Rescue of Infectious Rotavirus Reassortants by a Reverse Genetics System Is Restricted by the Receptor-Binding Region of VP4.

Alexander Falkenhagen,Marno Huyzers,Alberdina A Van Dijk,Reimar Johne

doi:10.3390/v13030363

Alexander Falkenhagen, Marno Huyzers + Show 2 more

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https://doi.org/10.3390/v13030363

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Abstract

The rotavirus species A (RVA) capsid contains the spike protein VP4, which interacts with VP6 and VP7 and is involved in cellular receptor binding. The capsid encloses the genome consisting of eleven dsRNA segments. Reassortment events can result in novel strains with changed properties. Using a plasmid-based reverse genetics system based on simian RVA strain SA11, we previously showed that the rescue of viable reassortants containing a heterologous VP4-encoding genome segment was strain-dependent. In order to unravel the reasons for the reassortment restrictions, we designed here a series of plasmids encoding chimeric VP4s. Exchange of the VP4 domains interacting with VP6 and VP7 was not sufficient for rescue of viable viruses. In contrast, the exchange of fragments encoding the receptor-binding region of VP4 resulted in virus rescue. All parent strains and the rescued reassortants replicated efficiently in MA-104 cells used for virus propagation. In contrast, replication in BSR T7/5 cells used for plasmid transfection was only efficient for the SA11 strain, whereas the rescued reassortants replicated slowly, and the parent strains failing to produce reassortants did not replicate. While future research in this area is necessary, replication in BSR T7/5 cells may be one factor that affects the rescue of RVAs.

Highlights

IntroductionPublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations
We previously described the successful rescue of reassortant viruses containing VP4 from a diverse set of Rotavirus of species A (RVA) in the backbone of simian SA11 using a plasmid-based reverse genetics system [35,36]
Using a plasmid-based reverse genetics system, we have previously shown that VP4 from distantly related RVAs has a high reassortment potential with simian SA11

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Rotavirus of species A (RVA), a member of the family Reoviridae, is a non-enveloped virus with eleven dsRNA genome segments. Infections with RVA are the most common cause of severe gastroenteritis in young children worldwide and resulted in an estimated. 128,500 deaths among children younger than 5 years in 2016 [1]. RVA infections are widespread in wild animals and livestock, in which they can cause similar symptoms as in humans [2,3,4]. Interspecies transmission and reassortment events favor the emergence of novel RVA strains with new phenotypic properties, such as altered replication potential, immunogenic profile, or host range [5,6,7]

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