Molecular Basis for Ebola Virus Pathogenicity

埃博拉病毒致病性的分子基础

• 批准号: 7189848
• 负责人: YOSHIHIRO KAWAOKA
• 金额: $ 34.75万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7189848
• 关键词: Accounting; Address; Animal Model; Antiviral Agents; Biological; Bioterrorism; Categories; Cells; Cellular Membrane; Classification; Complementary DNA; Containment; Development; Disease; Disease Outbreaks; Ebola virus; Event; Family; Filoviridae; Frankfurt-Marburg Syndrome Virus; Generations; Genes; Genetic Transcription; Genome; Glycoproteins; Goals; Human; Infection; Infectious Diseases Research; Intercistronic Region; Left; Life Cycle Stages; Mediating; Membrane Fusion; Modeling; Molecular; Mucins; Mus; Mutation; National Institute of Allergy and Infectious Disease; Nucleocapsid; Pathogenesis; Pathogenicity; Pharmaceutical Preparations; Plasmids; Positioning Attribute; Proteins; RNA; Reading Frames; Regulation; Reporting; Research; Role; Signal Transduction; Structure; System; Terminator Codon; Therapeutic; Thinking; Transcriptional Regulation; Vaccines; Viral; Viral Genome; Viral Hemorrhagic Fevers; Viral Pathogenesis; Viral Proteins; Virion; Virulence; Virus; Virus Diseases; Work; base; biodefense; biosafety level 4 facility; design; in vitro Assay; nonhuman primate; particle; pathogen; positional cloning; programs; research study; virus pathogenesis

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand the pathogenesis of Ebola virus infection at the molecular level. A potential bioterrorism agent, Ebola virus causes severe hemorrhagic disease in humans and is one of the most deadly human pathogens encountered to date, earning classification as a 'Category A Priority Pathogen' in the NIAID Biodefense and Emerging Infectious Disease Research Opportunities Program. Yet, neither therapeutic treatments nor protective vaccines are available to control this often lethal infection. The combination of a reverse genetics system for the generation of Ebola virus entirely from cloned cDNA and access to a biosafety level 4 facility has positioned the applicant to undertake the following specific aims: 1) To understand the regulation of Ebola viral transcription, in which bicistronic minireplicon and reverse genetics strategies will be used to assess the replicative significance of the unique features characterizing the Ebola virus genome, such as overlapping transcriptional stop/start signals and an unusually long intergenic region; 2) To clarify the mechanisms that drive the formation of infectious Ebola virus particles, in which the roles of the VP40 matrix protein and GP glycoprotein in particle formation and budding will be investigated; 3) To define the role of VP24 protein in Ebola viral replication, in which the intracellular localization of VP24, its association with cellular membranes and/or other viral proteins, and its function in virion budding and membrane fusion will be determined; and 4) To identify the factors and mechanisms responsible for the extreme virulence of Ebola virus infection, in which a murine model of Ebola virus infection will be used to screen mutations that may enhance or diminish viral pathogenicity. Moreover, using a wild-type Ebola virus, furin-mediated GP cleavage, or the mucin-like domain in GP, and the secreted glycoprotein, sGP, will also be evaluated for a possible contribution to Ebola virulence. Collectively, these studies will contribute to the deciphering of the molecular basis for the extreme virulence of this pathogen. Ultimately, the information gained could be exploited in the development of antiviral compounds and effective vaccines.

描述（由申请人提供）：本研究的长期目标是在分子水平上了解埃博拉病毒感染的发病机制。埃博拉病毒是一种潜在的生物恐怖主义制剂，可导致人类严重出血性疾病，是迄今为止遇到的最致命的人类病原体之一，在NIAID生物防御和新兴传染病研究机会计划中被列为“A类优先病原体”。然而，无论是治疗性治疗还是保护性疫苗都无法控制这种通常致命的感染。完全从克隆的cDNA产生埃博拉病毒的反向遗传学系统与获得4级生物安全设施相结合，使申请人能够实现以下具体目标：1）了解埃博拉病毒转录的调控，其中双顺反子小复制子和反向遗传学策略将用于评估表征埃博拉病毒基因组的独特特征的复制意义，如重叠的转录终止/起始信号和异常长的基因间隔区; 2）阐明驱动感染性埃博拉病毒颗粒形成的机制，其中将研究VP 40基质蛋白和GP糖蛋白在颗粒形成和出芽中的作用; 3）确定VP 24蛋白在埃博拉病毒复制中的作用，其中VP 24的细胞内定位，其与细胞膜和/或其他病毒蛋白的结合，并确定其在病毒粒子萌芽和膜融合中的功能; 4）确定导致埃博拉病毒感染极端毒力的因素和机制，其中将使用埃博拉病毒感染的小鼠模型来筛选可能增强或减弱的突变病毒致病性。此外，使用野生型埃博拉病毒，弗林蛋白酶介导的GP切割，或GP中的粘蛋白样结构域，和分泌的糖蛋白，sGP，也将被评估为埃博拉病毒毒力的可能贡献。总的来说，这些研究将有助于破译这种病原体极端毒力的分子基础。最终，获得的信息可以用于开发抗病毒化合物和有效的疫苗。

项目成果

Identification of Amino Acids in Marburg Virus VP40 That Are Important for Virus-Like Particle Budding

• DOI: 10.1093/infdis/jir309
• 发表时间: 2011-11-01
• 期刊: JOURNAL OF INFECTIOUS DISEASES
• 影响因子: 6.4
• 作者: Makino, Akiko;Yamayoshi, Seiya;Kawaoka, Yoshihiro
• 通讯作者: Kawaoka, Yoshihiro

Ebolavirus is internalized into host cells via macropinocytosis in a viral glycoprotein-dependent manner.

• DOI: 10.1371/journal.ppat.1001121
• 发表时间: 2010-09-23
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Nanbo A;Imai M;Watanabe S;Noda T;Takahashi K;Neumann G;Halfmann P;Kawaoka Y
• 通讯作者: Kawaoka Y

Characterization of the Ebola virus nucleoprotein-RNA complex.

埃博拉病毒核蛋白-RNA复合物的表征。

• DOI: 10.1099/vir.0.019794-0
• 发表时间: 2010-06
• 期刊: JOURNAL OF GENERAL VIROLOGY
• 影响因子: 3.8
• 作者: Noda, Takeshi;Hagiwara, Kyoji;Sagara, Hiroshi;Kawaoka, Yoshihiro
• 通讯作者: Kawaoka, Yoshihiro

Ebola virus (EBOV) VP24 inhibits transcription and replication of the EBOV genome.

埃博拉病毒 (EBOV) VP24 抑制 EBOV 基因组的转录和复制。

• DOI: 10.1086/520582
• 发表时间: 2007
• 期刊: The Journal of infectious diseases
• 作者: Watanabe,Shinji;Noda,Takeshi;Halfmann,Peter;Jasenosky,Luke;Kawaoka,Yoshihiro
• 通讯作者: Kawaoka,Yoshihiro

Ebola virus matrix protein VP40 uses the COPII transport system for its intracellular transport.

• DOI: 10.1016/j.chom.2008.02.001
• 发表时间: 2008-03
• 期刊: Cell host & microbe
• 影响因子: 30.3
• 作者: S. Yamayoshi;T. Noda;H. Ebihara;H. Goto;Y. Morikawa;I. Lukashevich;G. Neumann;H. Feldmann;Y. Kawaoka