Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane
The fully assembled influenza A virus (IAV) has on its surface the highest density of a single membrane protein found in nature—the glycoprotein hemagglutinin (HA) that mediates viral binding, entry, and assembly. HA clusters at the plasma membrane of infected cells, and the HA density (number of mo...
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MDPI AG
2022-11-01
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| Online Access: | https://www.mdpi.com/1999-4915/14/11/2509 |
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| author | Prakash Raut Bright Obeng Hang Waters Joshua Zimmerberg Julie A. Gosse Samuel T. Hess |
| author_facet | Prakash Raut Bright Obeng Hang Waters Joshua Zimmerberg Julie A. Gosse Samuel T. Hess |
| author_sort | Prakash Raut |
| collection | DOAJ |
| description | The fully assembled influenza A virus (IAV) has on its surface the highest density of a single membrane protein found in nature—the glycoprotein hemagglutinin (HA) that mediates viral binding, entry, and assembly. HA clusters at the plasma membrane of infected cells, and the HA density (number of molecules per unit area) of these clusters correlates with the infectivity of the virus. Dense HA clusters are considered to mark the assembly site and ultimately lead to the budding of infectious IAV. The mechanism of spontaneous HA clustering, which occurs with or without other viral components, has not been elucidated. Using super-resolution fluorescence photoactivation localization microscopy (FPALM), we have previously shown that these HA clusters are interdependent on phosphatidylinositol 4,5-biphosphate (PIP2). Here, we show that the IAV matrix protein M1 co-clusters with PIP2, visualized using the pleckstrin homology domain. We find that cetylpyridinium chloride (CPC), which is a positively charged quaternary ammonium compound known for its antibacterial and antiviral properties at millimolar concentrations, disrupts M1 clustering and M1-PIP2 co-clustering at micromolar concentrations well below the critical micelle concentration (CMC). CPC also disrupts the co-clustering of M1 with HA at the plasma membrane, suggesting the role of host cell PIP2 clusters as scaffolds for gathering and concentrating M1 and HA to achieve their unusually high cluster densities in the IAV envelope. |
| first_indexed | 2024-03-09T17:56:14Z |
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| id | doaj.art-71b50dbe7d494af092ec0e1e3b64f3b3 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T17:56:14Z |
| publishDate | 2022-11-01 |
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| series | Viruses |
| spelling | doaj.art-71b50dbe7d494af092ec0e1e3b64f3b32023-11-24T10:18:21ZengMDPI AGViruses1999-49152022-11-011411250910.3390/v14112509Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma MembranePrakash Raut0Bright Obeng1Hang Waters2Joshua Zimmerberg3Julie A. Gosse4Samuel T. Hess5Department of Physics and Astronomy, University of Maine, Orono, ME 04469-5709, USADepartment of Molecular and Biomedical Sciences, University of Maine, Orono, ME 04469-5735, USAEunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892-1855, USAEunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892-1855, USADepartment of Molecular and Biomedical Sciences, University of Maine, Orono, ME 04469-5735, USADepartment of Physics and Astronomy, University of Maine, Orono, ME 04469-5709, USAThe fully assembled influenza A virus (IAV) has on its surface the highest density of a single membrane protein found in nature—the glycoprotein hemagglutinin (HA) that mediates viral binding, entry, and assembly. HA clusters at the plasma membrane of infected cells, and the HA density (number of molecules per unit area) of these clusters correlates with the infectivity of the virus. Dense HA clusters are considered to mark the assembly site and ultimately lead to the budding of infectious IAV. The mechanism of spontaneous HA clustering, which occurs with or without other viral components, has not been elucidated. Using super-resolution fluorescence photoactivation localization microscopy (FPALM), we have previously shown that these HA clusters are interdependent on phosphatidylinositol 4,5-biphosphate (PIP2). Here, we show that the IAV matrix protein M1 co-clusters with PIP2, visualized using the pleckstrin homology domain. We find that cetylpyridinium chloride (CPC), which is a positively charged quaternary ammonium compound known for its antibacterial and antiviral properties at millimolar concentrations, disrupts M1 clustering and M1-PIP2 co-clustering at micromolar concentrations well below the critical micelle concentration (CMC). CPC also disrupts the co-clustering of M1 with HA at the plasma membrane, suggesting the role of host cell PIP2 clusters as scaffolds for gathering and concentrating M1 and HA to achieve their unusually high cluster densities in the IAV envelope.https://www.mdpi.com/1999-4915/14/11/2509influenza Asuper-resolution fluorescence microscopyhost–virus interactionsmatrix protein M1phosphoinositidesPIP2 |
| spellingShingle | Prakash Raut Bright Obeng Hang Waters Joshua Zimmerberg Julie A. Gosse Samuel T. Hess Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane Viruses influenza A super-resolution fluorescence microscopy host–virus interactions matrix protein M1 phosphoinositides PIP2 |
| title | Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane |
| title_full | Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane |
| title_fullStr | Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane |
| title_full_unstemmed | Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane |
| title_short | Phosphatidylinositol 4,5-Bisphosphate Mediates the Co-Distribution of Influenza A Hemagglutinin and Matrix Protein M1 at the Plasma Membrane |
| title_sort | phosphatidylinositol 4 5 bisphosphate mediates the co distribution of influenza a hemagglutinin and matrix protein m1 at the plasma membrane |
| topic | influenza A super-resolution fluorescence microscopy host–virus interactions matrix protein M1 phosphoinositides PIP2 |
| url | https://www.mdpi.com/1999-4915/14/11/2509 |
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