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：
10311526
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/10311526
关键词：
Acute Air Antiviral Agents 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 Translation Genome Goals Hela Cells Helper Viruses Human In Vitro Infection Interphase Cell KRP protein Kidney Knowledge Lead Licensing Liquid substance Mediating Mitotic Modeling Nonstructural Protein Nucleotides Open Reading Frames Papillomavirus Parvovirus Parvovirus Infections Pathogenicity Pathway interactions Phosphatidylinositols Phosphorylation Play Polymerase Process Production 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 Translations Untranslated RNA Viral Viral Genes Viral Genome Viral Nonstructural Proteins Virus Diseases Virus Replication adeno-associated viral vector airway epithelium clinically relevant delivery vehicle gene therapy in vivo mRNA Precursor novel 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Bocavirus 1（HBOV1）是一种自主的人细节病毒，会引起幼儿急性呼吸道感染。 HBOV1在所有小型DNA病毒中都是独一无二的，因为它表达了在衣壳蛋白编码区域之后3'非编码区域的140个核动脉介质的非编码RNA。该Bocavirus编码的小RNA（BOCASR）仅在细胞核中表达，并在病毒非结构蛋白的表达和病毒DNA复制中起着重要作用。 HBOV1感染和复制在空气界面（HAE-ALI）培养的末端分化的人类气道上皮，模仿了人类气道的自然感染。重要的是，非各个气道上皮细胞中的HBOV1基因组复制利用无误差Y家族DNA修复DNA聚合酶（POL）η和polκ，BOCASR与病毒非结构性蛋白质特异性相互作用。另一方面，HBOV1是用于在HAE-ALI中复制腺相关病毒（AAV）的辅助病毒，BOCASR在支持AAV REP基因表达和DNA复制方面也起着关键作用。我们的中心假设是，HBOV1已演变为表达非编码RNA，以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，该复制模仿了由无错误的polη和polκ造成的DNA修复过程。在此应用中，我们旨在揭示BOCASR调节的病毒非结构蛋白表达和病毒DNA在细胞核中的机制，并了解DNA合成中无误差POL或POLκ的病毒操纵。 BOCASR是新颖的，因为它是所有小型DNA病毒中发现的第一个，并且与其他已知的RNA聚合酶III（POL III）不同，它通过其在调节病毒基因表达中的独特特性转录了病毒非编码RNA，并且在核中病毒DNA重复中起着直接的作用。研究BOCASR调节细胞小旋转病毒的复制将有助于理解其他病毒非编码RNA在病毒复制中的功能。此外，了解细小病毒雇员如何在非分裂细胞中进行病毒基因组复制的DNA修复机制如何帮助我们了解小的DNA病毒如何克服障碍物在细胞周期之外复制其基因组的障碍。解剖HBOV1基因组复制的病毒和宿主决定剂将有助于开发治疗方法，以防止HBOV1引起的急性呼吸道感染。最后，深入了解AAV DNA复制将有助于找到一种更好的方法来产生重组AAV（RAAV）载体，并提高RAAV基因递送的效率，尤其是对非分散细胞的效率。