Attenuation of Rabies Virulence: Takeover by the Cytoplasmic Domain of Its Envelope Protein

Institut Pasteur, 75724 Paris, France.
Science Signaling (Impact Factor: 6.28). 01/2010; 3(105):ra5. DOI: 10.1126/scisignal.2000510
Source: PubMed


The capacity of a rabies virus to promote neuronal survival (a signature of virulence) or death (a marker of attenuation) depends on the cellular partners recruited by the PDZ-binding site (PDZ-BS) of its envelope glycoprotein (G). Neuronal survival requires the selective association of the PDZ-BS of G with the PDZ domains of two closely related serine-threonine kinases, MAST1 and MAST2. Here, we found that a single amino acid change in the PDZ-BS triggered the apoptotic death of infected neurons and enabled G to interact with additional PDZ partners, in particular the tyrosine phosphatase PTPN4. Knockdown of PTPN4 abrogated virus-mediated apoptosis. Thus, we propose that attenuation of rabies virus requires expansion of the set of host PDZ proteins with which G interacts, which interferes with the finely tuned homeostasis required for survival of the infected neuron.

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Available from: Muriel Delepierre, Oct 07, 2015
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    • "To generate SADcvsG, the cytoplasmic domain of SADG was replaced by that of CVSG. The cytoplasmic domain of CVSG, rather than that of HEPG, was selected since the cytoplasmic domain of RVGs of attenuated strains, such as HEP-Flury, might induce cell apoptosis (Prehaud et al., 2010). PCR was utilized to replace the cytoplasmic domain of EnvARGCD glycoprotein, which originates from the SAD strain, with that of the CVS strain. "
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    ABSTRACT: Rabies virus (RV) has been widely used to trace multi-synaptic neuronal circuits. The recent development of glycoprotein-deficient rabies virus (RV-ΔG) expressing various proteins has enabled analyzes of both the structure and function of neuronal circuits. The main advantage of RV-ΔG is its ability to trace monosynaptic circuits by the complementation of rabies virus glycoprotein (RVG), but it has the disadvantage of cytotoxicity. Several strain variants of RV have different biological characteristics, such as synaptic spreading and cytotoxicity, mainly due to amino acid mutations in RVG. We developed an improved protocol for the production of a highly attenuated strain of RV-ΔG and assessed whether RVG variants affect rabies monosynaptic tracing and the health of infected neurons. We demonstrated that (1) rabies monosynaptic tracing with RVG variants traced different subsets of presynaptic partners, (2) RVG of the attenuated strain also labeled astrocytes, and (3) the cytotoxicity of RV-ΔG did not depend on RVG but on RV-ΔG. These findings indicate that RVG variants are an important determinant of rabies monosynaptic tracing.
    Frontiers in Neuroanatomy 01/2014; 7:47. DOI:10.3389/fnana.2013.00047 · 3.54 Impact Factor
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    • "Construction of recombinant cDNA clones. The SN strain of RABV was derived from the SAD B19 cDNA clone as described previously (Faber et al., 2004; McGettigan et al., 2001; Morimoto et al., 2000; Pulmanausahakul et al., 2008). Glycoproteins (Gs) from EBLV- 1 and EBLV-2 were amplified using Elongase (Invitrogen) and the following primers: EBLV1GEcoRVF (59-TCAGATATCATGTTAC- TCTCTAC-39) and EBLV1GNheIR (59-AAGGCTAGCTTATGACT- CACC-39); EBLV2GEcoRVF (59-GATCTCGATATCATGCCATTCC- 39) and EBLV2GNheIR (59-AGCAAGGCTAGCTTAAGACTG-39). "
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    ABSTRACT: European bat lyssaviruses type-1 (EBLV-1) and -2 (EBLV-2) circulate within bat populations throughout Europe and are capable of causing disease indistinguishable from that caused by classical rabies virus (RABV). However, the determinants of viral fitness and pathogenicity are poorly understood. Full-length genome clones based on the highly attenuated, non-neuroinvasive, RABV vaccine strain (SAD-B19) were constructed with the glycoprotein (G) of either SAD-B19 (SN), of EBLV-1 (SN-1) or EBLV-2 (SN-2). In vitro characterization of SN-1 and SN-2 in comparison to wildtype EBLVs demonstrated that the substitution of G affected the final viral titer, and antigenicity. In vivo, following peripheral infection with a high viral dose (104 ffu), animals infected with SN-1 had reduced survivorship relative to infection with SN, resulting in similar survivorship to EBLV-1. The histopathological changes and antigen distribution observed for SN-1 were more representative of those observed with SN than with EBLV-1. EBLV-2 was unable to achieve a titer equivalent to that of the other viruses. Therefore, a reduced dose experiment (103 ffu) was undertaken in vivo to compare EBLV-2 and SN-2, which resulted in 100% survivorship for all recombinant viruses (SN, SN-1 and SN-2) whilst clinical disease developed in mice infected with the EBLVs. These data indicate that interspecies replacement of G has an effect on viral titer in vitro, probably as a result of sub-optimal G:M interactions, and influences the survival outcome following a peripheral challenge with a high viral titer in mice.
    Journal of General Virology 10/2012; 94(Pt 2). DOI:10.1099/vir.0.048827-0 · 3.18 Impact Factor
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    • "They include the single surface glycoprotein (G) gene, which in RABV is a major factor determining neurotropism and neuroinvasiveness. Specifically, the G protein is responsible for attachment to receptors on neuronal cells (Lafon, 2005), retrograde transport of viral particles (Klingen et al., 2007), transsynaptic spread (Wickersham et al., 2007b; Etessami et al., 2000) and preservation of neuronal integrity (Prehaud et al., 2010), which all is required for gaining access to the CNS, where the viruses multiply extensively (Dietzschold et al., 2008). In addition , species-specific factors associating with the viral RNA synthesis machinery (which comprises the viral N, P, M, and L proteins) may limit the efficiency of replication in a heterologous host (Finke and Conzelmann, 2005; Finke and Conzelmann, 2003). "
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    ABSTRACT: Rabies is a zoonosis still claiming more than 50 000 human deaths per year. Typically, human cases are due to infection with rabies virus, the prototype of the Lyssavirus genus, but sporadic cases of rabies-like encephalitis caused by other lyssaviruses have been reported. In contrast to rabies virus, which has an extremely broad host range including many terrestrial warm-blooded animals, rabies-related viruses are associated predominantly with bats and rarely infect terrestrial species. In spite of a very close genetic relationship of rabies and rabies-related viruses, the factors determining the limited host range of rabies-related viruses are not clear. In the past years the importance of viral countermeasures against the host type I interferon system for establishment of an infection became evident. The rabies virus phosphoprotein (P) has emerged as a critical factor required for paralysing the signalling cascades leading to transcriptional activation of interferon genes as well as interferon signalling pathways, thereby limiting expression of antiviral and immune stimulatory genes. Comparative studies would be of interest in order to determine whether differential abilities of the lyssavirus P proteins contribute to the restricted host range of lyssaviruses.
    Berliner und Münchener tierärztliche Wochenschrift 05/2012; 125(5-6):209-18. · 0.82 Impact Factor
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