S-Acylation of hemmagglutinin of influenza virus with different fatty acids - structural requirements and functional consequences

不同脂肪酸对流感病毒血凝素的 S-酰化 - 结构要求和功能后果

• 批准号: 125542159
• 负责人: Privatdozent Dr. Michael Veit
• 金额: --
• 依托单位: Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/125542159?language=en
• 关键词: Acylation; hemmagglutinin; influenza; virus; different

The hemagglutinin (HA) of influenza virus is S-acylated at three cysteines located either in the inner leaflet of the transmembrane region (TMR) or in the cytoplasmic tail (CT). Acylation of HA is essential for virus replication, affecting cell entry of viruses via membrane fusion and/or budding of virus particles from the infected cell. In a previous collaboration between Berlin (Veit) and Moscow (Kordyukova) we used mass-spectrometry and MS/MS sequencing to show that the cysteine located in the TMR of HA contains only stearate whereas two cysteines in the CT are modified with palmitate. With this joint project proposal we attempt to identify the structural basis for differential acylation and explore the significance of the type of fatty acids for the function of HA. Prof. Efremov, a specialist for computer modelling (molecular dynamics and Monte Carlo simulations), will join the project to determine the conformation of the TMR and CT of HA (and of other viral acylproteins) to identify peculiar signals present around palmitoylated and stearoylated cysteine residues. The predictions will be tested by mutagenesis, expression and fatty acid analysis of HA. Finally, using three already available recombinant influenza viruses, each having HA with one fatty acid binding site deleted (and as a consequence large differences in their stearate content), we will analyze whether S-acylation affects binding of HA to the matrixprotein M1, which is a prerequisite for the budding of infectious virus particles.

流感病毒的血凝素(HA)由位于跨膜区(TMR)内叶或胞浆尾部(CT)的三个半胱氨酸S酰化而成。HA的酰化是病毒复制的关键，它通过膜融合和/或感染细胞中的病毒颗粒萌发来影响病毒进入细胞。在柏林(Veit)和莫斯科(Kordyukova)之前的一次合作中，我们使用质谱仪和MS/MS测序表明，位于HA的TMR中的半胱氨酸只含有硬脂酸盐，而CT中的两个半胱氨酸被棕榈酸酯修饰。通过这个联合项目提案，我们试图确定差异酰化的结构基础，并探索脂肪酸类型对HA功能的意义。计算机模拟(分子动力学和蒙特卡罗模拟)专家Efemov教授将加入这个项目，以确定HA(和其他病毒酰基蛋白)的TMR和CT的构象，以确定棕榈酰化和硬脂酰化半胱氨酸残基周围存在的特殊信号。这些预测将通过对HA的突变、表达和脂肪酸分析来验证。最后，使用三种已有的重组流感病毒，每种病毒的HA都有一个脂肪酸结合位点缺失(因此它们的硬脂酸含量存在很大差异)，我们将分析S酰化是否影响HA与基质蛋白M1的结合，这是传染性病毒颗粒萌发的先决条件。