ND10 are nuclear domains of unknown function that become abundant in response to stress. Infection by herpes simplex virus type 1 (HSV-1) causes the apparent disappearance of these domains, an effect that requires the expression of the immediate early protein ICP0. Previously, we have shown that there are a number of cellular antigens in the ND10. In this report, we show that one of these proteins is PML, a member of the C3HC4 zinc-binding domain family which also includes ICP0. The C3HC4 domain of ICP0 is essential for the apparent release of PML from the ND10, although the interaction of ICP0 with ND10 is determined by a small region near its carboxy terminus. PML and other ND10 proteins are not lost after removal from ND10 but deposited at the nuclear envelope or nuclear envelope modifications during later parts of the replication cycle. ICP0 is required for the onset of low multiplicity infections, and has been implicated in the process of reactivation from HSV latency. Therefore, the interaction between ICP0 and the ND10 domains, specifically PML, may be important for the outcome of virus-cell interactions.