A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles
ABSTRACT Respiratory syncytial virus (RSV) is a single-stranded RNA virus in the Paramyxoviridae family that assembles into filamentous structures at the apical surface of polarized epithelial cells. These filaments contain viral genomic RNA and structural proteins, including the fusion (F) protein,...
Guardado en:
| Autores principales: | , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
American Society for Microbiology
2012
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/415b299e036d4dc4b9541181cd84fc96 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:415b299e036d4dc4b9541181cd84fc96 |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:415b299e036d4dc4b9541181cd84fc962021-11-15T15:39:02ZA Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles10.1128/mBio.00270-112150-7511https://doaj.org/article/415b299e036d4dc4b9541181cd84fc962012-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00270-11https://doaj.org/toc/2150-7511ABSTRACT Respiratory syncytial virus (RSV) is a single-stranded RNA virus in the Paramyxoviridae family that assembles into filamentous structures at the apical surface of polarized epithelial cells. These filaments contain viral genomic RNA and structural proteins, including the fusion (F) protein, matrix (M) protein, nucleoprotein (N), and phosphoprotein (P), while excluding F-actin. It is known that the F protein cytoplasmic tail (FCT) is necessary for filament formation, but the mechanism by which the FCT mediates assembly into filaments is not clear. We hypothesized that the FCT is necessary for interactions with other viral proteins in order to form filaments. In order to test this idea, we expressed the F protein with cytoplasmic tail (CT) truncations or specific point mutations and determined the abilities of these variant F proteins to form filaments independent of viral infection when coexpressed with M, N, and P. Deletion of the terminal three FCT residues (amino acids Phe-Ser-Asn) or mutation of the Phe residue resulted in a loss of filament formation but did not affect F-protein expression or trafficking to the cell surface. Filament formation could be restored by addition of residues Phe-Ser-Asn to an FCT deletion mutant and was unaffected by mutations to Ser or Asn residues. Second, deletion of residues Phe-Ser-Asn or mutation of the Phe residue resulted in a loss of M, N, and P incorporation into virus-like particles. These data suggest that a C-terminal Phe residue in the FCT mediates assembly through incorporation of internal virion proteins into virus filaments at the cell surface. IMPORTANCE Respiratory syncytial virus (RSV) is a leading cause of bronchiolitis and pneumonia in infants and the elderly worldwide. There is no licensed RSV vaccine and only limited therapeutics for use in infected patients. Many aspects of the RSV life cycle have been studied, but the mechanisms that drive RSV assembly at the cell surface are not well understood. This study provides evidence that a specific residue in the RSV fusion protein cytoplasmic tail coordinates assembly into viral filaments by mediating the incorporation of internal virion proteins. Understanding the mechanisms that drive RSV assembly could lead to targeted development of novel antiviral drugs. Moreover, since RSV exits infected cells in an ESCRT (endosomal sorting complexes required for transport)-independent manner, these studies may contribute new knowledge about a general strategy by which ESCRT-independent viruses mediate outward bud formation using viral protein-mediated mechanisms during assembly and budding.Fyza Y. ShaikhReagan G. CoxAaron W. LiflandAnne L. HotardJohn V. WilliamsMartin L. MoorePhilip J. SantangeloJames E. CroweAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 1 (2012) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Microbiology QR1-502 |
| spellingShingle |
Microbiology QR1-502 Fyza Y. Shaikh Reagan G. Cox Aaron W. Lifland Anne L. Hotard John V. Williams Martin L. Moore Philip J. Santangelo James E. Crowe A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| description |
ABSTRACT Respiratory syncytial virus (RSV) is a single-stranded RNA virus in the Paramyxoviridae family that assembles into filamentous structures at the apical surface of polarized epithelial cells. These filaments contain viral genomic RNA and structural proteins, including the fusion (F) protein, matrix (M) protein, nucleoprotein (N), and phosphoprotein (P), while excluding F-actin. It is known that the F protein cytoplasmic tail (FCT) is necessary for filament formation, but the mechanism by which the FCT mediates assembly into filaments is not clear. We hypothesized that the FCT is necessary for interactions with other viral proteins in order to form filaments. In order to test this idea, we expressed the F protein with cytoplasmic tail (CT) truncations or specific point mutations and determined the abilities of these variant F proteins to form filaments independent of viral infection when coexpressed with M, N, and P. Deletion of the terminal three FCT residues (amino acids Phe-Ser-Asn) or mutation of the Phe residue resulted in a loss of filament formation but did not affect F-protein expression or trafficking to the cell surface. Filament formation could be restored by addition of residues Phe-Ser-Asn to an FCT deletion mutant and was unaffected by mutations to Ser or Asn residues. Second, deletion of residues Phe-Ser-Asn or mutation of the Phe residue resulted in a loss of M, N, and P incorporation into virus-like particles. These data suggest that a C-terminal Phe residue in the FCT mediates assembly through incorporation of internal virion proteins into virus filaments at the cell surface. IMPORTANCE Respiratory syncytial virus (RSV) is a leading cause of bronchiolitis and pneumonia in infants and the elderly worldwide. There is no licensed RSV vaccine and only limited therapeutics for use in infected patients. Many aspects of the RSV life cycle have been studied, but the mechanisms that drive RSV assembly at the cell surface are not well understood. This study provides evidence that a specific residue in the RSV fusion protein cytoplasmic tail coordinates assembly into viral filaments by mediating the incorporation of internal virion proteins. Understanding the mechanisms that drive RSV assembly could lead to targeted development of novel antiviral drugs. Moreover, since RSV exits infected cells in an ESCRT (endosomal sorting complexes required for transport)-independent manner, these studies may contribute new knowledge about a general strategy by which ESCRT-independent viruses mediate outward bud formation using viral protein-mediated mechanisms during assembly and budding. |
| format |
article |
| author |
Fyza Y. Shaikh Reagan G. Cox Aaron W. Lifland Anne L. Hotard John V. Williams Martin L. Moore Philip J. Santangelo James E. Crowe |
| author_facet |
Fyza Y. Shaikh Reagan G. Cox Aaron W. Lifland Anne L. Hotard John V. Williams Martin L. Moore Philip J. Santangelo James E. Crowe |
| author_sort |
Fyza Y. Shaikh |
| title |
A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| title_short |
A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| title_full |
A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| title_fullStr |
A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| title_full_unstemmed |
A Critical Phenylalanine Residue in the Respiratory Syncytial Virus Fusion Protein Cytoplasmic Tail Mediates Assembly of Internal Viral Proteins into Viral Filaments and Particles |
| title_sort |
critical phenylalanine residue in the respiratory syncytial virus fusion protein cytoplasmic tail mediates assembly of internal viral proteins into viral filaments and particles |
| publisher |
American Society for Microbiology |
| publishDate |
2012 |
| url |
https://doaj.org/article/415b299e036d4dc4b9541181cd84fc96 |
| work_keys_str_mv |
AT fyzayshaikh acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT reagangcox acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT aaronwlifland acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT annelhotard acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT johnvwilliams acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT martinlmoore acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT philipjsantangelo acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT jamesecrowe acriticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT fyzayshaikh criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT reagangcox criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT aaronwlifland criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT annelhotard criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT johnvwilliams criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT martinlmoore criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT philipjsantangelo criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles AT jamesecrowe criticalphenylalanineresidueintherespiratorysyncytialvirusfusionproteincytoplasmictailmediatesassemblyofinternalviralproteinsintoviralfilamentsandparticles |
| _version_ |
1718427760034054144 |