Norovirus evolution in immunodeficient mice reveals potentiated pathogenicity via a single nucleotide change in the viral capsid.

Forrest C Walker,Ebrahim Hassan,Timothy J Nice,Dylan Lawrence,Vincent R Graziano,Megan T Baldridge,Carla Blum-Johnston,Gowri Kalugotla,Jonathan J Miner,Alexa N Roth,Stephanie M Karst,Broc T Mccune,Stefan T Peterson,Larissa Lushniak,Rachel Rodgers,Yuhao Li,Lawrence A Schriefer,Craig B Wilen,Sang‐Hyun Lee ,Cassandra E Thompson

doi:10.1371/journal.ppat.1009402

Forrest C Walker, Ebrahim Hassan + Show 18 more

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https://doi.org/10.1371/journal.ppat.1009402

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Abstract

Interferons (IFNs) are key controllers of viral replication, with intact IFN responses suppressing virus growth and spread. Using the murine norovirus (MNoV) system, we show that IFNs exert selective pressure to limit the pathogenic evolutionary potential of this enteric virus. In animals lacking type I IFN signaling, the nonlethal MNoV strain CR6 rapidly acquired enhanced virulence via conversion of a single nucleotide. This nucleotide change resulted in amino acid substitution F514I in the viral capsid, which led to >10,000-fold higher replication in systemic organs including the brain. Pathogenicity was mediated by enhanced recruitment and infection of intestinal myeloid cells and increased extraintestinal dissemination of virus. Interestingly, the trade-off for this mutation was reduced fitness in an IFN-competent host, in which CR6 bearing F514I exhibited decreased intestinal replication and shedding. In an immunodeficient context, a spontaneous amino acid change can thus convert a relatively avirulent viral strain into a lethal pathogen.

Highlights

The investigation of viral evolution has long been vital to deciphering viral pathogenesis
To test whether CR6 accumulated viral mutation(s) that correlated with altered pathogenesis, we developed a method to efficiently deep sequence murine norovirus (MNoV) genomes (S2A Fig)
Using a reverse genetics system for MNoV [29], we introduced a single point mutation (T to A) at viral nucleotide 6595 of the CR6 infectious molecular clone, thereby generating the VP1-F514I mutation in the background of an otherwise unmodified CR6 genome, designated CR6F514I

Summary

Introduction

The investigation of viral evolution has long been vital to deciphering viral pathogenesis. This is true for RNA viruses, which evolve much more rapidly than DNA viruses [1]. In vivo viral evolution studies with influenza have provided key insights into the genetic basis of virulence and transmission [6,7]. Recent improvements in deep sequencing technology have made it increasingly accessible to probe inter- and intra-host evolution of viral genomes and have been applied extensively to influenza [8,9], HIV [10,11,12], and poliovirus [13,14]. Host interferons (IFNs) exert critical selective pressure to control viral infections, thereby regulating both viral evolution and transmission [15,16]

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