犬ANP32蛋白多态性及其对A型流感病毒RNA聚合酶活性的影响

doi:10.16431/j.cnki.1671-7236.2022.07.004

Abstract

Abstract: 【Objective】 This study was aimed to explore the effect of canine acidic (leucine-rich) nuclear phosphoprotein 32 ku (caANP32) on the RNA polymerase activity of Influenza A virus (IAV) from different species.【Method】 Real-time PCR method was used to analyze the tissue distribution of caANP32 family genes (caANP32A,caANP32B and caANP32E),then these genes were amplified and cloned to evaluate the supporting effect of caANP32 family proteins on the RNA polymerase activity of IAV in different species.【Result】 The results showed that the caANP32 family genes were similarly expressed in different tissues,and the expression abundance of caANP32B gene was significantly or extremely significantly higher than that of caANP32A and caANP32E genes in different tissues (P＜0.05;P＜0.01),the tissue abundance of caANP32E gene was extremely significantly or significantly higher than that of caANP32A gene in heart,cecum and brain (P＜0.01;P＜0.05),there was no significant difference in gene abundance between caANP32E and caANP32A genes in liver,lung and other tissues (P>0.05).Only one transcript of caANP32A gene was found in canine heart tissue and MDCK cells,however three different splice variants of caANP32B gene (caANP32B_X1,caANP32B_X2 and caANP32B_X3), and two different splice variants of the caANP32E gene (caANP32E_X1 and caANP32E_X2) were found in these samples.By analyzing the effects of ANP32 family members on the RNA polymerase activity of IAV from different species,it was found that caANP32A and caANP32B could support Canine influenza virus (H3N2_GD11) and Equine influenza virus (H3N8_JL89) RNA polymerase activity,but had lower activity to support Avian influenza virus (H9N2_ZJ12) RNA polymerase activity.The supporting ability of caANP32B_X2 to the RNA polymerase activity of IAV was significantly higher than that of caANP32B_X1 and caANP32B_X3,while caANP32E did not support the RNA polymerase activity of Avian influenza virus.【Conclusion】 This study analyzed the tissue distribution and sequence polymorphisms of the caANP32 family members,and clarified their role in supporting RNA polymerase activity of IAV from different species,providing a new basis for analyzing the molecular mechanism of MDCK as a cell line for influenza virus isolation and vaccine production.

Key words: Influenza A virus (IAV); caANP32; MDCK cells; polymerase activity

CLC Number:

S852.65

ZHANG Yuan, YU Mengmeng, SUN Liuke, GUO Xing, NA Lei, LIU Diqiu, JIANG Qian, ZHANG Haili, WANG Xiaojun. Analysis of Canine ANP32 Protein Polymorphism and Effect on RNA Polymerase Activity of Influenza A Virus[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2462-2473.

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Analysis of Canine ANP32 Protein Polymorphism and Effect on RNA Polymerase Activity of Influenza A Virus

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