Molecular characterization of the bocaviruses

博卡病毒的分子特征

• 批准号: 8067896
• 负责人: Jianming Qiu
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8067896
• 关键词: Acute; Animals; Apoptotic; Breathing; Canis familiaris; Caspase; Cell Cycle Arrest; Cell Death; Cells; Child; Clinical; DNA; DNA Damage; DNA biosynthesis; Diarrhea; Disease; Epithelial Cells; Family; Genome; Host Defense Mechanism; Human; Human Cloning; Infection; Knowledge; Lower Respiratory Tract Infection; Lung diseases; Mitosis; Molecular; Open Reading Frames; Parvoviridae; Parvovirus; Pathogenesis; Pneumonia; Process; Proteins; Role; S Phase; Specimen; Symptoms; Therapeutic Intervention; Viral; Viral Proteins; Viral Structural Proteins; Viral load measurement; Virus; Virus Diseases; Wheezing; base; biological systems; bovine parvovirus; design; enteritis; genome sequencing; human disease; insight; novel; prevent; prophylactic; public health relevance; pup; research study; respiratory; response; virus pathogenesis

DESCRIPTION (provided by applicant): Human bocavirus (HBoV) is associated with acute expiratory wheezing and pneumonia. The HBoV genome has been frequently detected worldwide, ranging from 2%-19%, in respiratory specimens from young children with acute respiratory illnesses. Differentiated human airway epithelial cells have been recently shown to support HBoV replication; however, a high virus load is required. An infectious DNA clone of HBoV has not been constructed, which impairs study of the pathogenesis of this virus. Minute virus of canines (MVC), a canine bocavirus, causes respiratory diseases with breathing difficulty and enteritis with severe diarrhea in pups. MVC infection of permissive WRD cells perturbs cell progression at the S-phase in favor of viral replication in early infection and at the G2/M-phase in later infection, and ultimately leads to an apoptotic cell death. In addition, a DNA damage response is detected during MVC infection. Therefore, cell death induced by MVC infection, which is independent of both nonstructural and structural viral proteins, must be a host defense mechanism, presumably the DNA damage response, designed to arrest viral replication by eliminating infected cells, and must be regulated. In addition, bocaviruses are unique among parvoviruses in that a third ORF, the mid-ORF, encodes a novel non-structural protein, NP1. Because of the similarities between HBoV and MVC, both in the organization and sequence of the genomes and in the symptoms of caused diseases, study of the canine Bocavirus MVC will eventually answer questions about the functions of NP1 in Bocavirus DNA replication and the molecular basis of Bocavirus-induced cell cycle arrest and cell death. Ultimately, knowledge obtained from study of MVC will help us to understand the molecular pathogenesis of HBoV. In this application, we propose to discover the function of the NP1 protein in Bocavirus DNA replication, especially, to identify cellular factors that interact with the NP1; and how Bocavirus infection arrests the cell cycle and causes an apoptotic cell death; and attempt to construct an infectious clone of HBoV from clinical specimens. In addition, the strategy that Bocavirus infection uses to induce a DNA damage response, leading to cell cycle arrest and cell death, is unusual in that viral proteins are not involved. Examining this strategy within the tractable biological system of Bocavirus provides us an attractive opportunity to gain insight into the basic cellular mechanism involved in defending against viral infection. PUBLIC HEALTH RELEVANCE: Human bocavirus is pathogenic to humans, causes lower respiratory tract infections in children. Study of the animal bocavirus will help us to understand the replication and pathogenesis of the bocaviruses in general. Our experiments will thus answer critical questions in the pathogenesis of human bocavirus infection, and have the potential to provide prophylactic and therapeutic interventions to diseases caused by human bocavirus.

描述(申请人提供)：人类博卡病毒(HBoV)与急性呼气性喘息和肺炎有关。HBoV基因组在世界范围内经常被检测到，从患有急性呼吸道疾病的幼儿的呼吸道样本中检测到的比例从2%到19%不等。分化的人呼吸道上皮细胞最近被证明支持HBoV复制；然而，需要高病毒载量。HBoV的感染性DNA克隆尚未构建，这给HBoV致病机理的研究带来了一定的困难。犬细小病毒(MVC)是一种犬博卡病毒，可引起仔犬呼吸困难的呼吸系统疾病和严重腹泻的肠炎。MVC感染WRD细胞后，细胞处于S期，早期感染时有利于病毒复制，晚期感染时处于G2/M期，最终导致细胞凋亡。此外，在MVC感染期间还检测到DNA损伤反应。因此，MVC感染诱导的细胞死亡必须是一种宿主防御机制，可能是DNA损伤反应，旨在通过清除受感染的细胞来阻止病毒复制，并且必须受到调控。此外，博卡病毒在细小病毒中是独一无二的，因为第三个ORF，MID-ORF编码一种新的非结构蛋白NP1。由于HBoV和MVC在基因组的组织和序列以及所引起的疾病症状上的相似性，对犬博卡病毒MVC的研究最终将回答关于NP1在博卡病毒DNA复制中的功能以及博卡病毒诱导细胞周期停滞和细胞死亡的分子基础的问题。最终，从MVC研究中获得的知识将有助于我们理解HBoV的分子发病机制。在这一应用中，我们建议发现NP1蛋白在博卡病毒DNA复制中的功能，特别是识别与NP1相互作用的细胞因子；以及博卡病毒感染如何阻止细胞周期和导致细胞凋亡；并试图从临床标本中构建具有感染性的HBoV克隆。此外，博卡病毒感染用来诱导DNA损伤反应，导致细胞周期停滞和细胞死亡的策略是不寻常的，因为病毒蛋白不参与。在博卡病毒易受控制的生物系统中研究这一策略为我们提供了一个有吸引力的机会，让我们深入了解防御病毒感染的基本细胞机制。 与公共卫生相关：人类博卡病毒对人类是致病的，会导致儿童下呼吸道感染。对动物博卡病毒的研究将有助于我们从总体上了解博卡病毒的复制和致病机制。因此，我们的实验将回答人类博卡病毒感染发病机制中的关键问题，并有可能为人类博卡病毒引起的疾病提供预防和治疗干预。