title: The Orf virus E3L homologue is able to complement deletion of the vaccinia virus E3L gene in vitro but not in vivo.
abstract: Orf virus (OV), the prototypic parapoxvirus, is resistant to the effects of interferon (IFN) and this function of OV has been mapped to the OV20.0L gene. The protein product of this gene shares 31% amino acid identity to the E3L-encoded protein of vaccinia virus (VV) that is required for the broad host range and IFN-resistant phenotype of VV in cells in culture and for virulence of the virus in vivo. In this study we investigated whether the distantly related OV E3L homologue could complement the deletion of E3L in VV. The recombinant VV (VV/ORF-E3L) expressing the OV E3L homologue in place of VV E3L was indistinguishable from wt VV in its cell-culture phenotype. But VV/ORF-E3L was over a 1000-fold less pathogenic than wt VV (LD(50) > 5 x 10(6) PFU, compared to LD(50) of wtVV = 4 x 10(3) PFU) following intranasal infection of mice. While wt VV spread to the lungs and brain and replicated to high titers in the brain of infected mice, VV/ORF-E3L could not be detected in the lungs or brain following intranasal infection. VV/ORF-E3L was at least 100,000-fold less pathogenic than wt VV on intracranial injection. Domain swap experiments demonstrate that the difference in pathogenesis maps to the C-terminal domain of these proteins. This domain has been shown to be required for the dsRNA binding function of the VV E3L.
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D016899:Interferon-beta / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009923:Orf virus / Q000235:genetics / Q000472:pathogenicity*; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014615:Vaccinia / Q000503:physiopathology / Q000821:virology; D014616:Vaccinia virus / Q000235:genetics / Q000472:pathogenicity*; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
chemical_list: C081374:E3L protein, Vaccinia virus; C117524:E3L protein, orf virus; C112834:OV20.0L protein, Orf virus; D016601:RNA-Binding Proteins; D014764:Viral Proteins; D016899:Interferon-beta