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 |
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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 > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research |
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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