新的犬ANP32A的克隆及其在流感病毒跨物种感染中的作用

doi:10.11843/j.issn.0366-6964.2024.02.023

Abstract

Abstract: The species-specific acidic nuclear phosphoprotein 32A (ANP32A) regulates RNA polymerase activity of influenza A virus from different hosts. To analyze species-specific difference of canine ANP32A (canine ANP32A, caANP32A) and its role in the cross-species infection of influenza virus, RT-PCR was used to amplify and clone caANP32A from Madin-Darby canine kidney (MDCK) cells and different tissues of canine lung, spleen and intestine. The interaction of caANP32A with influenza A virus RNA polymerase was analyzed by Laser confocal assay. The effect of caANP32A on RNA polymerase activity of influenza A virus was detected in 293T cells by double luciferase reporter gene experiment. The results showed that the new caANP32A was amplified from MDCK cells, which had four more amino acids insertion than the previously reported caANP32A. The new caANP32A was amplified from the lung, spleen and intestine of dogs; After sequencing and analyzing the ANP32A gene, it was found that the new caANP32A was not formed by alternative splicing of mRNA; Laser confocal assay showed that the co-location of the new caANP32A with the RNA polymerase of H3N2 CIV in the nucleus. Polymerase activity experiments showed that overexpression of novel caANP32A in mammalian cells did not promote the RNA polymerase activity of H9N2 avian influenza virus (AIV) and H3N2 canine influenza virus (CIV), but chANP32A could. The new caANP32A cloned in this study, compared to previous reports, has four amino acids LSLV insertion at positions 176 to 179, but the new caANP32A still has species restriction on the polymerase activity of AIV and does not enhance the RNA polymerase activity of CIV. This study provides a basis for further elucidating the role of dogs in the cross-species infection of influenza virus.

Key words: influenza A virus (IAV), canine acidic nuclear phosphoprotein 32 (caANP32A), RNA polymerase activity, mammalian adaptability

CLC Number:

S852.659.5

BI Zhenwei, WANG Wenjie, LIU Yakun, PENG Daxin. Cloning of a New Canine ANP32A and Its Role in Cross-species Infection of Influenza Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 660-669.

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Cloning of a New Canine ANP32A and Its Role in Cross-species Infection of Influenza Virus

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