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Journal of Integrative Agriculture  2024, Vol. 23 Issue (9): 3145-3158    DOI: 10.1016/j.jia.2023.11.008
Special Issue: 动物病毒Animal Virology
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Role of feline ANP32 proteins in regulating polymerase activity of influenza A virus

Gang Lu1, Feiyan Zheng1, Yuqing Xiao1, Ran Shao1, Jiajun Ou1, Xin Yin2#, Shoujun Li1#, Guihong Zhang1, 3, 4#

1 College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
2 State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150000, China
3 Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming 525000, China
4 Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, China

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摘要  

近年来,越来越多的自然感染病例报道及动物攻毒实验证实猫科动物可以感染多种亚型的A型流感病毒。值得注意的是,某些亚型A型流感病毒在跨物种传播后可以在猫科动物中长期流行,甚至可以感染人类。这对公共卫生安全造成了潜在威胁。与A型流感病毒聚合酶活性相互作用的宿主因子是决定其感染宿主范围的关键因素,而ANP32蛋白是其中重要的一个宿主因子。但是,猫源ANP32蛋白对A型流感病毒聚合酶活性的调控作用及其对病毒感染宿主范围的潜在影响仍属未知领域。在本研究中,利用RT-PCR方法从家猫组织中共克隆了10个猫源ANP32剪接变异体,其中分别包括5个ANP32A、3个ANP32B及2个ANP32E变异体。测序及蛋白序列比对的结果表明部分猫源ANP32剪接变异体发生了氨基酸的缺失及/或插入。激光共聚焦实验的研究结果表明所有10个猫源ANP32剪接变异体都主要定位于细胞核。在本研究中,还建立了多个A型流感病毒代表性毒株的小基因组复制系统。利用该系统,评估了猫源ANP32蛋白对A型流感病毒聚合酶活性的调控作用。研究结果表明大部分猫源ANP32A及ANP32B蛋白都可以支持所检测的A型流感病毒的聚合酶活性,但是同一种ANP32蛋白对不同A型流感病毒毒株聚合酶活性的支持作用有所差别。此外,体外研究的结果证实了猫源ANP32蛋白可以支持H3N2犬流感病毒在细胞中复制。综上所述,本研究系统分析了猫源ANP32A蛋白对A型流感病毒聚合酶活性的调控作用,为研究猫科动物对A型流感病毒易感的分子机制提供了基础。



Abstract  

Recently, increasing natural infection cases and experimental animal challenge studies demonstrated domestic cats are susceptible to multiple subtypes influenza A virus (IAV) infections.  Notably, some subtype IAV strains could circulate in domestic cats after cross-species transmission and even infected humans, posing a threat to public health.  Host factors related to viral polymerase activity could determine host range of IAV and acidic nuclear phosphoprotein 32 (ANP32) is the most important one among them.  However, role of cat-derived ANP32 on viral polymerase activity and host range of IAV is still unknown.  In the present study, a total of 10 feline ANP32 (feANP32) splice variants (including 5 feANP32A, 3 feANP32B, and 2 feANP32E) were obtained from domestic cats by RT-PCR.  Sequence alignment results demonstrated amino acid deletions and/or insertions occurred among feANP32 variants, but all feANP32 proteins were primarily localized to cell nucleus.  Minigenome replication systems for several representative IAV strains were established and the support ability of feANP32 on IAV polymerase activity was estimated.  The results indicated that most feANP32A and feANP32B splice variants were able to support all the tested IAV strains, though the support activity of a single feANP32 protein on polymerase activity varied among different IAV strains.  In addition, the role of feANP32 in supporting H3N2 canine influenza virus was determined by investigating viral replication in vitro.  Collectively, our study systematically investigated the support activity of feANP32 on IAV, providing a clue for further exploring the mechanism of susceptibility of cats to IAV.

Keywords:  ANP32       influenza A virus       feline       cross-species transmission  
Received: 05 July 2023   Accepted: 30 October 2023
Fund: 
The authors were supported by the National Natural Science Foundation of China (32172826).
About author:  #Correspondence Xin Yin, Tel: +86-451-51051745, E-mail: yinxin@caas.cn; Shoujun Li, Tel: +86-532-85280234, E-mail: shoujunli@scau.edu.cn; Guihong Zhang, Tel: +86-532-85280234, E-mail: guihongzh@scau.edu.cn

Cite this article: 

Gang Lu, Feiyan Zheng, Yuqing Xiao, Ran Shao, Jiajun Ou, Xin Yin, Shoujun Li, Guihong Zhang. 2024. Role of feline ANP32 proteins in regulating polymerase activity of influenza A virus. Journal of Integrative Agriculture, 23(9): 3145-3158.

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