Related Experiment Video
Updated: Nov 12, 2025
Engineering Antiviral Agents via Surface Plasmon Resonance
Published on: June 14, 2022
Application of error-prone PCR to functionally probe the morbillivirus Haemagglutinin protein
Giulia Gallo1, Carina Conceicao1, Christina Tsirigoti1
1The Pirbright Institute, Guildford, Surrey, GU24 0NF, UK.
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.
Insights
Peste des petits ruminants virus (PPRV) Haemagglutinin (H) protein interactions with SLAMF1 are functionally conserved across morbilliviruses. This finding, identified through random mutagenesis and structural modeling, confirms a shared viral entry mechanism.
Area of Science:
- Virology
- Structural Biology
- Molecular Biology
Background:
- Morbilliviruses, a genus of enveloped viruses, utilize specific protein receptors like SLAMF1 or Nectin-4 for host cell entry.
- The viral Haemagglutinin (H) protein initiates receptor binding, while the Fusion (F) protein mediates membrane fusion.
- Existing crystal structures of measles virus H bound to SLAMF1 or Nectin-4 provide insights, but conservation across the genus needs investigation.
Purpose of the Study:
- To investigate the functional conservation of the SLAMF1 receptor binding site on the Haemagglutinin (H) protein of peste des petits ruminants virus (PPRV).
- To determine if the interaction mechanism between PPRV H and SLAMF1 is conserved across the morbillivirus genus.
Main Methods:
- Random mutagenesis of the PPRV H protein targeting the putative SLAMF1 receptor binding site using error-prone PCR.
- Assessing the impact of mutations on virus-induced cell-cell fusion.
- Structural modeling of the interaction between PPRV H and ovine SLAMF1.
Main Results:
- Identified specific mutations in PPRV H that significantly inhibited virus-induced cell-cell fusion, indicating disruption of SLAMF1 binding.
- Structural modeling supported the functional data, highlighting a conserved interaction region between PPRV H and SLAMF1.
- The findings suggest that the SLAMF1 binding site on the H protein is functionally conserved throughout the morbillivirus genus.
Conclusions:
- The receptor binding site for SLAMF1 on the morbillivirus Haemagglutinin protein is functionally conserved across the genus.
- Error-prone PCR is a valuable method for functionally characterizing viral protein domains involved in host cell entry.
- This study enhances understanding of morbillivirus-host interactions and provides a basis for antiviral strategies.
