Equine ANP32 proteins support influenza A virus RNA polymerase activity

Yuan Zhang; Xing Guo; Mengmeng Yu; Liuke Sun; Yuxing Qu; Kui Guo; Zhe Hu; Diqiu Liu; Haili Zhang; Xiaojun Wang

doi:10.1016/j.virs.2023.10.009

December 2023

. doi: 10.1016/j.virs.2023.10.009

Citation: Yuan Zhang, Xing Guo, Mengmeng Yu, Liuke Sun, Yuxing Qu, Kui Guo, Zhe Hu, Diqiu Liu, Haili Zhang, Xiaojun Wang. Equine ANP32 proteins support influenza A virus RNA polymerase activity .VIROLOGICA SINICA, 2023, 38(6) : 951-960. http://dx.doi.org/10.1016/j.virs.2023.10.009

马ANP32蛋白支持A型流感病毒RNA聚合酶活性

张媛 ^a ,
郭兴 ^a ,
于萌萌 ^a ,
孙留克 ^a ,
屈玉杏 ^a ,
郭奎 ^a ,
胡哲 ^a ,
刘荻萩 ^a ,
张海丽 ^b,, ,
王晓钧 ^a,,

a. 中国农业科学院哈尔滨兽医研究所, 动物疫病防控全国重点实验室;
b. 吉林大学动物医学学院, 人畜共患传染病重症诊治全国重点实验室, 人兽共患病研究教育部重点实验室

通讯作者： 张海丽, zhanghaili@jlu.edu.cn; 王晓钧, wangxiaojun@caas.cn
收稿日期： 2023-05-14
录用日期： 2023-10-23

摘要

宿主ANP32家族蛋白是支持流感病毒RNA聚合酶活性的关键蛋白，其在流感病毒跨物种传播中也发挥重要作用。迄今为止，马属动物ANP32（Equine ANP32，eqANP32）的分子特性及其影响马流感病毒（equine influenza virus，EIV） RNA聚合酶活性的机制尚不清楚。在本研究中，作者发现eqANP32A存在六种表达水平不同的剪接变体。进一步研究发现，eqANP32A六个剪接变体不同程度地支持EIV RNA聚合酶活性，其中eqANP32A_X2剪接变体表达丰度最高，对RNA聚合酶活性支持也最高。序列分析表明，eqANP32A六个剪接变体的N-Cap区域差异显著影响其N端构象，但不影响其结合RNA聚合酶活性的能力。此外，作者还发现eqANP32B只有一个转录本且其对EIV RNA聚合酶支持活性较弱，是源于其C末端111-259氨基酸序列所导致的功能缺陷。作者的研究表明，eqANP32A_X2蛋白是决定EIV在马匹内有效复制的关键宿主因子。综上所述，作者的研究分析了eqANP32家族蛋白的分子特性，揭示了eqANP32A和eqANP32B的序列特征，并首次提示ANP32A蛋白的N-cap区域在支持流感病毒聚合酶的活性中也发挥了重要作用。
- 马流感病毒
- , 马ANP32A
- , 马ANP32B
- , RNA聚合酶活性
- , N-Cap结构域

Equine ANP32 proteins support influenza A virus RNA polymerase activity

Yuan Zhang ^a ,
Xing Guo ^a ,
Mengmeng Yu ^a ,
Liuke Sun ^a ,
Yuxing Qu ^a ,
Kui Guo ^a ,
Zhe Hu ^a ,
Diqiu Liu ^a ,
Haili Zhang ^b,, ,
Xiaojun Wang ^a,,

a. State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin, 150069, China;
b. State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis and College of Veterinary Medicine, Jilin University, Changchun, 130062, China

Corresponding author: Haili Zhang, zhanghaili@jlu.edu.cn Xiaojun Wang, wangxiaojun@caas.cn
Received Date: 14 May 2023
Accepted Date: 23 October 2023

Abstract

Host ANP32 family proteins are crucial for maintaining the activity of influenza RNA polymerase and play an important role in the cross-species transmission of influenza viruses. To date, the molecular properties of equine ANP32 (eqANP32) protein are poorly understood, particularly the mechanisms that affect equine influenza virus (EIV) RNA polymerase activity. Here, we found that there are six alternative splicing variants of equine ANP32A (eqANP32A) with different levels of expression. Further studies showed that these six splicing variants of eqANP32A supported the activity of EIV RNA polymerase to varying degrees, with the variant eqANP32A_X2 having the highest expression abundance and exhibiting the highest support of polymerase activity. Sequence analysis demonstrated that the differences in the N-Cap regions of the six splicing variants significantly affected their N-terminal conformation, but did not affect their ability to bind RNA polymerase. We also demonstrated that there is only one transcript of eqANP32B, and that this transcript showed only very low support to the EIV RNA polymerase. This functional defect in eqANP32B is caused by the sequence of the 110–259 amino acids at its C-terminus. Our results indicated that it is the eqANP32A_X2 protein that mainly determines the efficiency of the EIV replication in horses. In conclusion, our study parsed the molecular properties of eqANP32 family proteins and revealed the sequence features of eqANP32A and eqANP32B, suggesting for the first time that the N-Cap region of ANP32A protein also plays an important role in supporting the activity of the influenza virus polymerase.
- Equine influenza virus (EIV)
- , Equine ANP32A
- , Equine ANP32B
- , RNA polymerase activity
- , N-Cap domain

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Electronic Supplementary Material

10.1016j.virs.2023.10.009-ESM.docx