Replication of influenza virus

流感病毒的复制

基本信息

批准号：
7924259
负责人：
Robert M Krug
金额：
$ 8.56万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-26 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7924259
关键词：
Accounting Amino Acids Animal Model Antiviral Agents Antiviral Response Applications Grants Attenuated Attenuated Live Virus Vaccine Benchmarking Binding Binding Sites Biochemical Biological Assay Birds Cell physiology Cells Cercopithecine Herpesvirus 1 Complex Defect Development Double-Stranded RNA Enzymes Event Future Generations Genes Genetic Goals Grant Hemagglutinin Human Human Influenza A Virus Inactivated Vaccines Infection Influenza Influenza A Virus, H1N1 Subtype Influenza A Virus, H3N2 Subtype Influenza A Virus, H5N1 Subtype Influenza A virus Influenza B virus Interferon Activation Interferons Laboratories Lead Life Ligase Lung diseases Mediating Messenger RNA Molecular Mus N-terminal National Institute of Allergy and Infectious Disease Neuraminidase Nuclear Magnetic Resonance Oligonucleotides PTPN11 gene Pathway interactions Play Polymerase Population Principal Investigator Process Production Protein Binding Proteins RNA Binding RNA Recognition Motif RNA chemical synthesis Reagent Recombinants Regulation Research Ribonucleases Roentgen Rays Role Simian B disease Site Structure Surface Testing Transfection Ubiquitin Vaccines Vero Cells Viral Viral Physiology Viral Proteins Virulence Virulence Factors Virus Virus Diseases Virus Replication Work attenuation base cellular targeting human G1P2 protein immunogenic influenzavirus insight mRNA Precursor mortality mutant pandemic disease prevent programs recombinant virus research study vaccine-induced immunity viral RNA

项目摘要

DESCRIPTION (provided by applicant): Influenza A and B viruses cause a highly contagious respiratory disease in humans. Influenza B viruses infect only humans, whereas influenza A viruses infect many avian and mammalian species. Influenza A viruses are responsible for the periodic wide-spread pandemics that result in high mortality rates. The 1918 pandemic was the most devastating. Highly pathogenic H5N1 influenza A viruses are prime candidates for causing the next pandemic. The NS1 proteins of influenza A virus (NS1A protein) and of influenza B virus (NS1B protein) are multifunctional proteins that suppress cellular antiviral responses and are virulence factors. Many of the functions of the NS1A protein differ from those of the NS1B protein. The overall aim is to elucidate how multiple functions of the NS1A and NS1B proteins play important roles during infection. The NS1A protein has a binding site for double-stranded RNA (dsRNA) and binding sites for several cellular proteins, including CPSF30, a cellular factor required for the processing of cellular pre-mRNAs. CPSF30 binding, which results in the inhibition of the production of all cellular mRNAs, including interferon-2 (IFN-2) mRNA, is the primary, if not the only, mechanism by which the NS1A protein of a human influenza A virus (isolated in 1972) inhibits IFN-2 mRNA production. One aim is to determine whether this mechanism is shared by all other human influenza A viruses, including H5N1 and 1918 viruses. This research will entail the generation of multiple recombinant viruses expressing NS1A proteins containing specific amino acid changes. Recent structural results on the dsRNA- and CPSF30-binding sites of the NS1A protein described in this grant application may lead to the development of new antivirals directed at these binding sites and also to the development of better live attenuated virus vaccines against H5N1 viruses. For the latter purpose, one aim is to generate viruses whose attenuation is fine-tuned by site-specific amino acid changes in these two NS1A binding sites. Motivated by genetic experiments that showed an unexpected functional interaction between the NS1A protein and the viral polymerase, another aim is to use biochemical approaches to elucidate the molecular mechanisms underlying this interaction. The IFN-induced ISG15 protein, which is conjugated to multiple cellular proteins, has antiviral activity against both influenza A and B viruses. One aim is to identify the viral and/or cellular proteins whose conjugation to ISG15 accounts for the antiviral action of ISG15 conjugation against influenza A virus. Because the NS1B protein, unlike the NS1A protein, binds ISG15 and prevents its conjugation to target proteins, research on the antiviral function of ISG15 conjugation against influenza B virus will analyze the defects in replication of a recombinant influenza B virus that expresses a NS1B protein lacking an ISG15 binding site. As an integral part of this research, the role of the dsRNA-binding activity and other functions of the NS1B protein will also be determined. Elucidation of the molecular mechanisms by which ISG15 conjugation inhibits influenza A and B viruses may lead to therapies that exploit these mechanisms.

描述（由申请人提供）：甲型和乙型流感病毒在人类中引起高度传染性呼吸道疾病。乙型流感病毒仅感染人类，而甲型流感病毒则感染许多鸟类和哺乳动物。甲型流感病毒造成周期性大范围流行，导致高死亡率。 1918 年的大流行是最具破坏性的。高致病性 H5N1 甲型流感病毒是引发下一次大流行的主要候选病毒。甲型流感病毒（NS1A 蛋白）和乙型流感病毒（NS1B 蛋白）的 NS1 蛋白是抑制细胞抗病毒反应的多功能蛋白，也是毒力因子。 NS1A 蛋白的许多功能与 NS1B 蛋白的功能不同。总体目标是阐明 NS1A 和 NS1B 蛋白的多种功能如何在感染过程中发挥重要作用。 NS1A 蛋白具有双链 RNA (dsRNA) 的结合位点和多种细胞蛋白的结合位点，其中包括 CPSF30（一种处理细胞前 mRNA 所需的细胞因子）。 CPSF30 结合会抑制所有细胞 mRNA 的产生，包括干扰素 2 (IFN-2) mRNA，这是人甲型流感病毒 NS1A 蛋白（分离的）的主要（如果不是唯一）机制。 1972 年）抑制 IFN-2 mRNA 的产生。一个目的是确定所有其他人类甲型流感病毒（包括 H5N1 和 1918 病毒）是否都具有这种机制。这项研究将需要产生多种表达含有特定氨基酸变化的 NS1A 蛋白的重组病毒。本拨款申请中描述的 NS1A 蛋白的 dsRNA 和 CPSF30 结合位点的最新结构结果可能会导致针对这些结合位点的新型抗病毒药物的开发，以及针对 H5N1 病毒的更好的减毒活病毒疫苗的开发。对于后一个目的，一个目标是产生通过这两个 NS1A 结合位点的位点特异性氨基酸变化来微调其减毒效果的病毒。遗传实验显示 NS1A 蛋白和病毒聚合酶之间存在意想不到的功能相互作用，受此启发，另一个目标是使用生化方法来阐明这种相互作用背后的分子机制。 IFN 诱导的 ISG15 蛋白与多种细胞蛋白缀合，对甲型和乙型流感病毒均具有抗病毒活性。一个目的是鉴定与 ISG15 缀合的病毒和/或细胞蛋白，解释 ISG15 缀合对甲型流感病毒的抗病毒作用。由于NS1B蛋白与NS1A蛋白不同，它结合ISG15并阻止其与靶蛋白缀合，因此ISG15缀合对乙型流感病毒的抗病毒功能的研究将分析表达缺乏NS1B蛋白的重组乙型流感病毒的复制缺陷。 ISG15 结合位点。作为本研究的一个组成部分，还将确定 NS1B 蛋白的 dsRNA 结合活性和其他功能的作用。阐明 ISG15 结合抑制甲型和乙型流感病毒的分子机制可能会产生利用这些机制的治疗方法。