Determining how EBV episome maintenance is regulated by TIMELESS function

确定 TIMELESS 功能如何调节 EBV 附加体维持

• 批准号: MR/X009432/1
• 负责人: Jonathan Baxter
• 金额: $ 64.93万
• 依托单位: University of Sussex
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX009432%2F1
• 关键词: Determining; how; EBV; episome; maintenance

Epstein-Barr virus (EBV) causes 200,000 cancers per year and is responsible for the non-cancerous but debilitating disease, glandular fever. EBV infects the B lymphocytes of the immune system resulting in life long latent infection of the cells where the viral DNA is maintained in cells by a small number of viral proteins. One such viral protein is EBNA1 which binds to the viral DNA and promotes its replication and the continued growth of the cells. Understanding how EBNA1 functions will help us understand how its function can be counteracted and virus infected cells eliminated. One cellular factor that appears to be linked to EBNA1 function is the DNA replication protein TIMELESS (TIM). TIM is required to both prevent the process of DNA replication from causing DNA damage and also to promote the repair of DNA damage detected during DNA replication. Since loss of TIM has been observed to cause breakage of the EBV circular DNA genome in latently infected cells, it appears that TIM works to somehow prevent viral DNA breakage. However, it is unknown which of TIMs several functions in DNA replication and repair are involved in EBV maintenance in infected cells. Understanding how TIM functions in EBV DNA maintenance will not only provide us with a better understanding of how this virus manipulates cellular proteins to maintain infection, but may also lead to new clinical strategies or therapies. TIM is known to work with other DNA repair proteins whose function has being successfully targeted in the clinic. If TIM works with these proteins to maintain EBV infection in cells, we maybe able to repurpose these drugs for use in the treatment of EBV-associated diseases.Therefore, in this proposal we will;1. Genetically manipulate TIM protein expression in cells so that the cell expresses different TIM proteins. Individually these proteins lack the ability to carry out specific functions of TIM while allowing other functions to occur. We will use these modified proteins to identify which functions of TIM are required for continued viral infection in cells. 2. Since loss of TIM is associated with DNA breakage of the viral DNA, we will investigate when and where TIM linked DNA damage occurs and how TIM prevents the accumulation of broken DNA.3. Since TIM is associated with a number of DNA repair pathways, we will examine how loss of these and linked DNA repair factors affect EBV infection. This will allow us to describe the broader context of how TIM functions are linked to the wider response to DNA damage in cells.

Epstein-Barr病毒（EBV）每年引起20万次癌症，并导致非癌性但令人衰弱的疾病，腺热。 EBV感染免疫系统的B淋巴细胞，从而导致细胞的长期潜在感染，其中病毒DNA通过少数病毒蛋白维持在细胞中。一种这样的病毒蛋白是EBNA1，它与病毒DNA结合并促进其复制和细胞的持续生长。了解EBNA1的功能将有助于我们了解如何应对其功能，并消除病毒感染的细胞。似乎与EBNA1功能相关的一个细胞因子是DNA复制蛋白永恒（TIM）。需要TIM才能防止DNA复制过程导致DNA损伤，也可以促进在DNA复制过程中检测到的DNA损伤的修复。由于已经观察到TIM丢失会导致潜在感染细胞中EBV圆形DNA基因组的破裂，因此Tim似乎可以以某种方式预防病毒DNA破裂。但是，未知在DNA复制和修复中的几个功能中，哪些功能参与了受感染细胞的EBV维护。了解TIM在EBV DNA维持中的功能如何不仅可以使我们更好地了解该病毒如何操纵细胞蛋白以保持感染，还可能导致新的临床策略或疗法。众所周知，TIM可以与其他DNA修复蛋白合作，其功能已成功地针对诊所。如果Tim与这些蛋白质合作以维持细胞中的EBV感染，我们也许可以将这些药物重新用于治疗与EBV相关疾病的治疗。因此，在此提案中，我们将; 1; 1。遗传操纵细胞中的TIM蛋白表达，使细胞表达不同的TIM蛋白质。这些蛋白质单独缺乏执行TIM特定功能的能力，同时允许其他功能发生。我们将使用这些修饰的蛋白质来识别细胞中持续病毒感染需要哪些TIM的功能。 2。由于TIM的丢失与病毒DNA的DNA断裂有关，因此我们将研究TIM何时及到何时链接DNA损伤以及TIM如何防止DNA损坏的DNA的积累。3。由于TIM与许多DNA修复途径有关，因此我们将研究这些损失和连接的DNA修复因子如何影响EBV感染。这将使我们能够描述TIM功能如何与细胞中DNA损伤的更广泛响应相关的更广泛的背景。