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Volume 105, Issue 8

The effect of pH on the structure of Bluetongue virus VP5 Open Access

Abstract

The unenveloped Bluetongue virus capsid comprises several structural layers, the inner two comprising a core, which assembles before addition of the outer proteins, VP2 and VP5. Two symmetric trimers of VP5 fit like pegs into two distinct pits on the core and undergo pH conformational changes in the context of the virus, associated with cell entry. Here we show that in isolation VP5 alone undergoes essentially the same changes with pH and confirm a helical transition, indicating that VP5 is a motor during cell entry. In the absence of VP5 the two pits on the core differ from each other, presumably due to the asymmetric underlying structure of VP3, the innermost capsid protein. On insertion of VP5 these pits become closely similar and remain similar at low pH whilst VP5 is present. This natural asymmetry presumably destabilises the attachment of VP5, facilitating ejection upon low pH, membrane penetration and cell entry.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Bluetongue virus , pH effect , virus capsid structure , virus entry , VP5 and X-ray crystallography
Funding
This study was supported by the:
  • Chinese Academy of Medical Sciences Initiative for Innovative Medicine (Award 2018-I2M-2-002)
    • Principle Award Recipient: DavidIan Stuart
  • Medical Research Council (Award MR/N00065X/1)
    • Principle Award Recipient: DavidIan Stuart
© 2024 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2024-08-20
2024-09-15
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References

  1. Hyatt AD, Eaton BT, Brookes SM. The release of bluetongue virus from infected cells and their superinfection by progeny virus. Virology 1989; 173:21–34 [View Article] [PubMed]
     [Google Scholar]
  2. Forzan M, Wirblich C, Roy P. A capsid protein of nonenveloped bluetongue virus exhibits membrane fusion activity. Proc Natl Acad Sci U S A 2004; 101:2100–2105 [View Article] [PubMed]
     [Google Scholar]
  3. Forzan M, Marsh M, Roy P. Bluetongue virus entry into cells. J Virol 2007; 81:4819–4827 [View Article] [PubMed]
     [Google Scholar]
  4. Hassan SH, Wirblich C, Forzan M, Roy P. Expression and functional characterization of bluetongue virus VP5 protein: role in cellular permeabilization. J Virol 2001; 75:8356–8367 [View Article] [PubMed]
     [Google Scholar]
  5. Zhang X, Patel A, Celma CC, Yu X, Roy P et al. Atomic model of a nonenveloped virus reveals pH sensors for a coordinated process of cell entry. Nat Struct Mol Biol 2016; 23:74–80 [View Article] [PubMed]
     [Google Scholar]
  6. Zhang X, Boyce M, Bhattacharya B, Zhang X, Schein S et al. Bluetongue virus coat protein VP2 contains sialic acid-binding domains, and VP5 resembles enveloped virus fusion proteins. Proc Natl Acad Sci USA 2010; 107:6292–6297 [View Article]
     [Google Scholar]
  7. Xia X, Wu W, Cui Y, Roy P, Zhou ZH. Bluetongue virus capsid protein VP5 perforates membranes at low endosomal pH during viral entry. Nat Microbiol 2021; 6:1424–1432 [View Article] [PubMed]
     [Google Scholar]
  8. Singh KP, Maan S, Samuel AR, Rao S, Meyer AJ et al. Phylogenetic analysis of bluetongue virus genome segment 6 (encoding VP5) from different serotypes. Vet Ital 2004; 40:479–483 [PubMed]
     [Google Scholar]
  9. Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R et al. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Res 2007; 35:e45 [View Article] [PubMed]
     [Google Scholar]
  10. Cronin CN, Lim KB, Rogers J. Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells. Protein Sci 2007; 16:2023–2029 [View Article] [PubMed]
     [Google Scholar]
  11. Aricescu AR, Assenberg R, Bill RM, Busso D, Chang VT et al. Eukaryotic expression: developments for structural proteomics. Acta Crystallogr D Biol Crystallogr 2006; 62:1114–1124 [View Article] [PubMed]
     [Google Scholar]
  12. Walter TS, Diprose JM, Mayo CJ, Siebold C, Pickford MG et al. A procedure for setting up high-throughput nanolitre crystallization experiments. Crystallization workflow for initial screening, automated storage, imaging and optimization. Acta Crystallogr D Biol Crystallogr 2005; 61:651–657 [View Article] [PubMed]
     [Google Scholar]
  13. Otwinowski Z, Minor W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol 1997; 276:307–326 [View Article] [PubMed]
     [Google Scholar]
  14. Sheldrick GM. A short history of SHELX. Acta Crystallogr A 2008; 64:112–122 [View Article] [PubMed]
     [Google Scholar]
  15. Adams PD, Afonine PV, Bunkóczi G, Chen VB, Davis IW et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr 2010; 66:213–221 [View Article] [PubMed]
     [Google Scholar]
  16. Emsley P, Cowtan K. Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol Crystallogr 2004; 60:2126–2132 [View Article] [PubMed]
     [Google Scholar]
  17. Stuart DI, Levine M, Muirhead H, Stammers DK. Crystal structure of cat muscle pyruvate kinase at A resolution of 2.6 A. J Mol Biol 1979; 134:109–142 [View Article] [PubMed]
     [Google Scholar]
  18. Wu W, Celma CC, Kerviel A, Roy P. Mapping the pH sensors critical for host cell entry by a complex nonenveloped virus. J Virol 2019; 93:e01897-18 [View Article] [PubMed]
     [Google Scholar]
  19. Braden B. The surveyor’s area formula. Coll Math J 1986; 17:326–337 [View Article]
     [Google Scholar]
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The effect of pH on the structure of Bluetongue virus VP5
J. Gen. Virol. 105, 002018 (2024); https://doi.org/10.1099/jgv.0.002018
/content/journal/jgv/10.1099/jgv.0.002018
/content/journal/jgv/10.1099/jgv.0.002018
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