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Human herpesvirus 6 (HHV-6) is a lymphotropic herpesvirus, and in vitro, it can productively infect many of the same cell types that human immunodeficiency virus (HIV) infects. Simultaneous infection of T cells by HIV and HHV-6 can lead to both activation of the HIV promoter and acceleration of the cytopathic effects. Several HHV-6 genes have been demonstrated to activate HIV promoter expression. Among them is a cDNA clone, pCD41 (U27), which codes for the HHV-6 DNA polymerase accessory protein. We have now further characterized the transcription pattern in the pCD41 locus and identified at least six RNA species, ranging in size from 1.2 to 4.5 kb. Northern (RNA) blot analyses showed no significant difference in RNA patterns between the HHV-6 variant A (GS) and variant B (Z29) viruses. All the RNA species detected by pCD41 are polyadenylated and polyribosome associated, suggesting that they may be actively engaged in protein synthesis. Cycloheximide and phosphonoacetic acid inhibition assay results indicate that all the pCD41 RNA species belong to the herpesviral early-late family. Using primer extension and S1 mapping techniques, the 5’ and 3’ ends of each transcript were mapped to different positions, and no splicing was observed.