Fig. 4: Broad-spectrum inhibitory effect of SAA on arenaviral CENs.
a Phylogenetic tree of the pathogens from family Phenuiviridae, Nairoviridae, Arenaviridae, Hantaviridae, and Peribunyaviridae in the order of Bunyavirales were made based on L protein sequences, pathogens mentioned in this study were marked. b Alignment of the primary amino acid sequences and secondary spatial structures of LCMV, MACV, and LASV CENs, and analysis on enzyme active pocket. Blue triangles indicate the active sites of LCMV CEN. Red triangles indicate mutational sites of LCMV CEN. Amino acid sites of PD-D/E(X)K are marked by red pentagrams. c The inhibitory effect of SAA on the cleavage of MACV and LASV CENs on ssRNA substrate was examined at a number of different time points via acrylamide-urea gel electrophoresis. Gray analysis of the corresponding strips was calculated into the inhibition rate, shown in the right panel. The divalent metal ion chelator, EDTA, was used as a positive control. The representative image was derived from three replicate experiments. d IC50 of SAA on MACV and LASV CENs were determined using FRET. BXA was used as a control. e SAA inhibited MACV and LASV CENs in a substrate-competing manner. f Affinity of SAA to MACV and LASV CENs was detected using SPR. Data were from once representative experiment. g In vitro inhibitory activity of SAA against CHAPV CEN, DANV CEN, GTOV CEN, JUNV CEN, LUJV CEN and SABV CEN in Arenaviridae. BXA was used as a control. Data from enzymatic assays were obtained from at least three independent experiments.
