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Volume 102, Issue 4

Application of error-prone PCR to functionally probe the morbillivirus Haemagglutinin protein Open Access

Abstract

The enveloped morbilliviruses utilise conserved proteinaceous receptors to enter host cells: SLAMF1 or Nectin-4. Receptor binding is initiated by the viral attachment protein Haemagglutinin (H), with the viral Fusion protein (F) driving membrane fusion. Crystal structures of the prototypic morbillivirus measles virus H with either SLAMF1 or Nectin-4 are available and have served as the basis for improved understanding of this interaction. However, whether these interactions remain conserved throughout the morbillivirus genus requires further characterisation. Using a random mutagenesis approach, based on error-prone PCR, we targeted the putative receptor binding site for SLAMF1 interaction on peste des petits ruminants virus (PPRV) H, identifying mutations that inhibited virus-induced cell-cell fusion. These data, combined with structural modelling of the PPRV H and ovine SLAMF1 interaction, indicate this region is functionally conserved across all morbilliviruses. Error-prone PCR provides a powerful tool for functionally characterising functional domains within viral proteins.

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  • Accepted:
  • Published Online:
Keyword(s): directed evolution , epPCR , morbillivirus , peste des petits ruminants virus , viral entry and viral evolution
Funding
This study was supported by the:
  • Wellcome Trust (Award 098406/Z/12/B)
    • Principle Award Recipient: StephenGraham
  • Biotechnology and Biological Sciences Research Council (Award BBS/E/I/00007039)
    • Principle Award Recipient: DalanBailey
  • Biotechnology and Biological Sciences Research Council (Award BBS/E/I/00007030)
    • Principle Award Recipient: StephenGraham
  • Biotechnology and Biological Sciences Research Council (Award BBS/E/I/00007034)
    • Principle Award Recipient: DalanBailey
  • Biotechnology and Biological Sciences Research Council (Award BB/R019843/1)
    • Principle Award Recipient: DalanBailey
© 2021 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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2021-04-08
2024-12-06
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References

  1. Tatsuo H, Ono N, Tanaka K, Yanagi Y. Slam (CDw150) is a cellular receptor for measles virus. Nature 2000; 406:893–897 [View Article][PubMed]
     [Google Scholar]
  2. Mühlebach MD, Mateo M, Sinn PL, Prüfer S, Uhlig KM et al. Adherens junction protein Nectin-4 is the epithelial receptor for measles virus. Nature 2011; 480:530–533 [View Article][PubMed]
     [Google Scholar]
  3. Noyce RS, Bondre DG, Ha MN, Lin L-T, Sisson G et al. Tumor cell marker PVRL4 (nectin 4) is an epithelial cell receptor for measles virus. PLoS Pathog 2011; 7:e1002240 [View Article][PubMed]
     [Google Scholar]
  4. Birch J, Juleff N, Heaton MP, Kalbfleisch T, Kijas J et al. Characterization of ovine Nectin-4, a novel peste des petits ruminants virus receptor. J Virol 2013; 87:4756–4761 [View Article][PubMed]
     [Google Scholar]
  5. Fukuhara H, Mwaba MH, Maenaka K. Structural characteristics of measles virus entry. Curr Opin Virol 2020; 41:52–58 [View Article][PubMed]
     [Google Scholar]
  6. Navaratnarajah CK, Generous AR, Yousaf I, Cattaneo R. Receptor-Mediated cell entry of paramyxoviruses: mechanisms, and consequences for tropism and pathogenesis. J Biol Chem 2020; 295:2771–2786 [View Article][PubMed]
     [Google Scholar]
  7. Bieringer M, Han JW, Kendl S, Khosravi M, Plattet P et al. Experimental adaptation of wild-type canine distemper virus (CDV) to the human entry receptor CD150. PLoS One 2013; 8:e57488 [View Article][PubMed]
     [Google Scholar]
  8. Abdullah N, Kelly JT, Graham SC, Birch J, Gonçalves-Carneiro D et al. Structure-Guided identification of a nonhuman morbillivirus with zoonotic potential. J Virol 2018; 92: 01 12 2018 [View Article][PubMed]
     [Google Scholar]
  9. Hashiguchi T, Ose T, Kubota M, Maita N, Kamishikiryo J et al. Structure of the measles virus hemagglutinin bound to its cellular receptor SLAM. Nat Struct Mol Biol 2011; 18:135–141 [View Article][PubMed]
     [Google Scholar]
  10. Fukuhara H, Ito Y, Sako M, Kajikawa M, Yoshida K et al. Specificity of morbillivirus hemagglutinins to recognize SLAM of different species. Viruses 2019; 11:761 19 08 2019 [View Article][PubMed]
     [Google Scholar]
  11. Navaratnarajah CK, Rosemarie Q, Cattaneo R. A structurally unresolved head segment of defined length favors proper measles virus hemagglutinin tetramerization and efficient membrane fusion triggering. J Virol 2016; 90:68–75 [View Article][PubMed]
     [Google Scholar]
  12. Zhang X, Lu G, Qi J, Li Y, He Y et al. Structure of measles virus hemagglutinin bound to its epithelial receptor Nectin-4. Nat Struct Mol Biol 2013; 20:67–72 [View Article][PubMed]
     [Google Scholar]
  13. McCullum EO, Williams BAR, Zhang J, Chaput JC. Random mutagenesis by error-prone PCR. Methods Mol Biol 2010; 634:103–109 [View Article][PubMed]
     [Google Scholar]
  14. Bailey D, Banyard A, Dash P, Ozkul A, Barrett T. Full genome sequence of peste des petits ruminants virus, a member of the morbillivirus genus. Virus Res 2005; 110:119–124 [View Article][PubMed]
     [Google Scholar]
  15. Arias A, Thorne L, Goodfellow I. Favipiravir elicits antiviral mutagenesis during virus replication in vivo . elife 2014; 3:e03679 [View Article][PubMed]
     [Google Scholar]
  16. Ishikawa H, Meng F, Kondo N, Iwamoto A, Matsuda Z. Generation of a dual-functional split-reporter protein for monitoring membrane fusion using self-associating split GFP. Protein Eng Des Sel 2012; 25:813–820 [View Article][PubMed]
     [Google Scholar]
  17. Wang Z, Moult J, SNPs MJ. Snps, protein structure, and disease. Hum Mutat 2001; 17:263–270 [View Article][PubMed]
     [Google Scholar]
  18. Navaratnarajah CK, Kumar S, Generous A, Apte-Sengupta S, Mateo M et al. The measles virus hemagglutinin stalk: structures and functions of the central fusion activation and membrane-proximal segments. J Virol 2014; 88:6158–6167 [View Article][PubMed]
     [Google Scholar]
  19. Starr TN, Greaney AJ, Hilton SK, Ellis D, Crawford KHD et al. Deep mutational scanning of SARS-CoV-2 receptor binding domain reveals constraints on folding and ACE2 binding. Cell 2020; 182:e12201295–1310 [View Article][PubMed]
     [Google Scholar]
  20. Koerber JT, Maheshri N, Kaspar BK, Schaffer DV. Construction of diverse adeno-associated viral libraries for directed evolution of enhanced gene delivery vehicles. Nat Protoc 2006; 1:701–706 [View Article][PubMed]
     [Google Scholar]
  21. Tee KL, Wong TS. Polishing the craft of genetic diversity creation in directed evolution. Biotechnol Adv 2013; 31:1707–1721 [View Article][PubMed]
     [Google Scholar]
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Application of error-prone PCR to functionally probe the morbillivirus Haemagglutinin protein
J. Gen. Virol. 102, 001580 (2021); https://doi.org/10.1099/jgv.0.001580
/content/journal/jgv/10.1099/jgv.0.001580
/content/journal/jgv/10.1099/jgv.0.001580
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