Multiple adjacent or overlapping loci affecting the level of gC and cell fusion mapped by intratypic recombinants of HSV-1☆

Abstract We have prepared and analyzed 40 HSV-1 intratypic recombinants with regard to plaque morphology and glycoprotein C(gQ phenotypes. Vero cells have been cotransfected with the intact genome of HSV-1(F) and cloned or uncloned DNA fragments from HSV-1(MP) and recombinants inducing the fusion of Vero cells [syncytial (Syn) recombinants] have been selected and purified. Marker transfer of the Syn phenotype has been observed with the cloned Bam HIL and B fragments (0.706 – 0.745 and 0.745 – 0.810 map units, respectively) as well as with the uncloned Hpa I TXO fragment (0.710 – 0.761) from MP DNA. No marker transfer has been observed with F DNA alone or with the cloned BamHI N fragment (0.863 – 0.898 map units). When viruses expressing the Syn phenotype in Vero cells were tested in HEp-2 cells, three kinds of recombinants were observed. Members of the first class expressed a wild type, cytoaggregating (Syn + ), plaque morphology in these cells. Members of the second class induced the complete fusion (Syn phenotype) of the cells. Members of the third class induced an intermediate plaque morphology, characterized by the formation of groups of polykaryocytes (fused cells) but without formation of a complete syncytium. All recombinants expressing the Syn+ phenotype in HEp-2 cells were also gC + , whereas recombinants expressing the Syn phenotype in these cells were gC − with one exception, in which low levels of gC could be detected (but clearly less than with HSV-1(F)). Concerning polykaryocytic class of recombinants, some of them were gC + while others expressed only low amounts of gC; no gC − virus was observed within this class of recombinants. The three classes of recombinants were observed with each of the cloned Bam HI L and B fragments and also with the Hpa I TXO fragment, suggesting the existence of multiple adjacent or overlapping loci affecting plaque morphology and the control of the accumulation or the synthesis of gC at both sides of 0.745 map units.