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

基本信息

批准号：
7338346
负责人：
SANDRA K WELLER
金额：
$ 28.2万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
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/7338346
关键词：
Active Sites Address Affect Antiviral Agents Antiviral Therapy Biochemical Biological Biological Assay Cancer Biology Cell Cycle Checkpoint Cell Line Cell physiology Cells Complex Condition DNA DNA Binding DNA Damage DNA Sequence Rearrangement DNA biosynthesis DNA chemical synthesis Defect Defective Viruses Development Disease Endopeptidases Filament Fluorescence Frequencies Genetic Genetic Recombination Genome Genome Stability Genomics Goals Growth Herpesviridae Herpesvirus 1 Human Immunocompetent Immunocompromised Host In Vitro Individual Infection Knock-out Lead Lesion Malignant Neoplasms Mammalian Cell Maps Measures Methods Monitor Mutation Pathway interactions Patients Peptide Hydrolases Plasmids Process Proteins Proteolysis Pulsed-Field Gel Electrophoresis Recruitment Activity Resistance Roentgen Rays Role SS DNA BP Side Simplexvirus Site Small Interfering RNA Spectrum Analysis Stress Structure Testing Transfection Viral Viral Proteins Virus Virus Diseases Work analytical ultracentrifugation base design fascinate gene replacement homologous recombination improved in vivo mutant neonate pathogen prevent recombinase recombinational repair repaired response spleen exonuclease stoichiometry

项目摘要

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.

单纯疱疹病毒（HSV-1）是负责自限制性粘膜状的重要人类病原体免疫能力患者的病变以及新生儿和其他其他潜在致命感染免疫功能低下的个体。在此提案中，我们将测试HSV使用的假设重组依赖性复制以使用病毒和细胞蛋白同时复制其基因组。我们提出HSV已经演变为与细胞修复和重组机制相互作用细胞通常用于响应DMADAMAGE和其他应力因素。旨在监测和修复受损的DMA对于维持基因组稳定性至关重要。哺乳动物细胞暴露 DNA损坏的药物诱导细胞周期检查点和DMA修复途径，以保护来自突变和基因组重排的细胞。这些途径中的缺陷导致疾病癌症。疱疹病毒与他们的宿主共同开发，并开发了令人着迷的副步进方式，颠覆，在某些情况下受益于宿主细胞反应。在此提案中，我们将测试 HSV使用同源重组（HR）修复途径的假设是为了自身的利益。我们的初步工作表明，HSV使用病毒和细胞蛋白的组合进行产生后代基因组所需的重组依赖性复制。我们以前有证明病毒5'至3'核酸外切酶，UL12和主要的单链DNA结合蛋白， ICP8可以充当两亚基重组酶。我们建议继续研究这种病毒重组酶并检验了HSV病毒和细胞复制基因组的假设。 AIM 1WILL检验重组对于生产性病毒感染至关重要的假设； AIM 2将测试假设ICP8和UL12在感染过程中共同起作用； AIM 3将检验以下假设细胞重组蛋白被募集以与病毒预性位点并相互作用复制中心。遗传，生物物理，生化和细胞生物学方法的结合将被使用。 HSV-1是一种主要的人类病原体，对DNA复制机制知之甚少。我们有提议该病毒利用细胞用于预防遗传不稳定的一些机械和癌症。由于DNA复制是抗病毒药物的主要靶标，因此必须充分理解很重要在此过程中的关键参与者是为了制定更好的治疗策略。在这个建议中，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。