Role of viral and cellular recombination proteins in HSV DNA replication

病毒和细胞重组蛋白在 HSV DNA 复制中的作用

• 批准号: 7079573
• 负责人: SANDRA K WELLER
• 金额: $ 28.4万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7079573
• 关键词: DNA replication; genome; infection; proteins; recombinase; role

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV-1) is an important human pathogen responsible for self-limiting mucocutaneous lesions in immunocompetent patients and potentially lethal infections in neonates and other immunocompromised individuals. In this proposal we will test the hypothesis that HSV utilizes recombination-dependent replication to replicate its genome using both viral and cellular proteins. We propose that HSV has evolved to interact with the cellular repair and recombination machinery which the cell normally uses to respond to DNA damage and other stress factors. The cellular machinery designed to monitor and repair damaged DNA is essential for maintaining genomic stability. Mammalian cells exposed to DNA damaging agents induce cell cycle checkpoints and DNA repair pathways that serve to protect the cell from mutations and genomic rearrangements. Defects in these pathways lead to diseases such as cancer. Herpesviruses have coevolved with their hosts and developed fascinating ways of side stepping, subverting and in some cases benefiting from host cell responses. In this proposal we will test the hypothesis that HSV uses the homologous recombination (HR) repair pathway for its own benefit. Our preliminary work suggests that HSV uses a combination of viral and cellular proteins to carry out recombination-dependent replication needed to generate progeny genomes. We have previously demonstrated that the viral 5' to 3' exonuclease, UL12 and the major single strand DNA binding protein, ICP8, can function as a two-subunit recombinase. We propose to continue our studies of this viral recombinase and to test the hypothesis that HSV viral and cellular pathways for replication of its genome. Aim 1 will test the hypothesis that recombination is essential for productive viral infection; Aim 2 will test the hypothesis that ICP8 and UL12 work together during infection; and Aim 3 will test the hypothesis that cellular recombination proteins are recruited to and interact with viral preprelicative sites and active replication centers. A combination of genetic, biophysical, biochemical and cell biological approaches will be used. HSV-1 is a major human pathogen, and little is known about the mechanism of DNA replication. We have proposed that the virus utilizes some of the same machinery that cells use for preventing genetic instability and cancer. Since DNA replication is a major target for antiviral drugs, it is important to fully understand the key players in this process in order to develop better strategies for treatment. In this proposal we will test the hypothesis that HSV uses a combination of viral and cellular proteins to carry out DNA replication.

描述（由申请人提供）：单纯疱疹病毒（HSV-1）是一种重要的人类病原体，导致免疫功能正常患者的自限性皮肤粘膜病变以及新生儿和其他免疫功能低下个体的潜在致命感染。在本提案中，我们将测试 HSV 利用重组依赖性复制利用病毒和细胞蛋白复制其基因组的假设。我们认为 HSV 已经进化到与细胞修复和重组机制相互作用，细胞通常用这些机制来应对 DNA 损伤和其他应激因素。旨在监测和修复受损 DNA 的细胞机制对于维持基因组稳定性至关重要。暴露于 DNA 损伤剂的哺乳动物细胞会诱导细胞周期检查点和 DNA 修复途径，从而保护细胞免受突变和基因组重排的影响。这些途径的缺陷会导致癌症等疾病。疱疹病毒与宿主共同进化，并发展出令人着迷的回避、颠覆以及在某些情况下受益于宿主细胞反应的方式。在本提案中，我们将检验 HSV 使用同源重组 (HR) 修复途径来获取自身利益的假设。我们的初步工作表明，HSV 使用病毒和细胞蛋白的组合来进行产生后代基因组所需的重组依赖性复制。我们之前已经证明，病毒 5' 至 3' 核酸外切酶 UL12 和主要单链 DNA 结合蛋白 ICP8 可以发挥双亚基重组酶的作用。我们建议继续研究这种病毒重组酶，并检验 HSV 病毒和细胞复制其基因组途径的假设。目标 1 将检验重组对于有效病毒感染至关重要的假设；目标 2 将检验 ICP8 和 UL12 在感染期间协同工作的假设；目标 3 将检验细胞重组蛋白被招募到病毒预增殖位点和活跃复制中心并与之相互作用的假设。将结合使用遗传、生物物理、生物化学和细胞生物学方法。 HSV-1 是一种主要的人类病原体，人们对 DNA 复制的机制知之甚少。我们提出，病毒利用了细胞预防遗传不稳定性和癌症的一些相同机制。由于 DNA 复制是抗病毒药物的主要靶标，因此充分了解这一过程中的关键参与者非常重要，以便制定更好的治疗策略。在本提案中，我们将测试 HSV 使用病毒和细胞蛋白的组合来进行 DNA 复制的假设。