Attenuation of rabies virulence: Takeover by the cytoplasmic domain of its envelope protein

Institut Pasteur, 75724 Paris, France.
Science Signaling (Impact Factor: 7.65). 01/2010; 3(105):ra5. DOI: 10.1126/scisignal.2000510
Source: PubMed

ABSTRACT 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, Aug 29, 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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    • "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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    • "Apart from peptide residues at p0 and pÀ2, which are the hallmarks for recognition by PDZs, the role of other residues of the PBM is more variable . Some studies suggested that the contribution of residues at pÀ1 and pÀ3 to the overall binding event is minor whereas in other studies PDZ domains exhibited clear preferences for certain residues at these sites over others [39] [40] [41] [42] [43]. More and more studies now converge on the idea that PBMs should at least be extended to pÀ4 as residues at this position have also been observed to significantly contribute to the binding to PDZs [19] [35]. "
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