Attenuation of Rabies Virus Replication and Virulence by Picornavirus Internal Ribosome Entry Site Elements

Max von Pettenkofer-Institute and Gene Center, Ludwig-Maximilians-University, Munich, Germany.
Journal of Virology (Impact Factor: 4.44). 02/2009; 83(4):1911-9. DOI: 10.1128/JVI.02055-08
Source: PubMed


Gene expression of nonsegmented negative-strand RNA viruses is regulated at the transcriptional level and relies on the canonical
5′-end-dependent translation of capped viral mRNAs. Here, we have used internal ribosome entry sites (IRES) from picornaviruses
to control the expression level of the phosphoprotein P of the neurotropic rabies virus (RV; Rhabdoviridae), which is critically required for both viral replication and escape from the host interferon response. In a dual luciferase
reporter RV, the IRES elements of poliovirus (PV) and human rhinovirus type 2 (HRV2) were active in a variety of cell lines
from different host species. While a generally lower activity of the HRV2 IRES was apparent compared to the PV IRES, specific
deficits of the HRV2 IRES in neuronal cell lines were not observed. Recombinant RVs expressing P exclusively from a bicistronic
nucleoprotein (N)-IRES-P mRNA showed IRES-specific reduction of replication in cell culture and in neurons of organotypic
brain slice cultures, an increased activation of the beta interferon (IFN-β) promoter, and increased sensitivity to IFN. Intracerebral
infection revealed a complete loss of virulence of both PV- and HRV2 IRES-controlled RV for wild-type mice and for transgenic
mice lacking a functional IFN-α receptor (IFNAR−/−). The virulence of HRV2 IRES-controlled RV was most severely attenuated and could be demonstrated only in newborn IFNAR−/− mice. Translational control of individual genes is a promising strategy to attenuate replication and virulence of live nonsegmented
negative-strand RNA viruses and vectors and to study the function of IRES elements in detail.

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Available from: Stefan Finke, Oct 01, 2014
    • "In vivo studies demonstrated that deletion of the dynein LC8 binding domain retained pathogenicity in suckling mice (Mebatsion 2001) but was nonpathogenic in adult mice (Tan et al. 2007). In another study, the internal ribosome entry site (IRES) from picornaviruses was introduced to control the expression of P, and an in vivo study showed reduction of replication in cell culture, increased activation of the beta interferon (IFN-β) promoter, and attenuation in mice infected intracerebrally (Marschalek et al. 2009). Taken together, targeting the P protein through either mutating the dynein LC8 binding domain or translational control of expression is a promising strategy to attenuate virus virulence of live RABV. "
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    • "For example, removal of Dynein Light Chain 8 binding site motif substantially reduced viral transcription and replication in the central nervous system [32]. Another strategy was to make the expression of the essential phosphoprotein dependent on translation and not transcription [33]. The SAD dIND1 construct uses a different approach which is aimed at inducing an improved innate immune response in vaccinated animals. "
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