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. Associa...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-09-01
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| Series: | Viruses |
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| Online Access: | http://www.mdpi.com/1999-4915/10/10/535 |
| _version_ | 1818317475929915392 |
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| author | Shadi Shahriari Ke-jun Wei Reena Ghildyal |
| author_facet | Shadi Shahriari Ke-jun Wei Reena Ghildyal |
| author_sort | Shadi Shahriari |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-13T09:37:55Z |
| format | Article |
| id | doaj.art-e9be95bc43c54345b082cba45608bbc8 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-13T09:37:55Z |
| publishDate | 2018-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e9be95bc43c54345b082cba45608bbc82022-12-21T23:52:18ZengMDPI AGViruses1999-49152018-09-01101053510.3390/v10100535v10100535Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell CultureShadi Shahriari0Ke-jun Wei1Reena Ghildyal2Centre for Research in Therapeutic Solutions, Faculty of Science and Technology, University of Canberra, Canberra ACT 2617, AustraliaCentre for Research in Therapeutic Solutions, Faculty of Science and Technology, University of Canberra, Canberra ACT 2617, AustraliaCentre for Research in Therapeutic Solutions, Faculty of Science and Technology, University of Canberra, Canberra ACT 2617, AustraliaThe 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.http://www.mdpi.com/1999-4915/10/10/535actin cytoskeletonvirus transportrespiratory syncytial virusmatrix protein |
| spellingShingle | Shadi Shahriari Ke-jun Wei Reena Ghildyal Respiratory Syncytial Virus Matrix (M) Protein Interacts with Actin In Vitro and in Cell Culture Viruses actin cytoskeleton virus transport respiratory syncytial virus matrix protein |
| 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 | actin cytoskeleton virus transport respiratory syncytial virus matrix protein |
| url | http://www.mdpi.com/1999-4915/10/10/535 |
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