The Amino Acid Substitution Q65H in the 2C Protein of Swine Vesicular Disease Virus Confers Resistance to Golgi Disrupting Drugs.

Ángela Vázquez-Calvo,Juan-Carlos Saiz,Miguel A Martín-Acebes,Francisco Sobrino,Mónica González-Magaldi,Flavia Caridi

doi:10.3389/fmicb.2016.00612

Abstract

Swine vesicular disease virus (SVDV) is a porcine pathogen and a member of the species Enterovirus B within the Picornaviridae family. Brefeldin A (BFA) is an inhibitor of guanine nucleotide exchange factors of Arf proteins that induces Golgi complex disassembly and alters the cellular secretory pathway. Since BFA has been shown to inhibit the RNA replication of different enteroviruses, including SVDV, we have analyzed the effect of BFA and of golgicide A (GCA), another Golgi disrupting drug, on SVDV multiplication. BFA and GCA similarly inhibited SVDV production. To investigate the molecular basis of the antiviral effect of BFA, SVDV mutants with increased resistance to BFA were isolated. A single amino acid substitution, Q65H, in the non-structural protein 2C was found to be responsible for increased resistance to BFA. These results provide new insight into the relationship of enteroviruses with the components of the secretory pathway and on the role of SVDV 2C protein in this process.

Highlights

Swine vesicular disease virus (SVDV) is a member of the species Enterovirus B within the Picornaviridae family
foot-and-mouth disease virus (FMDV), a picornavirus that is resistant to Brefeldin A (BFA) (Martín-Acebes et al, 2008), and the rhabdovirus vesicular stomatitis virus (VSV), which is highly sensitive to Golgi disrupting agents (Martín-Acebes et al, 2008), were included as controls (Figure 1C)
The BFA-resistant SVDVs here described displayed increased resistance to golgicide A (GCA), supporting that the molecular basis of BFA resistance is related to GBF1, which is inhibited by GCA

Summary

Introduction

Swine vesicular disease virus (SVDV) is a member of the species Enterovirus B within the Picornaviridae family. Positive-strand RNA viruses induce a dramatic remodeling of the intracellular membranes of infected cells aimed to develop an adequate environment for RNA replication (Hsu et al, 2010; Harak and Lohmann, 2015). These structures are considered specialized organelles for viral replication (Hsu et al, 2010; Richards et al, 2014). The proposed mechanism of action of these non-structural proteins involves their interaction with host cell proteins from the secretory pathway, which can vary among picornaviruses (Gazina et al, 2002; Martín-Acebes et al, 2008; van der Linden et al, 2010; Sasaki et al, 2012)

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