Modulation of herpes simplex virus type 1 genome structure during lytic replication

裂解复制过程中单纯疱疹病毒 1 型基因组结构的调节

基本信息

批准号：
10352710
负责人：
Jill Ann Dembowski
金额：
$ 17.25万
依托单位：
DUQUESNE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-24 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10352710
关键词：
Architecture Binding Proteins Biological Models Blindness Cell Nucleus Cells Cessation of life ChIP-seq DNA DNA Damage DNA Repair DNA Repair Gene DNA Structure DNA Viruses DNA biosynthesis DNA replication fork Disease Double Strand Break Repair Double Stranded DNA Virus Encephalitis Enzymes Event Excision Gene Expression Gene Transduction Agent Genetic Recombination Genetic Transcription Genome Herpesvirus 1 High-Throughput Nucleotide Sequencing Hot Spot Human Immunocompromised Host Infection Late Gene Transcriptions Life Cycle Stages Ligation Location Lytic Phase Maps Mediating Modeling Nucleotides Okazaki fragments Outcome Play Population Process Proteins Protocols documentation RNA Polymerase II Recurrence Regulation Resolution Role Single Strand Break Repair Site Structure Time Topoisomerase Topoisomerase II Topoisomerase Inhibitors Torsion Transcriptional Regulation Viral Viral Genes Viral Genome Viral Vector Virion Virus Virus Diseases Virus Replication genome-wide in vivo innovation insight lytic replication neonate novel novel strategies pathogen recombinational repair repaired response tool viral DNA

项目摘要

Herpes simplex virus type 1 (HSV-1) is a prevalent pathogen that infects the majority of the human population. Much of the virus life cycle occurs in the nucleus of infected cells, where events that occur on the viral genome determine the outcome of infection. However, how the structure of the viral DNA contributes to the regulation of key viral processes is not well understood. Virion HSV-1 DNA enters into host cells containing single strand breaks and single stranded gaps. During viral DNA replication, branched genome structures form, likely as recombination-mediated replication intermediates. We previously demonstrated that distinct groups of cellular DNA repair proteins associate with viral genomes early during infection and after the onset of viral DNA replication. How DNA damage is navigated by viral DNA replication, transcription, and packaging machinery is not understood. Furthermore, how the genome-wide structure of the HSV-1 genome is modified during the temporal progression of infection has not been defined. A major roadblock to previously investigate these questions was the inability to probe genome-wide DNA structure. In this exploratory proposal, we describe our plan to develop approaches to define the structure of the HSV-1 genome throughout productive infection. The aims outlined in this proposal will establish new approaches to map single and double strand breaks on HSV-1 DNA and investigate viral replication and recombination intermediates in vivo. Results will provide greater insight into mechanisms of regulation of HSV-1 transcription, DNA replication, recombination, and repair. Furthermore, tools generated could be adapted to probe HSV-1 genomes before and after reactivation, viral infection in cells deficient for select cellular DNA repair and DNA damage response factors, and genomes of other DNA viruses.

单纯疱疹病毒1型（HSV-1）是一种流行的病原体，可感染大多数人群。病毒生命周期的大部分发生在感染细胞的核中，其中病毒基因组上发生的事件决定了感染的结果。但是，病毒DNA的结构如何促进关键病毒过程的调节。 Virion HSV-1 DNA进入含有单链断裂和单链间隙的宿主细胞。在病毒DNA复制期间，形成分支的基因组结构，可能是重组介导的复制中间体。我们先前证明，在感染期间和病毒DNA复制发作后，不同的细胞DNA修复蛋白与病毒基因组相关。尚不清楚如何通过病毒DNA复制，转录和包装机制导航DNA损伤。此外，尚未定义在感染时间进展过程中如何修饰HSV-1基因组的全基因组结构。以前研究这些问题的主要障碍是无法探测全基因组DNA结构。在这项探索性建议中，我们描述了制定方法来定义整个生产性感染的HSV-1基因组结构的计划。该提案中概述的目的将建立新的方法，以绘制HSV-1 DNA上的单链断裂并研究体内病毒复制和重组中间体。结果将为HSV-1转录，DNA复制，重组和修复的调节机制提供更多的了解。此外，生成的工具可以适应重新激活前后探测HSV-1基因组，缺乏精选细胞DNA修复的细胞中的病毒感染和DNA损伤响应因子以及其他DNA病毒的基因组。