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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
2024-04-27
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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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