Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production
ABSTRACT Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how non...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020-10-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.01869-20 |
| _version_ | 1818394795101388800 |
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| author | Ricardo S. Cardoso Lucas Alves Tavares Bruna Lais S. Jesus Miria F. Criado Andreia Nogueira de Carvalho Juliano de Paula Souza Sukhmani Bedi Marcos Michel de Souza Maria Lucia Silva Guilherme Pauperio Lanfredi Brenda Cristina Vitti Orlando Bonito Scudero Vitor Marcel Faça Akira Ono Armando M. Ventura Luis Lamberti P. daSilva Eurico Arruda |
| author_facet | Ricardo S. Cardoso Lucas Alves Tavares Bruna Lais S. Jesus Miria F. Criado Andreia Nogueira de Carvalho Juliano de Paula Souza Sukhmani Bedi Marcos Michel de Souza Maria Lucia Silva Guilherme Pauperio Lanfredi Brenda Cristina Vitti Orlando Bonito Scudero Vitor Marcel Faça Akira Ono Armando M. Ventura Luis Lamberti P. daSilva Eurico Arruda |
| author_sort | Ricardo S. Cardoso |
| collection | DOAJ |
| description | ABSTRACT Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites. IMPORTANCE The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process. |
| first_indexed | 2024-12-14T06:06:53Z |
| format | Article |
| id | doaj.art-677e053382274242a051b92585a6cb7e |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-14T06:06:53Z |
| publishDate | 2020-10-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-677e053382274242a051b92585a6cb7e2022-12-21T23:14:15ZengAmerican Society for MicrobiologymBio2150-75112020-10-0111510.1128/mBio.01869-20Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral ProductionRicardo S. Cardoso0Lucas Alves Tavares1Bruna Lais S. Jesus2Miria F. Criado3Andreia Nogueira de Carvalho4Juliano de Paula Souza5Sukhmani Bedi6Marcos Michel de Souza7Maria Lucia Silva8Guilherme Pauperio Lanfredi9Brenda Cristina Vitti10Orlando Bonito Scudero11Vitor Marcel Faça12Akira Ono13Armando M. Ventura14Luis Lamberti P. daSilva15Eurico Arruda16Department of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Microbiology and Immunology, University Michigan Medical School, Ann Arbor, Michigan, USADepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Biochemistry, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartment of Biochemistry, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Microbiology and Immunology, University Michigan Medical School, Ann Arbor, Michigan, USADepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilDepartment of Cell and Molecular Biology, University of São Paulo School of Medicine, Ribeirao Preto, BrazilABSTRACT Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites. IMPORTANCE The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process.https://journals.asm.org/doi/10.1128/mBio.01869-20human respiratory syncytial virusparamyxovirusprotein traffickingvirus-host interactions |
| spellingShingle | Ricardo S. Cardoso Lucas Alves Tavares Bruna Lais S. Jesus Miria F. Criado Andreia Nogueira de Carvalho Juliano de Paula Souza Sukhmani Bedi Marcos Michel de Souza Maria Lucia Silva Guilherme Pauperio Lanfredi Brenda Cristina Vitti Orlando Bonito Scudero Vitor Marcel Faça Akira Ono Armando M. Ventura Luis Lamberti P. daSilva Eurico Arruda Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production mBio human respiratory syncytial virus paramyxovirus protein trafficking virus-host interactions |
| title | Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production |
| title_full | Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production |
| title_fullStr | Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production |
| title_full_unstemmed | Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production |
| title_short | Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production |
| title_sort | host retromer protein sorting nexin 2 interacts with human respiratory syncytial virus structural proteins and is required for efficient viral production |
| topic | human respiratory syncytial virus paramyxovirus protein trafficking virus-host interactions |
| url | https://journals.asm.org/doi/10.1128/mBio.01869-20 |
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