On the basis of predicted amino acid sequence characteristics, herpes simplex virus type 1 gene UL10 is thought likely to encode a membrane protein with eight potential transmembrane regions. Previously, a protein with an apparent M(r) 47,000 on SDS-PAGE was identified as a product of this gene. Here we have further characterized this protein, and show that it is modified by N-linked glycosylation, associates with membranes from infected cells, and is a component of the virus particle. It is not essential for virus growth in tissue culture. To investigate its role in vivo a deletion mutant lacking the majority of the UL10 open reading frame was constructed (UL10-del). The in vitro growth properties of this virus were consistent with previous studies; it grew to give slightly lower yields than wild-type and revertant viruses, and had no apparent temperature-sensitive or host range phenotype. In vivo, in a mouse model, UL10-del was capable of establishing a latent infection, although it was impaired for growth at the periphery, and for spread to and/or growth within the nervous system relative to wild-type or revertant viruses.