Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions
The paramyxoviral phosphoprotein (P protein) is the non-catalytic subunit of the viral RNA polymerase, and coordinates many of the molecular interactions required for RNA synthesis. All paramyxoviral P proteins oligomerize via a centrally located coiled-coil that is connected to a downstream binding...
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MDPI AG
2021-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/9/1737 |
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| author | Melissa N. Webby Nicole Herr Esther M. M. Bulloch Michael Schmitz Jeremy R. Keown David C. Goldstone Richard L. Kingston |
| author_facet | Melissa N. Webby Nicole Herr Esther M. M. Bulloch Michael Schmitz Jeremy R. Keown David C. Goldstone Richard L. Kingston |
| author_sort | Melissa N. Webby |
| collection | DOAJ |
| description | The paramyxoviral phosphoprotein (P protein) is the non-catalytic subunit of the viral RNA polymerase, and coordinates many of the molecular interactions required for RNA synthesis. All paramyxoviral P proteins oligomerize via a centrally located coiled-coil that is connected to a downstream binding domain by a dynamic linker. The C-terminal region of the P protein coordinates interactions between the catalytic subunit of the polymerase, and the viral nucleocapsid housing the genomic RNA. The inherent flexibility of the linker is believed to facilitate polymerase translocation. Here we report biophysical and structural characterization of the C-terminal region of the P protein from Menangle virus (MenV), a bat-borne paramyxovirus with zoonotic potential. The MenV P protein is tetrameric but can dissociate into dimers at sub-micromolar protein concentrations. The linker is globally disordered and can be modeled effectively as a worm-like chain. However, NMR analysis suggests very weak local preferences for alpha-helical and extended beta conformation exist within the linker. At the interface between the disordered linker and the structured C-terminal binding domain, a gradual disorder-to-order transition occurs, with X-ray crystallographic analysis revealing a dynamic interfacial structure that wraps the surface of the binding domain. |
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| id | doaj.art-fb1fa7aa7a7d4673b947e3430ec172ed |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T07:09:11Z |
| publishDate | 2021-08-01 |
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| spelling | doaj.art-fb1fa7aa7a7d4673b947e3430ec172ed2023-11-22T15:37:13ZengMDPI AGViruses1999-49152021-08-01139173710.3390/v13091737Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered RegionsMelissa N. Webby0Nicole Herr1Esther M. M. Bulloch2Michael Schmitz3Jeremy R. Keown4David C. Goldstone5Richard L. Kingston6School of Biological Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Biological Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Biological Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Chemical Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Biological Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Biological Sciences, University of Auckland, Auckland 1010, New ZealandSchool of Biological Sciences, University of Auckland, Auckland 1010, New ZealandThe paramyxoviral phosphoprotein (P protein) is the non-catalytic subunit of the viral RNA polymerase, and coordinates many of the molecular interactions required for RNA synthesis. All paramyxoviral P proteins oligomerize via a centrally located coiled-coil that is connected to a downstream binding domain by a dynamic linker. The C-terminal region of the P protein coordinates interactions between the catalytic subunit of the polymerase, and the viral nucleocapsid housing the genomic RNA. The inherent flexibility of the linker is believed to facilitate polymerase translocation. Here we report biophysical and structural characterization of the C-terminal region of the P protein from Menangle virus (MenV), a bat-borne paramyxovirus with zoonotic potential. The MenV P protein is tetrameric but can dissociate into dimers at sub-micromolar protein concentrations. The linker is globally disordered and can be modeled effectively as a worm-like chain. However, NMR analysis suggests very weak local preferences for alpha-helical and extended beta conformation exist within the linker. At the interface between the disordered linker and the structured C-terminal binding domain, a gradual disorder-to-order transition occurs, with X-ray crystallographic analysis revealing a dynamic interfacial structure that wraps the surface of the binding domain.https://www.mdpi.com/1999-4915/13/9/1737ParamyxovirusesRNA-dependent RNA polymerasepararubulavirusesintrinsically disordered proteinsprotein self-associationconformational exchange |
| spellingShingle | Melissa N. Webby Nicole Herr Esther M. M. Bulloch Michael Schmitz Jeremy R. Keown David C. Goldstone Richard L. Kingston Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions Viruses Paramyxoviruses RNA-dependent RNA polymerase pararubulaviruses intrinsically disordered proteins protein self-association conformational exchange |
| title | Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions |
| title_full | Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions |
| title_fullStr | Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions |
| title_full_unstemmed | Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions |
| title_short | Structural Analysis of the Menangle Virus P Protein Reveals a Soft Boundary between Ordered and Disordered Regions |
| title_sort | structural analysis of the menangle virus p protein reveals a soft boundary between ordered and disordered regions |
| topic | Paramyxoviruses RNA-dependent RNA polymerase pararubulaviruses intrinsically disordered proteins protein self-association conformational exchange |
| url | https://www.mdpi.com/1999-4915/13/9/1737 |
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