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Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture
The virus–host protein interactions that underlie respiratory syncytial virus (RSV) assembly are still not completely defined, despite almost 60 years of research. RSV buds from the apical surface of infected cells, once virion components have been transported to the budding sites. Association of RS...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213044/ https://www.ncbi.nlm.nih.gov/pubmed/30274351 http://dx.doi.org/10.3390/v10100535 |
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author | Shahriari, Shadi Wei, Ke-jun Ghildyal, Reena |
author_facet | Shahriari, Shadi Wei, Ke-jun Ghildyal, Reena |
author_sort | Shahriari, Shadi |
collection | PubMed |
description | The virus–host protein interactions that underlie respiratory syncytial virus (RSV) assembly are still not completely defined, despite almost 60 years of research. RSV buds from the apical surface of infected cells, once virion components have been transported to the budding sites. Association of RSV matrix (M) protein with the actin cytoskeleton may play a role in facilitating this transport. We have investigated the interaction of M with actin in vitro and cell culture. Purified wildtype RSV M protein was found to bind directly to polymerized actin in vitro. Vero cells were transfected to express full-length M (1–256) as a green fluorescent protein-(GFP) tagged protein, followed by treatment with the microfilament destabilizer, cytochalasin D. Destabilization of the microfilament network resulted in mislocalization of full-length M, from mostly cytoplasmic to diffused across both cytoplasm and nucleus, suggesting that M interacts with microfilaments in this system. Importantly, treatment of RSV-infected cells with cytochalasin D results in lower infectious virus titers, as well as mislocalization of M to the nucleus. Finally, using deletion mutants of M in a transfected cell system, we show that both the N- and C-terminus of the protein are required for the interaction. Together, our data suggest a possible role for M–actin interaction in transporting virion components in the infected cell. |
format | Online Article Text |
id | pubmed-6213044 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-62130442018-11-09 Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture Shahriari, Shadi Wei, Ke-jun Ghildyal, Reena Viruses Article The virus–host protein interactions that underlie respiratory syncytial virus (RSV) assembly are still not completely defined, despite almost 60 years of research. RSV buds from the apical surface of infected cells, once virion components have been transported to the budding sites. Association of RSV matrix (M) protein with the actin cytoskeleton may play a role in facilitating this transport. We have investigated the interaction of M with actin in vitro and cell culture. Purified wildtype RSV M protein was found to bind directly to polymerized actin in vitro. Vero cells were transfected to express full-length M (1–256) as a green fluorescent protein-(GFP) tagged protein, followed by treatment with the microfilament destabilizer, cytochalasin D. Destabilization of the microfilament network resulted in mislocalization of full-length M, from mostly cytoplasmic to diffused across both cytoplasm and nucleus, suggesting that M interacts with microfilaments in this system. Importantly, treatment of RSV-infected cells with cytochalasin D results in lower infectious virus titers, as well as mislocalization of M to the nucleus. Finally, using deletion mutants of M in a transfected cell system, we show that both the N- and C-terminus of the protein are required for the interaction. Together, our data suggest a possible role for M–actin interaction in transporting virion components in the infected cell. MDPI 2018-09-30 /pmc/articles/PMC6213044/ /pubmed/30274351 http://dx.doi.org/10.3390/v10100535 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Shahriari, Shadi Wei, Ke-jun Ghildyal, Reena Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title | Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title_full | Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title_fullStr | Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title_full_unstemmed | Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title_short | Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture |
title_sort | respiratory syncytial virus matrix (m) protein interacts with actin in vitro and in cell culture |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213044/ https://www.ncbi.nlm.nih.gov/pubmed/30274351 http://dx.doi.org/10.3390/v10100535 |
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