Differential mitochondrial proteomic analysis of A549 cells infected with avian influenza virus subtypes H5 and H9
Abstract Background: Both the highly pathogenic avian influenza (HPAI) H5N1 and low pathogenic avian influenza (LPAI) H9N2 viruses have been reported to cross species barriers to infect humans. However, H5N1 viruses can cause severe damage and a high mortality rate, but H9N2 viruses can not. Our purpose was to use proteomics technology to study the differential expression of mitochondrial-related proteins caused by H5N1 and H9N2 virus infections.Methods: According to the determined viral infection titer, A549 cells were infected with 1 MOI (multiplicity of infection) virus, and the mitochondria were extracted after 24 hours of incubation. The lysed mitochondrial protein was analyzed by BCA method for protein concentration, SDS-PAGE preliminary analysis, two-dimensional gel electrophoresis, and mass spectrometry. Select different protein spots, perform Western Blot to verify the proteomics results. The identified proteins use GO analysis for subcellular localization, KEGG analysis for functional classification and signal pathways, STRING analysis for functional protein association networks.Results: In the 2-D gel electrophoresis analysis, 227 protein spots were detected in the H5N1-infected group, and 169 protein spots were detected in the H9N2-infected group. After further MS identification and removal of redundancy, 32 differentially expressed proteins were identified. Compared with the H9N2 group, the H5N1-infected group had 16 upregulated mitochondrial proteins and 16 downregulated proteins. The 70 kDa heat shock protein analogs, short-chain enoyl-CoA hydratase, malate dehydrogenase, and ATP synthase were verified by Western Blot, and the results were consistent with proteomics. Functional analysis indicated that these differentially expressed proteins were involved mainly in apoptosis, metabolism.Conclusions: Compared with H9N2 group, the differential expression of eight mitochondrial proteins in H5N1 group led to T cell activation, antigen presentation, stress response, ATP synthesis and cell apoptosis reduction, leading to higher pathogenicity of H5N1 than H9N2 virus..
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Preprint| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
ResearchSquare.com - (2022) vom: 29. Juli Zur Gesamtaufnahme - year:2022 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Yang, Yuting [VerfasserIn] |
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| doi: |
10.21203/rs.3.rs-60533/v2 |
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| PPN (Katalog-ID): |
XRA034380760 |
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| 245 | 1 | 0 | |a Differential mitochondrial proteomic analysis of A549 cells infected with avian influenza virus subtypes H5 and H9 |
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| 520 | |a Abstract Background: Both the highly pathogenic avian influenza (HPAI) H5N1 and low pathogenic avian influenza (LPAI) H9N2 viruses have been reported to cross species barriers to infect humans. However, H5N1 viruses can cause severe damage and a high mortality rate, but H9N2 viruses can not. Our purpose was to use proteomics technology to study the differential expression of mitochondrial-related proteins caused by H5N1 and H9N2 virus infections.Methods: According to the determined viral infection titer, A549 cells were infected with 1 MOI (multiplicity of infection) virus, and the mitochondria were extracted after 24 hours of incubation. The lysed mitochondrial protein was analyzed by BCA method for protein concentration, SDS-PAGE preliminary analysis, two-dimensional gel electrophoresis, and mass spectrometry. Select different protein spots, perform Western Blot to verify the proteomics results. The identified proteins use GO analysis for subcellular localization, KEGG analysis for functional classification and signal pathways, STRING analysis for functional protein association networks.Results: In the 2-D gel electrophoresis analysis, 227 protein spots were detected in the H5N1-infected group, and 169 protein spots were detected in the H9N2-infected group. After further MS identification and removal of redundancy, 32 differentially expressed proteins were identified. Compared with the H9N2 group, the H5N1-infected group had 16 upregulated mitochondrial proteins and 16 downregulated proteins. The 70 kDa heat shock protein analogs, short-chain enoyl-CoA hydratase, malate dehydrogenase, and ATP synthase were verified by Western Blot, and the results were consistent with proteomics. Functional analysis indicated that these differentially expressed proteins were involved mainly in apoptosis, metabolism.Conclusions: Compared with H9N2 group, the differential expression of eight mitochondrial proteins in H5N1 group led to T cell activation, antigen presentation, stress response, ATP synthesis and cell apoptosis reduction, leading to higher pathogenicity of H5N1 than H9N2 virus. | ||
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| 700 | 1 | |a Yang, Changcheng |e verfasserin |4 aut | |
| 700 | 1 | |a Fang, Fang |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Ying |e verfasserin |4 aut | |
| 700 | 1 | |a Chang, Haiyan |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Ze |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Ping |e verfasserin |4 aut | |
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