Dynamic organization of Herpesvirus glycoproteins on the viral envelope revealed by super-resolution microscopy.

The processes of cell attachment and membrane fusion of Herpes Simplex Virus 1 involve many different envelope glycoproteins. Viral proteins gC and gD bind to cellular receptors. Upon binding, gD activates the gH/gL complex which in turn activates gB to trigger membrane fusion. Thus, these proteins must be located at the point of contact between cellular and viral envelopes to interact and allow fusion. Using super-resolution microscopy, we show that gB, gH/gL and most of gC are distributed evenly round purified virions. In contrast, gD localizes essentially as clusters which are distinct from gB and gH/gL. Upon cell binding, we observe that all glycoproteins, including gD, have a similar ring-like pattern, but the diameter of these rings was significantly smaller than those observed on cell-free viruses. We also observe that contrary to cell-free particles, gD mostly colocalizes with other glycoproteins on cell-bound particles. The differing patterns of localization of gD between cell-free and cell-bound viruses indicates that gD can be reorganized on the viral envelope following either a possible maturation of the viral particle or its adsorption to the cell. This redistribution of glycoproteins upon cell attachment could contribute to initiate the cascade of activations leading to membrane fusion. Author summary The envelopes of Herpesvirus particles contain a variety of different proteins that allow them to infect specific cell types. An essential core set of these proteins is designed to allow viral entry into the cell after adsorption by binding to specific receptors and ultimately inducing fusion between the viral and cellular membranes in a regulated way through a succession of interactions between receptor-binding and fusion-triggering viral proteins. We have identified here for the first time the localization patterns of these essential proteins at the surface of purified virions and we describe how their localization changes after cell attachment. These results illustrate how the dynamics of viral proteins at the surface of the viral particle could participate in optimizing the all-important process of cell binding and membrane fusion.