Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein
Abstract Background Influenza is one of the most important viral infections globally. Viral RNA-dependent RNA polymerase (RdRp) consists of the PA, PB1, and PB2 subunits, and the amino acid residues of each subunit are highly conserved among influenza A virus (IAV) strains. Due to the high mutation...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-02-01
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| Series: | Journal of Biomedical Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12929-023-00901-x |
| _version_ | 1797863953243570176 |
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| author | Mei-Lin Yang Yi-Cheng Chen Chung-Teng Wang Hao-Earn Chong Nai-Hui Chung Chia-Hsing Leu Fu-Tong Liu Michael M. C. Lai Pin Ling Chao-Liang Wu Ai-Li Shiau |
| author_facet | Mei-Lin Yang Yi-Cheng Chen Chung-Teng Wang Hao-Earn Chong Nai-Hui Chung Chia-Hsing Leu Fu-Tong Liu Michael M. C. Lai Pin Ling Chao-Liang Wu Ai-Li Shiau |
| author_sort | Mei-Lin Yang |
| collection | DOAJ |
| description | Abstract Background Influenza is one of the most important viral infections globally. Viral RNA-dependent RNA polymerase (RdRp) consists of the PA, PB1, and PB2 subunits, and the amino acid residues of each subunit are highly conserved among influenza A virus (IAV) strains. Due to the high mutation rate and emergence of drug resistance, new antiviral strategies are needed. Host cell factors are involved in the transcription and replication of influenza virus. Here, we investigated the role of galectin-3, a member of the β-galactoside-binding animal lectin family, in the life cycle of IAV infection in vitro and in mice. Methods We used galectin-3 knockout and wild-type mice and cells to study the intracellular role of galectin-3 in influenza pathogenesis. Body weight and survival time of IAV-infected mice were analyzed, and viral production in mouse macrophages and lung fibroblasts was examined. Overexpression and knockdown of galectin-3 in A549 human lung epithelial cells were exploited to assess viral entry, viral ribonucleoprotein (vRNP) import/export, transcription, replication, virion production, as well as interactions between galectin-3 and viral proteins by immunoblotting, immunofluorescence, co-immunoprecipitation, RT-qPCR, minireplicon, and plaque assays. We also employed recombinant galectin-3 proteins to identify specific step(s) of the viral life cycle that was affected by exogenously added galectin-3 in A549 cells. Results Galectin-3 levels were increased in the bronchoalveolar lavage fluid and lungs of IAV-infected mice. There was a positive correlation between galectin-3 levels and viral loads. Notably, galectin-3 knockout mice were resistant to IAV infection. Knockdown of galectin-3 significantly reduced the production of viral proteins and virions in A549 cells. While intracellular galectin-3 did not affect viral entry, it increased vRNP nuclear import, RdRp activity, and viral transcription and replication, which were associated with the interaction of galectin-3 with viral PA subunit. Galectin-3 enhanced the interaction between viral PA and PB1 proteins. Moreover, exogenously added recombinant galectin-3 proteins also enhanced viral adsorption and promoted IAV infection in A549 cells. Conclusion We demonstrate that galectin-3 enhances viral infection through increases in vRNP nuclear import and RdRp activity, thereby facilitating viral transcription and replication. Our findings also identify galectin-3 as a potential therapeutic target for influenza. |
| first_indexed | 2024-04-09T22:43:53Z |
| format | Article |
| id | doaj.art-8201530a06c646baba7c06ffdd648f2d |
| institution | Directory Open Access Journal |
| issn | 1423-0127 |
| language | English |
| last_indexed | 2024-04-09T22:43:53Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biomedical Science |
| spelling | doaj.art-8201530a06c646baba7c06ffdd648f2d2023-03-22T11:59:16ZengBMCJournal of Biomedical Science1423-01272023-02-0130112210.1186/s12929-023-00901-xUpregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA proteinMei-Lin Yang0Yi-Cheng Chen1Chung-Teng Wang2Hao-Earn Chong3Nai-Hui Chung4Chia-Hsing Leu5Fu-Tong Liu6Michael M. C. Lai7Pin Ling8Chao-Liang Wu9Ai-Li Shiau10Department of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityDepartment of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung UniversityDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityDepartment of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung UniversityDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityDepartment of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung UniversityInstitute of Biomedical Sciences, Academia SinicaGraduate Institute of Biomedical Sciences, China Medical UniversityDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityDitmanson Medical Foundation Chia-Yi Christian HospitalDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityAbstract Background Influenza is one of the most important viral infections globally. Viral RNA-dependent RNA polymerase (RdRp) consists of the PA, PB1, and PB2 subunits, and the amino acid residues of each subunit are highly conserved among influenza A virus (IAV) strains. Due to the high mutation rate and emergence of drug resistance, new antiviral strategies are needed. Host cell factors are involved in the transcription and replication of influenza virus. Here, we investigated the role of galectin-3, a member of the β-galactoside-binding animal lectin family, in the life cycle of IAV infection in vitro and in mice. Methods We used galectin-3 knockout and wild-type mice and cells to study the intracellular role of galectin-3 in influenza pathogenesis. Body weight and survival time of IAV-infected mice were analyzed, and viral production in mouse macrophages and lung fibroblasts was examined. Overexpression and knockdown of galectin-3 in A549 human lung epithelial cells were exploited to assess viral entry, viral ribonucleoprotein (vRNP) import/export, transcription, replication, virion production, as well as interactions between galectin-3 and viral proteins by immunoblotting, immunofluorescence, co-immunoprecipitation, RT-qPCR, minireplicon, and plaque assays. We also employed recombinant galectin-3 proteins to identify specific step(s) of the viral life cycle that was affected by exogenously added galectin-3 in A549 cells. Results Galectin-3 levels were increased in the bronchoalveolar lavage fluid and lungs of IAV-infected mice. There was a positive correlation between galectin-3 levels and viral loads. Notably, galectin-3 knockout mice were resistant to IAV infection. Knockdown of galectin-3 significantly reduced the production of viral proteins and virions in A549 cells. While intracellular galectin-3 did not affect viral entry, it increased vRNP nuclear import, RdRp activity, and viral transcription and replication, which were associated with the interaction of galectin-3 with viral PA subunit. Galectin-3 enhanced the interaction between viral PA and PB1 proteins. Moreover, exogenously added recombinant galectin-3 proteins also enhanced viral adsorption and promoted IAV infection in A549 cells. Conclusion We demonstrate that galectin-3 enhances viral infection through increases in vRNP nuclear import and RdRp activity, thereby facilitating viral transcription and replication. Our findings also identify galectin-3 as a potential therapeutic target for influenza.https://doi.org/10.1186/s12929-023-00901-xGalectin-3Influenza virusvRNP importRNA-dependent RNA polymeraseRNA synthesisViral PA |
| spellingShingle | Mei-Lin Yang Yi-Cheng Chen Chung-Teng Wang Hao-Earn Chong Nai-Hui Chung Chia-Hsing Leu Fu-Tong Liu Michael M. C. Lai Pin Ling Chao-Liang Wu Ai-Li Shiau Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein Journal of Biomedical Science Galectin-3 Influenza virus vRNP import RNA-dependent RNA polymerase RNA synthesis Viral PA |
| title | Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein |
| title_full | Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein |
| title_fullStr | Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein |
| title_full_unstemmed | Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein |
| title_short | Upregulation of galectin-3 in influenza A virus infection promotes viral RNA synthesis through its association with viral PA protein |
| title_sort | upregulation of galectin 3 in influenza a virus infection promotes viral rna synthesis through its association with viral pa protein |
| topic | Galectin-3 Influenza virus vRNP import RNA-dependent RNA polymerase RNA synthesis Viral PA |
| url | https://doi.org/10.1186/s12929-023-00901-x |
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