Syncytial mutations of herpes simplex virus type 1 (HSV-1) strains ANG, ANG path, HFEM, tsB5 and HSZP cause extensive cell fusion and were mapped to the cytoplasmic domain of glycoprotein B (gB), within the syn 3 locus. These strains are so far the only ones which show the phenotype ‘fusion from without’ (FFWO): 60 min after infection with high m.o.i., cells in a tissue culture are fused without transcription and translation of the viral genome. In this report we detected, using the recombinants 27/III and K-7, that an amino acid exchange from Ala to Val at aa position 854 of gB is the main determinant for FFWO activity of strains ANG, ANG path and recombinant K-7. The transfer of this mutation to wild-type strains KOS and 17 syn by co-transfection results in recombinants KOS-854Q, 17-syn3, 17-syn3a and 17-syn3b. As a selection marker we used the cyclosporin A resistance of fusion which was shown to be a unique characteristic of syn 3 locus mutants. The recombinants show the FFWO phenotype in BHK cells but not in Vero cells. FFWO was shown to be cell-type dependent by comparing the number of p.f.u. needed to induce FFWO in various cell types.


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