Naturally occurring PA<sup>E206K</sup> point mutation in 2009 H1N1 pandemic influenza viruses impairs viral replication at high temperatures

Mengmeng Cao; Qiannan Jia; Jinghua Li; Lili Zhao; Li zhu; Yufan Zhang; Shan Li; Tao Deng

doi:10.1016/j.virs.2023.11.005

February 2024

. doi: 10.1016/j.virs.2023.11.005

Citation: Mengmeng Cao, Qiannan Jia, Jinghua Li, Lili Zhao, Li zhu, Yufan Zhang, Shan Li, Tao Deng. Naturally occurring PA^E206K point mutation in 2009 H1N1 pandemic influenza viruses impairs viral replication at high temperatures .VIROLOGICA SINICA, 2024, 39(1) : 71-80. http://dx.doi.org/10.1016/j.virs.2023.11.005

H1N1流感病毒中自然发生的PA E206K点突变在高温下不利于流感病毒的复制

曹萌萌 ^a ,
贾倩楠 ^a ,
李静华 ^a ,
赵丽丽 ^a ,
朱莉 ^b ,
张宇凡 ^b ,
李珊 ^b ,
邓涛 ^a,b,,

a. 中国医学科学院/北京协和医学院病原生物学研究所;
b. 中国科学院微生物研究所

通讯作者： 邓涛, dengt@im.ac.cn
收稿日期： 2023-07-18
录用日期： 2023-11-10

摘要

2009年4月，甲型H1N1流感病毒的出现标志着21世纪疾病的第一次大流行。在这项研究中，我们观察到从中国和日本患者分离到的两个2009甲型H1N1流感病毒株的聚合酶活性存在明显的差异。病毒RNA依赖性RNA聚合酶复合物的三个主要蛋白质亚基（PB2、PB1和PA）的序列比对和随后的突变分析表明，PA蛋白的单个氨基酸的突变（E206K）是影响流感病毒复制发生改变的原因。进一步的实验表明，PAE206K的存在降低了A/WSN/33在哺乳动物细胞中的复制能力，同时也降低了流感病毒在体内的致病性。而分子机制研究表明，PA E206K是一种温度敏感的突变，在高温（39.5℃）下无法将PB1-PA复合物转运到细胞核。因此，PA蛋白中这种天然存在的变体是开发减毒流感活疫苗的候选突变。
- 流感病毒
- , PA蛋白
- , 点突变
- , 病毒复制

Naturally occurring PA^E206K point mutation in 2009 H1N1 pandemic influenza viruses impairs viral replication at high temperatures

Mengmeng Cao ^a ,
Qiannan Jia ^a ,
Jinghua Li ^a ,
Lili Zhao ^a ,
Li zhu ^b ,
Yufan Zhang ^b ,
Shan Li ^b ,
Tao Deng ^a,b,,

a. National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100730, China;
b. CAS Key Laboratory of Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China

Corresponding author: Tao Deng, dengt@im.ac.cn
Received Date: 18 July 2023
Accepted Date: 10 November 2023

Abstract

The emergence of influenza virus A pandemic H1N1 in April 2009 marked the first pandemic of the 21st century. In this study, we observed significant differences in the polymerase activities of two clinical 2009 H1N1 influenza A virus isolates from Chinese and Japanese patients. Sequence comparison of the three main protein subunits (PB2, PB1, and PA) of the viral RNA-dependent RNA polymerase complex and subsequent mutational analysis revealed that a single amino acid substitution (E206K) was responsible for the observed impaired replication phenotype. Further in vitro experiments showed that presence of PA^E206K decreased the replication of influenza A/WSN/33 virus in mammalian cells and a reduction in the virus’s pathogenicity in vivo. Mechanistic studies revealed that PA^E206K is a temperature-sensitive mutant associated with the inability to transport PB1–PA complex to the nucleus at high temperature (39.5 ℃). Hence, this naturally occurring variant in the PA protein represents an ideal candidate mutation for the development of live attenuated influenza vaccines.
- H1N1
- , Influenza A virus
- , Polymerase acidic protein
- , Point mutation
- , Viral replication

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