We have examined the nuclear and cytoplasmic distribution of the latency-associated transcripts (LATs) of HSV-1. During latency these transcripts accumulate in the nuclei of neurons in the peripheral and central nervous system of infected animals. However, our Northern blot analyses demonstrate that the 2-kb LAT is found in the cytoplasm of HSV-1-infected CV-1 cells, and brainstems of HSV-1 productively infected mice. Like the nuclear LAT from latently infected tissue, most of the cytoplasmic 2-kb LAT from lytically infected CV-1 cells is unpolyadenylated. In order to determine if cytoplasmic LAT could be translated, we compared its distribution with that of glycoprotein C mRNA in polysome profiles from HSV-1-infected tissue culture cells. specific association of RNAs to polysomes was verified by disruption of polysomes with EDTA or puromycin. Analyses of numerous experiments indicate that most of the cytoplasmic 2-kb LAT migrates at the position of ribosomal subunits in polysome profiles. Thus, the 2-kb LAT may not be efficiently translated during productive infection. This suggests that if the 2-kb LAT is indeed translated, its translation may be tightly regulated during HSV-1 infection, possibly in a cell type- or cell cycle-specific manner. Another possibility is that the 2-kb LAT is not a translated RNA but may have another function, possibly related to translation as indicated by its apparent association to ribosomal complexes. (C) 1994 Academic Press, Inc.