FUNCTIONS OF THE PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS

人呼吸道合胞病毒蛋白质的功能

• 批准号: 6288863
• 负责人: PETER LEON COLLINS
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6288863
• 关键词: DNA directed RNA polymerase; RNA biosynthesis; genetic transcription; microorganism culture; microorganism growth; nucleocapsid; phosphoproteins; plasmids; posttranslational modifications; protein structure function; replicase; respiratory syncytial virus; tissue /cell culture; transfection; virus RNA; virus envelope; virus genetics; virus protein; virus replication

Human respiratory syncytial virus (RSV) is an enveloped RNA-containing virus of the paramyxovirus family of Order Mononegavirales (the nonsegmented negative strand RNA viruses). RSV is the most important viral agent of pediatric respiratory tract disease worldwide. Other members of the Order include measles, mumps, rabies and Ebola viruses. The RSV genome is a single negative strand of RNA of 15,222 nucleotides that encodes 10 major mRNAs and 11 proteins, as we have shown previously. The purpose of this project is to identify the functions of the viral proteins and to reconstruct events in the viral growth cycle under conditions where they can be more readily studied. Knowledge of the functions of the proteins is important for a basic understanding of this important human pathogen and for the design of live-attenuated recombinant vaccine viruses.We previously developed an experimental system for RSV based on the intracellular coexpression and assembly of viral RNA and protein components from recombinant plasmid vectors. This involves transfecting cultured cells with plasmids which individually encode a truncated version of negative-sense genomic or positive-sense antigenomic RNA (the latter being an intermediate in RNA replication) as well as whatever mix of RSV proteins is desired. In this way, various steps in the viral replication cycle can be reconstituted, such as genome replication, gene expression, gene regulation, packaging and infection. Because each component is expressed individually, each can be manipulated individually with regard to relative amount as well as by mutagenesis for structure-function studies. - virus, respiratory tract disease, infectious disease, pediatrics, vaccines, live- attenuated viral vaccine, recombinant DNA, proteins

人呼吸道合胞病毒(RSV)是单毒目副粘病毒家族(非分段负链RNA病毒)的一种囊膜RNA病毒。呼吸道合胞病毒是世界范围内引起儿童呼吸道疾病的最重要的病毒物。该组织的其他成员包括麻疹、腮腺炎、狂犬病和埃博拉病毒。如前所述，RSV基因组是由15,222个核苷酸组成的单条负链RNA，编码10个主要的mRNAs和11个蛋白质。这个项目的目的是确定病毒蛋白的功能，并在更容易研究它们的条件下重建病毒生长周期中的事件。了解这些蛋白质的功能对于基本了解这种重要的人类病原体和设计减毒活疫苗病毒非常重要。我们之前开发了一个基于重组质粒载体中病毒RNA和蛋白质组分的细胞内共表达和组装的RSV实验系统。这涉及到用分别编码截短的负义基因组或正义反基因组RNA(后者是RNA复制的中间产物)以及所需的RSV蛋白质的任何混合的质粒来转染培养的细胞。通过这种方式，病毒复制周期的各个步骤可以被重构，如基因组复制、基因表达、基因调控、包装和感染。因为每一组分都是单独表达的，所以每一组分都可以在相对数量上单独操作，也可以通过突变来进行结构功能研究。-病毒、呼吸道疾病、传染病、儿科、疫苗、减毒活疫苗、重组DNA、蛋白质