Role of viral and cellular recombination proteins in HSV DNA replication

单纯疱疹病毒（HSV-1）是一种重要的人类病原体， 免疫功能正常患者的病变和新生儿和其他 免疫力低下的人。在本提案中，我们将测试HSV利用 重组依赖性复制，以使用病毒和细胞蛋白复制其基因组。我们 提出HSV已经进化到与细胞修复和重组机制相互作用， 细胞通常用来应对DM损伤和其他应激因素。这些细胞机器被设计成 监测和修复受损的DNA对维持基因组稳定性至关重要。哺乳动物细胞暴露 DNA损伤剂诱导细胞周期检查点和DNA修复途径， 细胞突变和基因组重排。这些途径的缺陷会导致疾病， 癌疱疹病毒已经与它们的宿主共同进化并发展出了迷人的侧步方式， 破坏并在某些情况下受益于宿主细胞反应。在本提案中，我们将测试 HSV使用同源重组（HR）修复途径是为了自身的利益。我们 初步的研究表明，HSV使用病毒和细胞蛋白的组合来执行 这是产生后代基因组所需的重组依赖性复制。我们先前已经 证明了病毒5'至3'核酸外切酶，UL 12和主要的单链DNA结合蛋白， ICP 8可以作为双亚基重组酶发挥作用。我们建议继续研究这种病毒 重组酶，并测试HSV病毒和细胞途径复制其基因组的假设。 目的1将检验重组对于生产性病毒感染是必要的这一假设;目的2将检验重组对于生产性病毒感染是必要的这一假设。 假设ICP 8和UL 12在感染期间一起工作;目标3将检验假设， 细胞重组蛋白被募集到病毒裂解位点并与之相互作用， 复制中心遗传学、生物物理学、生物化学和细胞生物学方法的组合将 被利用 HSV-1是一种主要的人类病原体，对DNA复制的机制知之甚少。我们有 他提出，病毒利用了一些细胞用来防止遗传不稳定性的相同机制 和癌症由于DNA复制是抗病毒药物的主要靶点，因此充分了解 在这一过程中的主要参与者，以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。

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 DMAdamage and other stress factors. The cellular machinery designed to monitor and repair damaged DMA is essential for maintaining genomic stability. Mammalian cells exposed to DNA damaging agents induce cell cycle checkpoints and DMA repair pathwaysthat serve to protect the cell from mutations and genomic rearrangements. Defects in these pathways lead to diseases such as cancer. Herpesviruses have coevolvedwith 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 pathwaysfor replication of its genome. Aim 1will 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.