Fig. 3: Newly identified conserved motif I in sNSV polymerases.
a, Hydrogen bond (dashed yellow line) between Arg1289 of ordered fingertips and Lys1008 of the core lobe in the TSWV polymerase active site. b, The conservation of lysine (motif I) is shown in an alignment in bunyaviruses (TSWV (Orthotospovirus); BADUV, Badu virus (Phasivirus); EMARaV, European mountain ash ringspot-associated virus (Emaravirus); FERV, Ferak virus (Orthoferavirus); GOLV, Gouleako virus (Goukovirus); HEBV, Herbert virus (Herbevirus); HTNV (Orthohnatavirus); JONV, Jonchet virus (Orthojonvirus); KIGV, Kigluaik phantom virus (Orthophasmavirus); LACV (Orthobunyavirus); LASV (Mammarenavirus); MACV (Mammarenavirus); RSV, rice stripe virus (Tenuivirus); RVFV (Phlebovirus); SFTSV (Phlebovirus); SNV (Orthohnatavirus)) and orthomyxoviruses (FluA). The blue triangle highlights the position of the conserved lysine. c–j, The surroundings of motif I are illustrated for HTNV (c), LACV (d), LASV (e), MACV (f), RVFV (g), SFTSV (h), SNV (i) and FluA (j) with the neighbouring motifs A–H. k, N. benthamiana leaves were inoculated with wild-type TSWV infectious clone or its mutant via agro-infiltration. Accumulation of eGFP fluorescence expressed from rescued TSWV was detected in N. benthamiana leaves at 60 h post inoculation. Scale bars, 200 μm (left). Expression levels of eGFP, L and its mutant were analysed via western blot (top right), and the S genomic RNA (red arrowhead) and eGFP mRNA transcripts (green arrowhead) were analysed via northern blot using DIG-labelled antisense eGFP probes (bottom right). Ethidium bromide staining of ribosomal RNA (rRNA) was used as the RNA loading control.
