Analysis of the glycoproteins of Seoul orthohantavirus strain B1 associated with fusion activity

We analyzed two virus variants (S1 and L1) from Seoul orthohantavirus strain B1. Strain B1 produces large opaque plaques when plated on Vero E6 cell monolayers. However, although the L1 variant produced the large opaque plaques common to the strain, the variant S1 produced small clear ones on Vero E6 cells. Five days after Vero E6 cells were infected with the S1 variant, polykaryons formed spontaneously. However, the cells infected with the L1 variant did not show the formation of syncytia. An analysis of the pH dependency of the cell fusion demonstrated that the L1 variant could induce cell fusion, but only at a pH that was 0.2 units lower than the pH at which the S1 variant induced it. Sequencing of the M genome segment of the two virus variants revealed amino acid substitutions at 4 positions in the Gn and Gc gene products of the S1 variant. Two of these substitutions occurred in the extracellular domain of Gn and changed the charge of the protein. Our findings suggest that these amino acid substitutions caused the S1 variant Gn protein to induce fusion at an elevated pH.