Regulation of the influenza A virus polymerase activity by the viral nuclear export protein (NEP)

病毒核输出蛋白（NEP）对甲型流感病毒聚合酶活性的调节

• 批准号: 61949751
• 负责人: Professor Dr. Martin Schwemmle
• 金额: --
• 依托单位: Institut für Medizinische Mikrobiologie und Hygiene (IMMH)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/61949751?language=en
• 关键词: Regulation; influenza; virus; polymerase; activity

Influenza A viruses cause respiratory febrile illnesses in humans, resulting in significant morbidity and mortality in the population. Although the polymerase complex of these viruses generates all viral RNA species, we and others could provide evidence that NEP is required for efficient viral polymerase activity by stimulating vRNA and cRNA synthesis, suggesting that this protein acts as a co-factor supporting viral replication. We could further show that a single adaptive mutation in the NEP of a human H5N1 isolate increased the replication efficiency of the viral polymerase and the pathogenicity of this isolate in mice. Furthermore, we obtained preliminary evidence that NEP dimer formation is required for the polymerase co-factor activity of this protein. Using recombinant protein expression, biochemical characterization, reverse genetic approaches and in-vitro transcription/replication assays, we want to gain insights into the mechanisms by which NEP increases the activity of the viral polymerase and prove our hypothesis that NEP is a component of a viral „replicase“ complex. This project will not only provide new insights into the function of NEP in viral replication but also provide the basis for future antiviral interventions.

甲型流感病毒会引起人类呼吸道发热性疾病，导致人群发病率和死亡率显着提高。尽管这些病毒的聚合酶复合物产生所有病毒RNA种类，但我们和其他人可以提供证据，证明NEP是通过刺激vRNA和cRNA合成来实现有效病毒聚合酶活性所必需的，这表明该蛋白质充当支持病毒复制的辅助因子。我们可以进一步证明，人类 H5N1 分离株 NEP 中的单个适应性突变提高了病毒聚合酶的复制效率以及该分离株对小鼠的致病性。此外，我们获得了初步证据表明 NEP 二聚体的形成是该蛋白质的聚合酶辅因子活性所必需的。利用重组蛋白表达、生化表征、反向遗传方法和体外转录/复制测定，我们希望深入了解 NEP 增加病毒聚合酶活性的机制，并证明我们的假设，即 NEP 是病毒“复制酶”复合物的一个组成部分。该项目不仅将为NEP在病毒复制中的功能提供新的见解，而且为未来的抗病毒干预措施提供基础。

项目成果

Adaptive Mutations in the Nuclear Export Protein of Human-Derived H5N1 Strains Facilitate a Polymerase Activity-Enhancing Conformation

• DOI: 10.1128/jvi.01495-13
• 发表时间: 2013-10
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Peter Reuther;S. Giese;Veronika Götz;Normann Kilb;B. Mänz;Linda Brunotte;M. Schwemmle

Phosphorylation of Highly Conserved Serine Residues in the Influenza A Virus Nuclear Export Protein NEP Plays a Minor Role in Viral Growth in Human Cells and Mice

• DOI: 10.1128/jvi.00854-14
• 发表时间: 2014-07-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Reuther, Peter;Giese, Sebastian;Schwemmle, Martin
• 通讯作者: Schwemmle, Martin

Adaptive mutations in NEP compensate for defective H5N1 RNA replication in cultured human cells

• DOI: 10.1038/ncomms1804
• 发表时间: 2012-05-01
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Maenz, Benjamin;Brunotte, Linda;Schwemmle, Martin
• 通讯作者: Schwemmle, Martin