Influenza PB1-F2 Inhibits Avian MAVS Signaling
RIG-I plays an essential role in the duck innate immune response to influenza infection. RIG-I engages the critical adaptor protein mitochondrial antiviral signaling (MAVS) to activate the downstream signaling pathway. The influenza A virus non-structural protein PB1-F2 interacts with MAVS in human...
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| Format: | Article |
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MDPI AG
2020-04-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/4/409 |
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| author | Yanna Xiao Danyel Evseev Chase A. Stevens Adam Moghrabi Domingo Miranzo-Navarro Ximena Fleming-Canepa David G. Tetrault Katharine E. Magor |
| author_facet | Yanna Xiao Danyel Evseev Chase A. Stevens Adam Moghrabi Domingo Miranzo-Navarro Ximena Fleming-Canepa David G. Tetrault Katharine E. Magor |
| author_sort | Yanna Xiao |
| collection | DOAJ |
| description | RIG-I plays an essential role in the duck innate immune response to influenza infection. RIG-I engages the critical adaptor protein mitochondrial antiviral signaling (MAVS) to activate the downstream signaling pathway. The influenza A virus non-structural protein PB1-F2 interacts with MAVS in human cells to inhibit interferon production. As duck and human MAVS share only 28% amino acid similarity, it is not known whether the influenza virus can similarly inhibit MAVS signaling in avian cells. Using confocal microscopy we show that MAVS and the constitutively active N-terminal end of duck RIG-I (2CARD) co-localize in DF-1 cells, and duck MAVS is pulled down with GST-2CARD. We establish that either GST-2CARD, or duck MAVS can initiate innate signaling in chicken cells and their co-transfection augments interferon-beta promoter activity. Demonstrating the limits of cross-species interactions, duck RIG-I 2CARD initiates MAVS signaling in chicken cells, but works poorly in human cells. The D122A mutation of human 2CARD abrogates signaling by affecting MAVS engagement, and the reciprocal A120D mutation in duck 2CARD improves signaling in human cells. We show mitochondrial localization of PB1-F2 from influenza A virus strain A/Puerto Rico/8/1934 (H1N1; PR8), and its co-localization and co-immunoprecipitation with duck MAVS. PB1-F2 inhibits interferon-beta promoter activity induced by overexpression of either duck RIG-I 2CARD, full-length duck RIG-I, or duck MAVS. Finally, we show that the effect of PB1-F2 on mitochondria abrogates TRIM25-mediated ubiquitination of RIG-I CARD in both human and avian cells, while an NS1 variant from the PR8 influenza virus strain does not. |
| first_indexed | 2024-03-10T20:36:28Z |
| format | Article |
| id | doaj.art-0f94628e43874006928d66872dd69332 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T20:36:28Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-0f94628e43874006928d66872dd693322023-11-19T20:56:48ZengMDPI AGViruses1999-49152020-04-0112440910.3390/v12040409Influenza PB1-F2 Inhibits Avian MAVS SignalingYanna Xiao0Danyel Evseev1Chase A. Stevens2Adam Moghrabi3Domingo Miranzo-Navarro4Ximena Fleming-Canepa5David G. Tetrault6Katharine E. Magor7Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaDepartment of Biological Sciences, University of Alberta, Edmonton, AB T6G 2R3, CanadaRIG-I plays an essential role in the duck innate immune response to influenza infection. RIG-I engages the critical adaptor protein mitochondrial antiviral signaling (MAVS) to activate the downstream signaling pathway. The influenza A virus non-structural protein PB1-F2 interacts with MAVS in human cells to inhibit interferon production. As duck and human MAVS share only 28% amino acid similarity, it is not known whether the influenza virus can similarly inhibit MAVS signaling in avian cells. Using confocal microscopy we show that MAVS and the constitutively active N-terminal end of duck RIG-I (2CARD) co-localize in DF-1 cells, and duck MAVS is pulled down with GST-2CARD. We establish that either GST-2CARD, or duck MAVS can initiate innate signaling in chicken cells and their co-transfection augments interferon-beta promoter activity. Demonstrating the limits of cross-species interactions, duck RIG-I 2CARD initiates MAVS signaling in chicken cells, but works poorly in human cells. The D122A mutation of human 2CARD abrogates signaling by affecting MAVS engagement, and the reciprocal A120D mutation in duck 2CARD improves signaling in human cells. We show mitochondrial localization of PB1-F2 from influenza A virus strain A/Puerto Rico/8/1934 (H1N1; PR8), and its co-localization and co-immunoprecipitation with duck MAVS. PB1-F2 inhibits interferon-beta promoter activity induced by overexpression of either duck RIG-I 2CARD, full-length duck RIG-I, or duck MAVS. Finally, we show that the effect of PB1-F2 on mitochondria abrogates TRIM25-mediated ubiquitination of RIG-I CARD in both human and avian cells, while an NS1 variant from the PR8 influenza virus strain does not.https://www.mdpi.com/1999-4915/12/4/409PB1-F2NS1type I interferonRIG-IduckMAVS |
| spellingShingle | Yanna Xiao Danyel Evseev Chase A. Stevens Adam Moghrabi Domingo Miranzo-Navarro Ximena Fleming-Canepa David G. Tetrault Katharine E. Magor Influenza PB1-F2 Inhibits Avian MAVS Signaling Viruses PB1-F2 NS1 type I interferon RIG-I duck MAVS |
| title | Influenza PB1-F2 Inhibits Avian MAVS Signaling |
| title_full | Influenza PB1-F2 Inhibits Avian MAVS Signaling |
| title_fullStr | Influenza PB1-F2 Inhibits Avian MAVS Signaling |
| title_full_unstemmed | Influenza PB1-F2 Inhibits Avian MAVS Signaling |
| title_short | Influenza PB1-F2 Inhibits Avian MAVS Signaling |
| title_sort | influenza pb1 f2 inhibits avian mavs signaling |
| topic | PB1-F2 NS1 type I interferon RIG-I duck MAVS |
| url | https://www.mdpi.com/1999-4915/12/4/409 |
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