Influenza Virus Assembly

流感病毒组装

基本信息

批准号：
7617137
负责人：
YOSHIHIRO KAWAOKA
金额：
$ 40.42万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-05-01 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7617137
关键词：
Affect American Antiviral Agents Attenuated Vaccines Bioterrorism Birds Categories Classification Communicable Diseases Complementary DNA Complex Cytoplasmic Tail Development Electrons Emerging Communicable Diseases Event Family suidae Fibrinogen Generations Genetic Techniques Genome Goals Hemagglutinin Human Human Virus Individual Infectious Diseases Research Influenza Integral Membrane Protein Intervention Ion Channel Protein Laboratories Life Cycle Stages Maintenance Mediating Mediator of activation protein Microscopic Modeling Molecular National Institute of Allergy and Infectious Disease Neuraminidase Nucleotides Plasmids Play Polymerase Population Proteins RNA Reassortant Viruses Research Research Personnel Ribonucleoproteins Role Signal Transduction System Technology Testing Viral Viral Genome Viral Structural Proteins Virion Virulent Virus Virus Assembly base biodefense combat design influenza virus strain influenzavirus insight killings macromolecule novel pandemic disease pandemic influenza particle pathogen positional cloning programs research study viral RNA

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand the assembly of influenza viruses at the molecular level. To this end, we propose to elucidate mechanism(s) by which influenza virus RNA segments are incorporated into virions, and to assess the contribution of selective viral (v) RNA incorporation to the generation of new reassortant influenza viruses. A plasmid-based reverse genetics system, developed in the applicant's laboratory, allows influenza virus to be produced entirely from cloned cDNA. This technology was used to identify segment-specific regions that mediate vRNA virion incorporation, providing the first direct evidence for a selective rather than random mechanism of vRNA packaging. Aim 1 seeks to refine the packaging signals within these regions, while Aim 2 determines if interactions among these signals mediate the assembly of sets of eight different vRNA segments. Aim 3 attempts to elucidate the mechanism(s) by which these sets of vRNA segments are incorporated into budding virions. The virion incorporation of vRNAs appears to involve viral structural proteins that sequester the viral genome segments into budding particles. Candidate mediators of this interaction (from the virion shell) are the three viral transmembrane proteins HA, NA, and ion channel protein (M2), which may execute this function through their cytoplasmic tails; candidates, from the viral ribonucleoprotein complex associated with each vRNA segment include the three polymerase proteins (PB2, PBI, PA). Aim 4 will test the hypothesis that selective vRNP incorporation plays a significant role in the reassortment and generation of new influenza virus strains. Indeed, the 1957 and 1968 pandemic influenza strains, as well as a triple reassortant that appeared in the North American pig population in 1998, were characterized by the introduction of a PB1 segment, in addition to HA and NA segments or an HA segment, into a new host. Thus, interactions between the PB1 vRNA and other vRNA segments (e.g., HA) may govern their preferential incorporation into virions. The proposed experiments will provide new fundamental insights into the life cycle of influenza viruses, in particular the mechanisms for organized recruitment of multiple vRNA segments into budding virions. If selective vRNA incorporation contributes significantly to the generation of novel reassortant viruses, new avenues will be opened for the development of antiviral interventions, including compounds that interfere with efficient vRNA virion incorporation or perhaps live vaccines that do not reassort with field strains due to altered incorporation sequences.

描述（由申请人提供）：这项研究的长期目标是了解分子水平上流感病毒的组装。为此，我们建议阐明将流感病毒RNA片段纳入病毒体的机制，并评估选择性病毒（V）RNA掺入对新的重构流感病毒的产生的贡献。在申请人实验室中开发的基于质粒的反向遗传学系统使流感病毒完全由克隆的cDNA产生。该技术用于识别介导VRNA病毒体掺入的细分市场特异性区域，为VRNA包装的选择性而不是随机机理提供了第一个直接证据。 AIM 1试图完善这些区域内的包装信号，而AIM 2决定了这些信号之间的相互作用是否介导了八个不同VRNA段的集合组装。 AIM 3尝试阐明这些VRNA段集合到萌芽病毒体中的机制。 VRNA的病毒体掺入似乎涉及病毒结构蛋白，这些蛋白将病毒基因组片段隔离到萌芽颗粒中。这种相互作用的候选介质（来自Virion壳）是三个病毒跨膜蛋白HA，Na和Ion通道蛋白（M2），可以通过其细胞质尾巴执行该功能。候选物来自与每个VRNA相关的病毒核糖核蛋白复合蛋白复合物，包括三个聚合酶蛋白（PB2，PBI，PBI，PA）。 AIM 4将检验以下假设：选择性VRNP融合在新型流感病毒菌株的重新分类和生成中起重要作用。的确，1957年和1968年的大流行性流感菌株以及1998年出现在北美猪种群中的三重重新分类，其特征在于引入了PB1段，除了HA和NA片段或HA段或HA段，还引入了新宿主。因此，PB1 VRNA与其他VRNA片段（例如HA）之间的相互作用可以控制其优先掺入病毒体中。提出的实验将为流感病毒的生命周期提供新的基本见解，特别是将多个VRNA片段组织为崭露头角的病毒体的机制。如果选择性VRNA融合有助于产生新的重新分类病毒，则将开放新的途径，以开发抗病毒干预措施，包括干扰有效的VRNA Virion掺入或可能因综合序列而导致的现场菌株而不会将现场菌株重新培养的化合物。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Strand-specific real-time RT-PCR for distinguishing influenza vRNA, cRNA, and mRNA.

DOI：
10.1016/j.jviromet.2010.12.014
发表时间：
2011-04
期刊：
JOURNAL OF VIROLOGICAL METHODS
影响因子：
3.1
作者：
Kawakami, Eiryo;Watanabe, Tokiko;Fujii, Ken;Goto, Hideo;Watanabe, Shinji;Noda, Takeshi;Kawaoka, Yoshihiro
通讯作者：
Kawaoka, Yoshihiro

Cellular networks involved in the influenza virus life cycle.