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

Gallo, G., Conceicao, C., Tsirigoti, C., Willett, B. , Graham, S. C. and Bailey, D. (2021) Application of error-prone PCR to functionally probe the morbillivirus Haemagglutinin protein. Journal of General Virology, 102(4), 001580. (doi: 10.1099/jgv.0.001580) (PMID:33739251) (PMCID:PMC8290269)

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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.

Item Type:Articles
Additional Information:This work was supported by the following grants to DB: a DHSC and UKRI BBSRC project grant (BB/R019843/1) and Institute Strategic Programme Grant (ISPG) to The Pirbright Institute (BBS/E/I/00007034, BBS/E/I/00007030 and BBS/E/I/00007039). SCG is a Sir Henry Dale Fellow, jointly funded by the Wellcome Trust and the Royal Society (098406/Z/12/B).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Willett, Professor Brian
Authors: Gallo, G., Conceicao, C., Tsirigoti, C., Willett, B., Graham, S. C., and Bailey, D.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Journal of General Virology
Publisher:Microbiology Society
ISSN:0022-1317
ISSN (Online):1465-2099
Published Online:19 March 2021
Copyright Holders:Copyright © 2021 The Authors
First Published:First published in Journal of General Virology 102(4): 001580
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
302172A One Health approach to pan-valent morbillivirus vaccinesBrian WillettBiotechnology and Biological Sciences Research Council (BBSRC)BB/R019843/1III - Centre for Virus Research