MicroRNA-134 regulates poliovirus replication by IRES targeting

Abhijeet A Bakre,Byoung-Shik Shim,Ralph A Tripp

doi:10.1038/s41598-017-12860-z

Abhijeet A Bakre, Byoung-Shik Shim + Show 1 more

Open Access

https://doi.org/10.1038/s41598-017-12860-z

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Abstract

Global poliovirus eradication efforts include high vaccination coverage with live oral polio vaccine (OPV), surveillance for acute flaccid paralysis, and OPV “mop-up” campaigns. An important objective involves host-directed strategies to reduce PV replication to diminish viral shedding in OPV recipients. In this study, we show that microRNA-134-5p (miR-134) can regulate Sabin-1 replication but not Sabin-2 or Sabin-3 via direct interaction with the PV 5′UTR. Hypochromicity data showed miR-134 binding to Sabin-1 and 3 but not Sabin-2 IRES. Transfection of a miR-134 mimic repressed translation of Sabin-1 5′UTR driven luciferase validating the mechanism of miR-134-mediated repression of Sabin-1. Further, site directed mutagenesis of the miR-134 binding site in Sabin-1 IRES relieved miR-134-mediated repression indicating that these regulatory molecules have an important role in regulating the host gene response to PV. Binding of miR-134 to Sabin-1 IRES caused degradation of the IRES transcript in a miR-134 and sequence specific manner. The miR-134 binding site was found to be highly conserved in wild type PV-1 as well as EV71 strains indicating that miR-134 may regulate function of these IRES sequences in circulation.

Highlights

Global poliovirus eradication efforts include high vaccination coverage with live oral polio vaccine (OPV), surveillance for acute flaccid paralysis, and OPV “mop-up” campaigns
We recently showed that upregulation of miR-134 during PV infection causes a reduction in replication for both Sabin-1, strains but is absent in PV-2 (Sabin-2) and Sabin-3, as well as Enterovirus 71 (EV71)[30,31]
We demonstrated that miR-134 downregulation of Ras-related nuclear protein (RAN) inhibits both PV and EV71 replication

Summary

Introduction

Global poliovirus eradication efforts include high vaccination coverage with live oral polio vaccine (OPV), surveillance for acute flaccid paralysis, and OPV “mop-up” campaigns. Site directed mutagenesis of the miR-134 binding site in Sabin-1 IRES relieved miR-134-mediated repression indicating that these regulatory molecules have an important role in regulating the host gene response to PV. The miR-134 binding site was found to be highly conserved in wild type PV-1 as well as EV71 strains indicating that miR-134 may regulate function of these IRES sequences in circulation. Though OPV and IPV confer lifelong protection, OPV vaccinees can shed and contribute to vaccine-derived PV (VDPV), immune deficiency-associated PV (iVDPV) or circulating vaccine-derived poliovirus (cVDPV) causing concerns of reversion to wild type. Computational and experimental evidence suggest that up to 60% of the eukaryotic transcriptome is under miRNA regulation[17], and this class of regulatory molecules has important roles in determining host gene response to infection as well[16]. Increasing miR-555 activity inhibits a host factor critical for PV replication, heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNP C)[29], which inhibits PV replication[28]

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