉及蛋白质表达，纯化和生化/生物物理技术，以确定蛋白质和DNA相互作用。通过这项实验性工作，该项目将有助于确定DNA-PK或IFI16是否与CGA结合，并且是否两者结合，这些蛋白是否同时与CGA结合以形成单个复合物。最终，将使用冷冻电子显微镜来研究这些复合物的结构并确定它们的特定相互作用。这项工作将有助于确定这些蛋白质是否调节对病毒的先天免疫反应，并提供发生这种情况的机制。因此，该项目将确定DNA-PK和IFI16对感染宿主细胞内病毒清除的强度/功效的影响。此外，了解这些相互作用以及该机制将有助于指导未来的疫苗和抗病毒药物的开发，以实现对抗病毒感染的新策略。