title: Variola virus E3L Zα domain, but not its Z-DNA binding activity, is required for PKR inhibition.
abstract: Responding to viral infection, the interferon-induced, double-stranded RNA (dsRNA)-activated protein kinase PKR phosphorylates translation initiation factor eIF2α to inhibit cellular and viral protein synthesis. To overcome this host defense mechanism, many poxviruses express the protein E3L, containing an N-terminal Z-DNA binding (Zα) domain and a C-terminal dsRNA-binding domain (dsRBD). While E3L is thought to inhibit PKR activation by sequestering dsRNA activators and by directly binding the kinase, the role of the Zα domain in PKR inhibition remains unclear. Here, we show that the E3L Zα domain is required to suppress the growth-inhibitory properties associated with expression of human PKR in yeast, to inhibit PKR kinase activity in vitro, and to reverse the inhibitory effects of PKR on reporter gene expression in mammalian cells treated with dsRNA. Whereas previous studies revealed that the Z-DNA binding activity of E3L is critical for viral pathogenesis, we identified point mutations in E3L that functionally uncouple Z-DNA binding and PKR inhibition. Thus, our studies reveal a molecular distinction between the nucleic acid binding and PKR inhibitory functions of the E3L Zα domain, and they support the notion that E3L contributes to viral pathogenesis by targeting PKR and other components of the cellular anti-viral defense pathway.
authors: Meghna
authors: M
authors: Thakur
authors: Eun Joo
authors: EJ
authors: Seo
authors: Thomas E
authors: TE
authors: Dever
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D043542:DNA, Z-Form / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D012441:Saccharomyces cerevisiae; D012901:Variola virus / Q000502:physiology*; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics; D019892:eIF-2 Kinase / Q000037:antagonists & inhibitors / Q000737:chemistry*
chemical_list: D043542:DNA, Z-Form; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D014764:Viral Proteins; D019892:eIF-2 Kinase
keywords: E3L; PKR; eIF2; translation