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Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion
There has been resurgence in determining the role of host metabolism in viral infection yet deciphering how the metabolic state of single cells affects viral entry and fusion remains unknown. Here, we have developed a novel assay multiplexing genetically-encoded biosensors with single virus tracking...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7055913/ https://www.ncbi.nlm.nih.gov/pubmed/32084246 http://dx.doi.org/10.1371/journal.ppat.1008359 |
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author | Coomer, Charles A. Carlon-Andres, Irene Iliopoulou, Maro Dustin, Michael L. Compeer, Ewoud B. Compton, Alex A. Padilla-Parra, Sergi |
author_facet | Coomer, Charles A. Carlon-Andres, Irene Iliopoulou, Maro Dustin, Michael L. Compeer, Ewoud B. Compton, Alex A. Padilla-Parra, Sergi |
author_sort | Coomer, Charles A. |
collection | PubMed |
description | There has been resurgence in determining the role of host metabolism in viral infection yet deciphering how the metabolic state of single cells affects viral entry and fusion remains unknown. Here, we have developed a novel assay multiplexing genetically-encoded biosensors with single virus tracking (SVT) to evaluate the influence of global metabolic processes on the success rate of virus entry in single cells. We found that cells with a lower ATP:ADP ratio prior to virus addition were less permissive to virus fusion and infection. These results indicated a relationship between host metabolic state and the likelihood for virus-cell fusion to occur. SVT revealed that HIV-1 virions were arrested at hemifusion in glycolytically-inactive cells. Interestingly, cells acutely treated with glycolysis inhibitor 2-deoxyglucose (2-DG) become resistant to virus infection and also display less surface membrane cholesterol. Addition of cholesterol in these in glycolytically-inactive cells rescued the virus entry block at hemifusion and enabled completion of HIV-1 fusion. Further investigation with FRET-based membrane tension and membrane order reporters revealed a link between host cell glycolytic activity and host membrane order and tension. Indeed, cells treated with 2-DG possessed lower plasma membrane lipid order and higher tension values, respectively. Our novel imaging approach that combines lifetime imaging (FLIM) and SVT revealed not only changes in plasma membrane tension at the point of viral fusion, but also that HIV is less likely to enter cells at areas of higher membrane tension. We therefore have identified a connection between host cell glycolytic activity and membrane tension that influences HIV-1 fusion in real-time at the single-virus fusion level in live cells. |
format | Online Article Text |
id | pubmed-7055913 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-70559132020-03-13 Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion Coomer, Charles A. Carlon-Andres, Irene Iliopoulou, Maro Dustin, Michael L. Compeer, Ewoud B. Compton, Alex A. Padilla-Parra, Sergi PLoS Pathog Research Article There has been resurgence in determining the role of host metabolism in viral infection yet deciphering how the metabolic state of single cells affects viral entry and fusion remains unknown. Here, we have developed a novel assay multiplexing genetically-encoded biosensors with single virus tracking (SVT) to evaluate the influence of global metabolic processes on the success rate of virus entry in single cells. We found that cells with a lower ATP:ADP ratio prior to virus addition were less permissive to virus fusion and infection. These results indicated a relationship between host metabolic state and the likelihood for virus-cell fusion to occur. SVT revealed that HIV-1 virions were arrested at hemifusion in glycolytically-inactive cells. Interestingly, cells acutely treated with glycolysis inhibitor 2-deoxyglucose (2-DG) become resistant to virus infection and also display less surface membrane cholesterol. Addition of cholesterol in these in glycolytically-inactive cells rescued the virus entry block at hemifusion and enabled completion of HIV-1 fusion. Further investigation with FRET-based membrane tension and membrane order reporters revealed a link between host cell glycolytic activity and host membrane order and tension. Indeed, cells treated with 2-DG possessed lower plasma membrane lipid order and higher tension values, respectively. Our novel imaging approach that combines lifetime imaging (FLIM) and SVT revealed not only changes in plasma membrane tension at the point of viral fusion, but also that HIV is less likely to enter cells at areas of higher membrane tension. We therefore have identified a connection between host cell glycolytic activity and membrane tension that influences HIV-1 fusion in real-time at the single-virus fusion level in live cells. Public Library of Science 2020-02-21 /pmc/articles/PMC7055913/ /pubmed/32084246 http://dx.doi.org/10.1371/journal.ppat.1008359 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 (https://creativecommons.org/publicdomain/zero/1.0/) public domain dedication. |
spellingShingle | Research Article Coomer, Charles A. Carlon-Andres, Irene Iliopoulou, Maro Dustin, Michael L. Compeer, Ewoud B. Compton, Alex A. Padilla-Parra, Sergi Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title | Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title_full | Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title_fullStr | Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title_full_unstemmed | Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title_short | Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion |
title_sort | single-cell glycolytic activity regulates membrane tension and hiv-1 fusion |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7055913/ https://www.ncbi.nlm.nih.gov/pubmed/32084246 http://dx.doi.org/10.1371/journal.ppat.1008359 |
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