Tyrosines in the Influenza A Virus M2 Protein Cytoplasmic Tail Are Critical for Production of Infectious Virus Particles
Tyrosines in the Influenza A Virus M2 Protein Cytoplasmic Tail Are Critical for Production of Infectious Virus Particles
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DOI：
10.1128/jvi.00853-10
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发表时间：
2010-09-01
影响因子：
5.4
通讯作者：
Pekosz, Andrew
中科院分区：
文献类型：
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作者：
Grantham, Michael L.;Stewart, Shaun M.;Pekosz, Andrew
甲型流感病毒M2蛋白质的胞质尾区是产生感染性病毒体所必需的。在本研究中，通过单丙氨酸扫描诱变鉴定了M2胞质尾区中的关键残基。使用反向遗传学和蛋白质反式互补测定，在迄今为止测序的>99%的流感病毒株中保守的位置76处的酪氨酸残基被鉴定为对于感染性病毒颗粒的形成至关重要。编码具有Y76A突变的M2的重组病毒在MDCK细胞以及原代分化的气道上皮细胞培养物中表现出复制缺陷，丝状病毒颗粒形成缺陷，以及核蛋白包装成病毒颗粒减少。这些缺陷都可以通过M2胞质尾区第71位丝氨酸突变为酪氨酸来克服，该突变是在含有Y76A突变的病毒盲传后出现的。这些数据证实并扩展了我们对M2蛋白对感染性病毒颗粒组装的重要性的理解。
The cytoplasmic tail of the influenza A virus M2 protein is required for the production of infectious virions. In this study, critical residues in the M2 cytoplasmic tail were identified by single-alanine scanning mutagenesis. The tyrosine residue at position 76, which is conserved in >99% of influenza virus strains sequenced to date, was identified as being critical for the formation of infectious virus particles using both reverse genetics and a protein trans-complementation assay. Recombinant viruses encoding M2 with the Y76A mutation demonstrated replication defects in MDCK cells as well as in primary differentiated airway epithelial cell cultures, defects in the formation of filamentous virus particles, and reduced packaging of nucleoprotein into virus particles. These defects could all be overcome by a mutation of serine to tyrosine at position 71 of the M2 cytoplasmic tail, which emerged after blind passage of viruses containing the Y76A mutation. These data confirm and extend our understanding of the significance of the M2 protein for infectious virus particle assembly.