Viral and Host Determinants of Parvovirus Replication

细小病毒复制的病毒和宿主决定因素

基本信息

批准号：
10534743
负责人：
Jianming Qiu
金额：
$ 47.21万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-27 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10534743
关键词：
Acute Air Antiviral Agents Autonomous Replication Capsid Proteins Cell Cycle Cell Nucleus Cell model Cells Child Circovirus Complex DNA Damage DNA Repair DNA Replication Factor DNA Viruses DNA biosynthesis DNA replication origin DNA-Directed DNA Polymerase DNA-PKcs Dependence Dependovirus Double-Stranded RNA Embryo Epithelial Cells Family G2 Phase Arrest Gene Delivery Gene Expression Genetic Transcription Genome Goals Hela Cells Helper Viruses Human In Vitro Infection Interphase Cell KRP protein Kidney Knowledge Lead Licensing Liquid substance Mediating Modeling Nonstructural Protein Nucleotides Open Reading Frames PRKR gene Papillomavirus Parvovirus Parvovirus Infections Pathogenicity Pathway interactions Phosphatidylinositols Play Polymerase Process Production Productivity Property Proteins Proteomics RNA Polymerase II RNA Polymerase III RNA metabolism Rad30 protein Recombinant adeno-associated virus (rAAV)Regulation Respiratory Tract Infections Role S phase Signal Transduction Small RNA Testing Transfection Translations Untranslated RNA Viral Viral Genes Viral Genome Viral Nonstructural Proteins Virus Diseases Virus Replication adeno-associated viral vector airway epithelium ataxia telangiectasia mutated protein clinically relevant delivery vehicle gene therapy in vivo mRNA Precursor mRNA Translation novel postmitotic prevent protein expression protein kinase R recruit response therapeutic development viral DNA

项目摘要

Human bocavirus 1 (HBoV1), an autonomous human parvovirus, causes acute respiratory tract infections in young children. HBoV1 is unique among all small DNA viruses in that it expresses a noncoding RNA of 140 nucleotides from the 3' noncoding region after the capsid protein-coding region. This bocavirus-encoded small RNA (BocaSR) is exclusively expressed in the nucleus and plays an important role both in expression of viral nonstructural proteins and in viral DNA replication. HBoV1 infects and replicates in terminally-differentiated human airway epithelium cultured at an air-liquid interface (HAE-ALI), mimicking natural infection of human airways. Importantly, HBoV1 genome replication in the nondividing airway epithelial cells utilizes error-free Y- family DNA repair DNA polymerase (Pol) η and Pol κ, and BocaSR specifically interacts with viral nonstructural proteins. On the other hand, HBoV1 is a helper virus for the replication of adeno-associated virus (AAV) in HAE-ALI, and BocaSR also plays a key role in facilitating AAV Rep gene expression and DNA replication. Our central hypothesis is that HBoV1 has evolved to express a noncoding RNA to facilitate expression of the viral nonstructural proteins and their function in licensing viral DNA replication, which mimics a DNA repair process driven by error-free Pol η and Pol κ. In this application, we aim to reveal the mechanisms underlying the BocaSR-regulated expression of viral nonstructural proteins and viral DNA replication in the nucleus, and to understand the viral manipulation of the error-free Pol η and Pol κ in DNA synthesis. The BocaSR is novel as it is the first one found among all small DNA viruses and differs from other known RNA polymerase III (Pol III) transcribed viral noncoding RNAs by its unique properties in regulating viral gene expression and playing a direct role in viral DNA replication in the nucleus. Studying BocaSR-regulating parvovirus replication will contribute to the understanding of the functions of other viral noncoding RNAs in virus replication. In addition, knowledge in how parvovirus employs the DNA repair machinery for viral genome replication in nondividing cells will help us understand how a small DNA virus overcomes the barrier to replicate its genome outside of the cell cycle. Dissecting the viral and host determinants of HBoV1 genome replication will help the development of therapeutic approaches to prevent acute respiratory tract infections caused by HBoV1. Finally, deep understanding AAV DNA replication will help find a better way to produce recombinant AAV (rAAV) vector and to increase the efficacy of rAAV gene delivery, in particular, to nondividing cells.

人类博卡病毒 1 (HBoV1) 是一种自主性人类细小病毒，可引起幼儿急性呼吸道感染。 HBoV1 在所有小 DNA 病毒中是独一无二的，因为它表达衣壳蛋白后 3' 非编码区的 140 个核苷酸的非编码 RNA。这种博卡病毒编码的小RNA (BocaSR) 仅在细胞核中表达，在病毒非结构蛋白的表达和病毒中发挥着重要作用。 DNA 复制在气-液界面培养的终末分化的人气道上皮 (HAE-ALI) 中进行感染和复制，模仿人气道的自然感染，重要的是，HBoV1 基因组在非分裂气道上皮细胞中的复制利用无错误的 Y。 - 家族 DNA 修复 DNA 聚合酶 (Pol) η 和 Pol κ，以及 BocaSR 与病毒非结构蛋白特异性相互作用。 HBoV1 是 HAE-ALI 中腺相关病毒 (AAV) 复制的辅助病毒，BocaSR 在促进 AAV Rep 基因表达和 DNA 复制方面也发挥着关键作用。我们的中心假设是 HBoV1 已进化为表达非编码蛋白。 RNA 促进病毒非结构蛋白的表达及其在许可病毒 DNA 复制中的功能，模拟由无错误 Pol η 和 Pol κ 驱动的 DNA 修复过程。在本应用中，我们旨在揭示其背后的机制。 BocaSR 调节病毒非结构蛋白的表达和病毒在细胞核中的 DNA 复制，并了解 DNA 合成中无差错 Pol η 和 Pol κ 的病毒操作。 BocaSR 是新颖的，因为它是所有小型病毒中第一个发现的。 DNA 病毒与其他已知的 RNA 聚合酶 III (Pol III) 转录的病毒非编码 RNA 不同，它具有调节病毒基因表达的独特特性，并在细胞核中的病毒 DNA 复制中发挥直接作用。将有助于了解其他病毒非编码RNA在病毒复制中的功能。此外，关于细小病毒如何利用DNA修复机制在非分裂细胞中进行病毒基因组复制的知识将有助于我们了解小型DNA病毒如何克服复制障碍。剖析 HBoV1 基因组复制的病毒和宿主决定因素将有助于开发预防 HBoV1 引起的急性呼吸道感染的治疗方法。最后，深入了解 AAV DNA 复制将有助于找到更好的方法。生产重组 AAV (rAAV) 载体并提高 rAAV 基因递送的效率，特别是递送至非分裂细胞的效率。