The ionophore monensin inhibited the formation of herpes simplex virus type 2 (HSV-2) particles by about 30% but the yields of infectious particles were reduced to 5% and 1% for cell-associated and extracellular virus, respectively. The presence of monensin did not affect the processing of the two viral glycoproteins gB-2 and gG-2. However, two other glycoproteins, gC-2 and gD-2, were not processed to their fully mature forms in the monensin-treated cells and only the faster moving pgC-2 and pgD-2 were detected. The cell-associated virus particles contained the glycoproteins gB-2, gC-2, gD-2 and gG-2, whereas the extracellular virus particles contained only gG-2 glycoprotein. These results suggest that HSV-2 particles containing all the viral glycoproteins are transported via the Golgi apparatus to the cell surface but that virus particles containing only gG-2 may follow a different pathway for transport and release.